Central Composite Design for Optimizing the Biogenic fabrication of Platinum Nanoparticles Using bacillus cereus and TheirAntimicrobial, Antioxidant, and catalytic Potential

Central Composite Design for Optimizing the Biogenic fabrication of Platinum Nanoparticles Using bacillus cereus and TheirAntimicrobial, Antioxidant, and catalytic Potential | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Central Composite Design for Optimizing the Biogenic fabrication of Platinum Nanoparticles Using bacillus cereus and TheirAntimicrobial, Antioxidant, and catalytic Potential Saba Ibrahim, Zahoor Ahmad, Muhammad Mujahid This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6922293/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract To progress one step biosynthetic process, reciprocal effect of reaction variables like concentration of platinum salt solution, inoculum size, temperature, time, and pH was considered by making use of central composite design of response surface methodology. The optimized set of reaction conditions were platinum salt solution (1mmol) = 3.0ml, inoculum size ( bacillus cereus ) = 3ml, temperature = 43.2 ◦ C, time = 58.4hours, and pH = 8.0. The biogenic platinum nanoparticles were categorized by UV-vis spectroscopy, FTIR, DLS-PSA, XRD and SEM. The UV-visible spectra showed a surface plasmon response at 290nm. FTIR spectra revealed the existence of enzymatic proteins responsible for fabrication and stabilization of platinum nanoparticles. DLS-PSA, XRD and SEM confirmed the formation of isotropic and spherical platinum nanoparticles of size 6.29 to 25.0 nm with a specific surface area of 194.95m 2 /g. Moreover, gram positive bacterial strains Staphylococcus epidermidis, Staphylococcus aureus, Bacillus subtilis, Bacillus licheniformis and gram-negative bacterial strains Escherichia coli, Salmonella enteria, Porteus mirabilis, Pseudomonas aeruginosa were tested for antibacterial activity by employing disc diffusion and broth macro dilution methods. DPPH (2,2-diphenyl-1-picrylhydrazyl) assay was carried out to monitor the free radical scavenging ability of biogenic platinum nanoparticles. Furthermore, the degradation rate of methyl orange and methylene blue in presence pf platinum nanoparticles as catalyst was investigated successfully. The bio fabricated platinum nanoparticles found applications in biomedical and environmental remediation. Platinum nanoparticles Bacillus Cereus RSM CCD Antimicrobial activity Antioxidant Catalytic potential Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 INTRODUCTION Microbial synthesis of metal nanoparticles is of colossal concern to modern nanotechnology as it is simple, cost-effective, and eco-friendly. In the present study, a novel green method for biogenic synthesis of platinum nanoparticles has been developed using a gram-positive bacterial strain [ 1 ]. Green synthesis of metal nanoparticles is attaining extraordinary consideration in various applications [ 2 ] [ 3 ]. To synthesize metal nanosized particles, various methods have been used, including colloidal systems and chemical reduction [ 4 ]. These approaches are favored in research due to their simplicity, reliability, and strong potential for biomedical applications, while maintaining the structural integrity of the nanoparticles [ 5 ]. Furthermore, the preparation of nanoparticles from transition metals has a diversity of functions including material science, chemistry, and medicine[ 6 ]. Transition metal nanoparticles are vital due to their biocompatibility and eco-friendly versatile nature. Nanoparticles of Pb, Cu, Pd, Ag, Au, and Pt find applications in catalysis, pollution remediation, drug delivery, wastewater [ 7 ]. The choice of method for synthesis of metal nanoparticles is very important as during synthesis proceedings like kinetic interaction of the metal with reducing agent, retention of the stabilizing agent with metal nanoparticles and distinct experimental approaches causes a robust effect on constancy, morphology, physical and chemical properties [ 8 , 9 ]. Among these metal nanoparticles, the synthesis of platinum nanoparticles, in particular, their biogenic fabrication has attracted great interest as it is sustainable, green, and cost-effective. Platinum nanoparticles have remarkable applications due to their large surface area, corrosion, and chemical resistance. Platinum nanoparticles are used as dental material ingredient [ 10 ], coating of medical devices [ 11 ], water treatment [ 12 ], cancer treatment [ 13 ], antiaging [ 14 ], cosmetics [ 15 ], dye degradation [ 16 ], and much more. Generally, two systems are used for the biogenic fabrication of metal nanoparticles. First is the use of different parts of plants like roots, stems, leaf, fruit and flower [ 17 ], [ 18 ]. Plants have rich amounts of secondary metabolites which are responsible for reduction of metal ion to metal nanoparticles. However, concentration of these secondary metabolites is different in different parts of plants and methods of extraction influence their concentration [ 19 ] so the quality and quantity of nanoparticles get compromised. Second is microbial system which includes bacteria, fungi, algae, actinomycetes, yeast and are more rapid. Microorganisms have the ability to produce metal nanoparticles intracellularly and extracellularly. In both, reductase enzyme is responsible for conversion of metal ion into nanoparticles [ 3 ]. However, microbial systems have been less focused for synthesis of platinum nanoparticles. In the present work, a novel microbial synthesis of platinum nanoparticles through bacillus cereus has been reported. As conditions of an experiment such as pH, temperature, the concentration of metal salt, time of reaction, inoculum size of bacillus cereus , play a vital role in the biosynthesis of platinum nanoparticles. A manual set up to study one aspect at a time by making others persistent for these experimental conditions is long, tedious, time-consuming, costly, and causes errors so it is avoided. Response surface methodology (RSM) is a blend of mathematical and statistical procedures used for designing experiments, construction of models, assessing the operative factors, and most significant is searching for the optimal conditions for a required response [ 20 ]. This has to be done by considering three main stepladders as a performance of statistically planned experiments, approximating the coefficient in the mathematical model, and forecasting the response by checking the suitability of the model [ 21 ]. We used central composite design (CCD) of response surface methodology (RSM) which is mostly adapted for optimization of multiple factors varying simultaneously. It can provide comprehensive intuition of direct, curved, and pair-wise mutual effects of variables on response. The central composite design is a more efficient, available operative tool for the optimization of complex processes [ 22 ]. Few reports are available regarding the synthesis of platinum nanoparticles through microbes[ 23 ], [ 24 ]. However, no report is available showing RSM mediated optimization of biosynthesis of platinum nanoparticles through bacillus cereus . In the current study, bacillus cereus has been used to mediate the biosynthesis of platinum nanoparticles through the reduction of ionic platinum into metallic nano platinum. The procedure of biosynthesis was optimized via the central composite design technique in ordered to get the proper biosynthetic conditions and understanding their interactive effects. Platinum nanoparticles were characterized through simple systematic techniques of UV-visible spectrophotometer, scanning electron microscopy (SEM), Furrier transform infra-red (FTIR) spectroscopy, Dynamic light scattering-particle size analyzer (DLS-PSA), and X-ray diffraction (XRD). The present study is planned to regulate methods for bacterial synthesis of small-sized, stable, and mono-dispersed platinum nanoparticles having biomedical and environmental applications. RESULTS AND DISCUSSION In the present work, central composite design (CCD) of response surface methodology (RSM) was employed to find the optimized experimental conditions of (A) metal salt concentration (ml of 1mM), (B) inoculum size in ml ( bacillus cereus ), (C) temperature, (D) time, and (E) pH for green synthesis of platinum nanoparticles. Response Surface Methodology Design-Expert-12 software was employed for the purpose that can be accessed by the link https://www.statease.com/software/design-expert/ . The central composite design gave a model of twenty-six experiments and the response of each experiment was represented as absorbance at 290nm (Table 1 ). The design was used to attain the best surface plasmon response of synthesized platinum nanoparticles with reciprocal peak intensity. Basically, the process of optimization through CCD of RSM can be achieved by performing statistically designed experiments, then assessing the design coefficients, and finally examining the design fitness [ 25 ]. After performing all the twenty-six experiments, results in the form of coded factors were presented by a polynomial quadratic equation. SPR Intensity = + 3.94 + 0.0659 A + 0.0824 B + 0.7797 C − 0.0769 D -0.0110 E + 0.3582 AB – 0.7566 AC + 0.0050 AD + 0.1016 AE – 0.0956 BC + 0.5710 BD + 0.6551 BE – 0.2714 CD – 0.2648 CE + 0.6368 DE – 0.0429 A 2 – 0.0790 B 2 – 0.4348 C 2 – 0.1092 D 2 – 0.7665 E 2 . This coded factor equation helps to make estimates about the retort for given stages of each factor. The symbiotic and combative effects of these factors on the feedback were represented with positive and negative symbols. The higher levels of the factors are coded as + 1 and lower levels are coded as -1. The coded equation is beneficial in the identification of the relative effect of the factors by equating the factor coefficients. Based on this equation, the parity plot explains the proper correlation between actual and forecast responses. The difference between actual and predicted response is proportional to the space of these values from the slanted line (Fig. 1a). The level of significance of each factor in the regression model in terms of linear, quadratic, and interactive factors, was determined by p-values. The integrity of the present model was checked by the determinant coefficient (R 2 ) which justifies the fluctuation in the predicted and actual response. The R 2 of 0.9908 exhibits that 99.08% variation in platinum nanoparticle synthesis is because of independent variables. The model R 2 is in close agreement with adjusted R 2 which is 0.9539. For a good model, R 2 should be in the range of 0–1[ 26 ]. Adequate precision means a signal to noise ratio. A value bigger than 4 is needed. The value of adequate precision 15.4515 stated satisfactory signals. The f-value of 5.14 and p-value of 0.0861 of non-significant lack of fit recommend that the adopted regression model is significant. The outcomes of the analysis of variance (ANOVA) are fundamental to evaluate the implication, accuracy, and authenticity of the statistical model and are given in (Table 2 ). The model f-value 26.87 and model p-value 0.0009 suggest that the model and model terms C, AB, AC, BD, BE, DE, C 2 , E 2 are highly recognizable. A comparatively lower value of CV (variation coefficient) 6.96% exhibits the efficiency, credibility, and accuracy of model trials. Table 1 The central composite design (CCD) of the experiments for the biosynthesis of platinum nanoparticles through bacillus cereus . Factor 1 Factor 2 Factor 3 Factor 4 Factor 5 Response Std Run A: Salt (1mmol) B: Inoculum Size C: Temperature D: time E: pH Absorbance mL mL ◦ C Hours Mol/L of H + ions 290nm 24 1 5.5 5.5 35 42 7 3.37 25 2 5.5 5.5 35 42 7 3.61 10 3 3.02905 7.97095 43.2365 58.473 5.9018 3.25 6 4 7.97095 3.02905 26.7635 58.473 8.0982 1.40 3 5 3.02905 7.97095 43.2365 25.527 8.0982 3.25 2 6 7.97095 3.02905 43.2365 58.473 5.9018 1.75 23 7 5.5 5.5 35 42 7 3.80 14 8 5.5 1 35 42 7 4.00 15 9 5.5 10 35 42 7 3.96 13 10 10 5.5 35 42 7 3.80 9 11 7.97095 3.02905 43.2365 25.527 8.0982 2.33 8 12 3.02905 7.97095 26.7635 58.473 8.0982 2.37 5 13 7.97095 7.97095 26.7635 25.527 8.0982 2.57 26 14 5.5 5.5 35 42 7 3.71 20 15 5.5 5.5 35 42 5 1.56 19 16 5.5 5.5 35 72 7 4.00 12 17 1 5.5 35 42 7 3.81 17 18 5.5 5.5 50 42 7 3.65 16 19 5.5 5.5 20 42 7 3.76 22 20 5.5 5.5 35 42 7 3.61 7 21 3.02905 3.02905 43.2365 58.473 8.0982 2.85 1 22 7.97095 7.97095 26.7635 58.473 5.9018 2.87 18 23 5.5 5.5 35 12 7 3.66 4 24 7.97095 7.97095 43.2365 25.527 5.9018 2.81 21 25 5.5 5.5 35 42 9 1.71 11 26 3.02905 3.02905 26.7635 25.527 5.9018 2.62 The collaborative outcome of different variables can be realized from the perturbation plot which is constructed upon perturbation theory. This theory is based on mathematical methods to find the optimum conditions for solving the issue (Fig. 1b). On the basis of the second-order response surface model, the collaboration of experimental factors including platinum salt solution (ml of 1mmol) (A), ml of inoculum size ( bacillus cereus ) (B), temperature ( ◦ c) (C), time (hours) (D), pH (E) and response (absorbance at 290nm) were exhibited in the form of 3D surface plots (Fig. 2 a-f). The interactive effect of platinum salt solution and inoculum size Figure 1 (a, b) (a) The parity plot showing the association among real and forecast values. (b) Perturbation plot for optimization of biosynthesis of platinum nanoparticles ( bacillus cereus ) with other parameters on the biosynthesis of platinum nanoparticles were studied. An increase in amounts of both causes a sufficient increase in response showing a positive and clear affinity between the two factors however particles get aggregated with an increase in metal concentration [ 27 ]. Similarly, in the presence of high volumes of inoculum size, the response gets weaker as due to large amounts of reducing enzymes, nanoparticles get unstable which may cause their precipitation [ 28 ]. Figure 2 a, c, e, f showed that the biosynthesis of platinum nanoparticles increases with an increase in temperature as response becomes stronger at higher temperature [ 29 ]. An increase in incubation time leads towards an increase in biosynthesis of platinum nanoparticles as Fig. 2 e showed interactive effect of time with temperature. An increase in biosynthesis of platinum nanoparticles was observed from acidic to neutral pH. Maximum biosynthesis of platinum nanoparticles was observed at or close to neutral pH, further, an increase or decrease from neutral point resulted in a decrease in response (Fig. 2 b, d, f) [ 30 ]. It must be understood that the established design has multiple variables and interactive effects of all the variables must be considered. The biosynthesis of platinum nanoparticles was undoubtfully concerned by these variables. It should be noticed that the developed scheme involved multiple variables and the collective effects of all these variables must be given due consideration. The biosynthesis of platinum nanoparticles through bacillus cereus was unquestionably concerned by these variables. For the exceptional recognition of biosynthesis of platinum nanoparticles and to endorse the efficiency and efficacy of the predicted CCD of RSM, experiments were performed in triplicate. Under optimized situations, the experimental and expected values of response (absorbance at 290nm) for the biosynthesis of platinum nanoparticles were in good agreement with each other. The optimized factors for biosynthesis of platinum nanoparticles were platinum salt solution (1mmol) = 3.0ml, inoculum size ( bacillus cereus ) = 3ml, temperature = 43.2 ◦ C, time = 58.4hours, and pH = 8.0 displayed in the form of optimized ramps (Fig. 3 a). The closeness of predicted and the actual results of the proposed model showed its validity. Table 2 ANOVA and statistics for the quadratic model. Source Sum of Squares df Mean Square F-value p-value Model 22.83 20 1.14 26.87 0.0009 significant A-Salt(1mmol) 0.0288 1 0.0288 0.6779 0.4478 B-Inoculum size 0.0450 1 0.0450 1.06 0.3506 C-Temperature 4.03 1 4.03 94.93 0.0002 D-time 0.0392 1 0.0392 0.9228 0.3809 E-ph 0.0008 1 0.0008 0.0188 0.8962 AB 0.3922 1 0.3922 9.23 0.0288 AC 1.75 1 1.75 41.19 0.0014 AD 0.0001 1 0.0001 0.0018 0.9681 AE 0.0315 1 0.0315 0.7421 0.4284 BC 0.0279 1 0.0279 0.6577 0.4542 BD 0.9964 1 0.9964 23.45 0.0047 BE 1.31 1 1.31 30.87 0.0026 CD 0.2251 1 0.2251 5.30 0.0696 CE 0.2142 1 0.2142 5.04 0.0747 DE 1.24 1 1.24 29.18 0.0029 A² 0.0381 1 0.0381 0.8963 0.3873 B² 0.1295 1 0.1295 3.05 0.1413 C² 3.92 1 3.92 92.24 0.0002 D² 0.2471 1 0.2471 5.82 0.0607 E² 12.18 1 12.18 286.62 < 0.0001 Residual 0.2124 5 0.0425 Lack of Fit 0.1194 1 0.1194 5.14 0.0861 not significant Pure Error 0.0930 4 0.0233 Cor Total 23.04 25 Std. Dev. 0.2061 R² 0.9908 Mean 2.96 Adjusted R² 0.9539 C.V. % 6.96 Adequate Precision 15.4515 (a) (b) (c) (d) (e) (f) The color change from yellow to dark brown is visually noticeable (Fig. 3 b) provided an indication for the change of platinum salt into platinum nanoparticles. It was additionally established by the UV-Visible spectrophotometer. UV-1700 pharmaspac UV-Visible spectrophotometer operated at 1nm resolution was used to construct the UV-Visible spectrum. The wavelength range was from 200 to 800nm. A potent surface plasmon response (SPR) response was recognized at 290nm (Fig. 3 c). The absorption response at this wavelength is the distinctive surface plasmon peak of platinum nanoparticles [ 31 ]. At the same time morphology of produced platinum nanoparticles can also be predicted to be isotropic and spherical due to the solitary surface plasmon peak in this region. The produced dark brown mixture was centrifuged at 8000rpm for 15 minutes and dried in air to get the platinum nanoparticles powder. To identify the functional groups responsible for the bio reduction and stabilization of platinum nanoparticles, FTIR (Fourier Transform Infrared) spectra was documented. For this purpose, a Perkin Elmer spectrometer in the range of 4000cm -1 to 500cm -1 was used. Transmission peaks were observed at 3680cm -1 , 3640cm -1 , 3206cm -1 , 2872cm -1 , 2763cm -1 , 1397cm -1 , 1298cm -1 , 1124cm -1 , 979cm -1 , 750cm -1 , and 720cm -1 (Fig. 4 a). Peaks at 3680cm -1 , 3640cm -1 , and 3206cm -1 represent stretching vibrations due to O-H and N-H functional groups. 2872cm -1 is due to C-H stretching of methyl and methylene groups while 2763cm -1 represents N-H and C = O groups of proteins. Peaks from 1400cm -1 to 1000cm -1 are due to C-N symmetrical stretching vibrations of amides, aromatic, and aliphatic amines [ 16 ]. The bands at 750cm -1 , and 720cm -1 signifies the presence of alkyl groups. This data confirms that microbial enzymes are responsible for the bio reduction of platinum ions into platinum nanoparticles. X-ray diffraction pattern of biosynthesized platinum nanoparticles between 2θ values of 25 to 80 is shown in Fig. 4 b. the sharp peaks appeared at 2θ values of 38.6°, 45.35°, 61.15°, and 79.25° with matrix progress orientation along (111), (200), (220), and (311) crystal planes. The results are comparable with standard powder diffraction card of JCPDS, platinum file no. 88-2343. Dynamic Light Scattering Particle Size Analyzer (DLS-PSA) (BT-90 Nanoparticle Size Analyzer) also called photon correlation spectroscopy was used for the analysis of biosynthesized platinum nanoparticles [ 32 ]. The nanoparticle solution is irradiated with electromagnetic beam, which gets scattered due to Brownian motion of the particles and causes their intensity and angle change. Which in turn is adopted to analyze the hydrodynamic diameter of biosynthesized platinum nanoparticles by employing Stokes-Einstein equation. A low concentration aqueous solution of platinum nanoparticles was prepared for analysis. The resulting solution was sonicated for twenty minutes before analysis. The biosynthesized platinum nanoparticles size was found 6.29 to 25.0 nm with a specific surface area of 194.95m 2 /g (Fig. 4 c & d). Scanning electron microscope (SEM) study was carried out on FEI Nova 450 NanoSem. The micrograph elucidates the structure and morphology of the sample. It delivers highly resolved pictures of sample by concentrating on primary, secondary, and backscattered electrons. SEM micrograph of biosynthesized platinum nanoparticles is shown in Fig. 4 e. (a) (b) (c) (b) (c) (d) (e) Antimicrobial activity of biogenic platinum nanoparticles was carried out against four gram positive and four-gram negative bacteria. Gram positive bacterial strains Staphylococcus epidermidis, Staphylococcus aureus, Bacillus subtilis, Bacillus licheniformis and gram-negative bacterial strains Escherichia coli, Salmonella enteria, Porteus mirabilis, Pseudomonas aeruginosa were used. Disc diffusion process was used to ascertain the zones of inhibition (mm) of platinum nanoparticles. Penicillin in concentration of 200µg/ml was used as a standard antibiotic against all the eight bacterial strains. Different zones of inhibitions were noticed for microorganisms based on their structure and composition. However anti-microbial agent was found to be more potent for gram negative bacteria (Fig. 5a). Broth macro dilution method was used to find conventional microbiological parameters like minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) of the bio fabricated PtNPs (Fig. 5b). All experiments were performed in triplicate (Table 3 ). Different reports have been proposed to illustrate the mechanistic aspects of antimicrobial ability of biosynthesized platinum nanoparticles. Due to negative zeta potential of platinum nanoparticles, they may develop chemical interaction with bacterial cell membrane resulting in leakage of cellular contents which is lethal for living cells [ 33 ]. Platinum nanoparticles can penetrate and interact with cellular organelles and biomolecules and thus disturbing the cellular pathways. Similarly antimicrobial agent may cause creation of reactive oxygen species (ROS) within the cell inducing bactericidal oxidative stress [ 34 ]. From this it is established that biosynthesized platinum nanoparticles showed good anti-microbial activity against different microbes and can be used for improvement of new anti-microbials generations, nanomedicines, drug delivery systems, and various more. Table 3 Diameter of zones of inhibition (mm) of biosynthesized PtNPs against standard penicillin, MIC, and MLC (µg/ml) by disc diffusion and macro broth dilution methods respectively. Tested Strains Penicillin (200µg/ml) PtNPs (200µg/ml) MIC (µg/ml) MLC (µg/ml) Staphylococcus epidermidis 20 18 80 140 Staphylococcus aureus 22 10 100 150 Bacillus subtilis 20 15 120 160 Bacillus licheniformis 18 13 150 200 Escherichia coli 25 20 65 100 Salmonella enteria 28 25 100 180 Porteus mirabilis 30 25 120 170 Pseudomonas aeruginosa 20 18 60 120 (a) (b) Figure 5(a, b) (a) Zones of inhibitions of gram-positive and gram-negative bacteria. (b) MIC and MLC of biosynthesized platinum nanoparticles. To examine the antioxidant potential of biosynthesized platinum nanoparticles, DPPH (2,2-diphenyl-1-picrylhydrazyl-hydrate) assay was used. DPPH is purple colored organic compound, undergo a redox reaction, and turned to light yellow in color. On reduction the purple color of DPPH bleached showing antioxidant potential of added sample. The method is simple, sensitive and produces exceptional results for biosynthesized platinum nanoparticles. The antioxidant ability of PtNPs was studied at different time intervals and found that 100µl sample exhibited an excellent radical scavenging activity when compared with Trolox and Butylated hydroxy anisole (BHA) standards (Fig. 6 ). The antioxidant activity of DPPH was represented by the percentage DPPH remaining. It showed significant activity (94.74% remaining) after 30 minutes. Antioxidant potential is mostly related to the size of nanoparticles and size depends on the type of microbe used for the synthesis of nanoparticles. However, it is believed that nanoparticles prepared from gram positive bacteria have better antioxidant activity due to more negative zeta potential [ 35 ]. Microbial methods for synthesis of platinum nanoparticles are superior as microbes have complex and diverse compounds such as enzymes and coenzymes that can only be seen in microbial systems. Biosynthesized platinum nanoparticles were tested for their catalytic potential. The study was performed for disintegration of methyl orange and methylene blue in industrial wastewater. Methyl orange (MO) is an azo dye and is normally used as an indicator. It is orange red in color and shows maximum absorbance at 460nm. Methylene blue [ 6 ] is a thiazine dye that finds extensive uses. It is deep blue in color and has maximum absorbance at 665nm. Even if sodium borohydride is a strong reducing agent but still is unable to accomplish the reduction of both dyes due to great difference redox potentials of dyes and reducing agent. These reactions are thermally favorable but kinetically unfavorable. The degradation study of methyl orange, and methylene blue with NaBH 4 was carried out in presence and absence of biosynthesized platinum nanoparticles. The blank experiments were performed without platinum nanoparticles exhibited no color change even after 30minutes so there was no degradation of both dyes in the absence of platinum nanoparticles. At the same time, degradation occurred rapidly by adding small amounts of platinum nanoparticles which showed their catalytic potential. The degradation is evident by fading the orange red and deep blue colors of methyl orange and methylene blue as well as decrease in their absorbance at 460nm and 665nm respectively. Spectrophotometer was used to study reaction kinetics (Table 4 ). The NaBH 4 added is much higher in amount as compared to the dyes, practically its concentration remains constant throughout the course of reaction. So, the kinetics followed by both the reactions were pseudo first order. The concentration of both dyes altered with time. This become clear in (Fig. 7 a, b) which are plots of ln[A] with time for degradation of methyl orange and methylene blue. The process is a redox reaction. The reaction between NaBH 4 and dyes is an electron transfer mechanism in which NaBH 4 is a donor and dye act as accepter. The added platinum nanoparticles have potential between these two and make the transmission of electron easy. Due to the large surface area of platinum nanoparticles, the rate of reduction process increased. Before the onset of reduction, both the dye and borohydride ions (BH 4 − ) get adsorbed on the surface of catalyst. The borohydride ions release electrons to the surface of catalyst from which dye molecules gain electrons and get reduced. Due to this reason, presence of nano catalyst enhances the rate of reduction process and proved to be an efficient catalyst for the removal of harmful dyes from industrial wastewater [ 36 ]. The illegal dumping of waste from industries has rebellious pollutants, which is a worldwide problem with more severity in developing nations. Table 4 Deterioration of methyl orange and methylene blue in presence and absence of biosynthesized platinum nanoparticles along with ln [A] calculated from pseudo first order kinetics. Dye Time (min) Absorbance with PtNPs ln[A] Absorbance without PtNPs ln[A] Methyl orange 0 5 10 15 20 25 30 2.64 2.58 2.10 1.72 1.29 0.87 0.31 0.97 0.94 0.74 0.54 0.25 -0.14 -1.17 2.64 2.62 2.55 2.51 2.48 2.44 2.40 0.97 0.96 0.93 0.92 0.91 0.89 0.87 Methylene blue 0 5 10 15 20 25 30 2.90 2.81 1.85 1.39 0.81 0.55 0.06 1.06 1.03 0.61 0.33 -0.21 -0.59 -2.81 2.90 2.87 2.54 2.01 1.86 1.74 1.57 1.06 1.05 0.93 0.67 0.62 0.55 0.45 (a) (b) Materials and Methods Materials All the chemicals including hexa-chloroplatinic acid H 2 PtCl 6 (99.9%), nutrient broth, agar, DPPH (2,2-diphenyl-1-picrylhydrazyl-hydrate), methyl orange, and methylene blue used in this examination were of analytical grade and procured from Sigma Aldrich Chemicals Germany. Double distilled deionized water was used during this examination. The bacillus cereus was isolated from dirty and filthy soil and then decontaminated and identified microbiologically. The pure cultures were held in reserve and re-cultured from time to time in nutrient broth agar medium slants. Staphylococcus epidermidis, Staphylococcus aureus, Bacillus subtilis, Bacillus licheniformis, Escherichia coli, Salmonella enteria, Porteus mirabilis , and Pseudomonas aeruginosa were attained from Mayo Hospital Lahore for antimicrobial activity. Inoculum Size Preparation Conical flask (250ml) containing 1.3g nutrient broth per 100ml of buffer solution (pH 5 to 9) was autoclaved at 121 ° C for 30minutes. A loop full of Bacillus cereus was then injected into the autoclaved flask and placed in a shaking incubator at 37 ◦ C, 120rpm for 24 hours. Green synthesis of Platinum Nanoparticles Production of platinum nanoparticles was led by performing all the twenty-six experiments designed through CCD model of RSM (Table 1 ). The experiments were carried out by mixing 1 to 10ml of platinum salt solution with 1 to 10ml inoculum of bacillus cereus ranging in pH from 5 to 9 and placed in incubator shaker for specified time of 12 to 72 hours at temperature of 25 ° C to 50 ° C and 120rpm. The response of each experiment was monitored through UV-visible spectrophotometer by measuring absorbance at 290nm. Each experiment was performed in triplicate. After that, optimized factors for maximum biosynthesis of platinum nanoparticles were obtained (Fig. 3 a). One to one ratio of platinum salt solution (1millimolar) and inoculum size ( bacillus cereus ) maintained at pH eight were mixed and placed in incubator shaker for 58.4hours at 43.2 ° C and 120rpm. The color change from yellow to dark brown was observed (Fig. 3 b). The biosynthesized platinum nanoparticles were separated from reaction mixture through centrifugation for ten minutes at 8000rpm and washed with deionized water for the elimination of active molecules. The nanoparticles were air dried for further studies. Antimicrobial activity Gram positive bacterial strains Staphylococcus epidermidis, Staphylococcus aureus, Bacillus subtilis, Bacillus licheniformis and gram-negative bacterial strains Escherichia coli, Salmonella enteria, Porteus mirabilis, Pseudomonas aeruginosa were used to find the zone of inhibition, minimum inhibitory concentrations (MIC), and minimum lethal concentrations (MLC) of biosynthesized platinum nanoparticles. Kirby-Bauer disc diffusion method [ 37 ] was employed to calculate the zone of inhibitions of the biosynthesized platinum nanoparticles. Each of the bacterial culture was swabbed in the sterilized petri plate with Mueller Hinton agar. Into each petri plate, two discs (6mm), one soaked with standard antibiotic Penicillin (200µg/ml) and second saturated with biosynthesized platinum nanoparticles (200µg/ml), were placed on the uppermost layer. These petri plates were placed in an incubator for 24hours at the temperature of 37 ° C. After that diameter (mm) of zone of inhibition (Table 3 ) was measured for each bacterial species (Fig. 5a). The minimum inhibitory concentration (MIC) represents the lowest concentration of platinum nanoparticles that inhibits the growth (99%) of microbes. Broth macro dilution method was used to find the MIC. To encounter MIC, the Lauria Bertain (LB) broth and two times chronological dilutions of biosynthesized platinum nanoparticles (1600µg/ml to 0.05µg/ml) were used. A test tube with microbe in LB broth acts as positive control while a negative control tube contains LB broth only. The positive and negative controls were also carried out in parallel. All these were placed in an orbital shaker at 37 ° C and 120rpm for 24 hours and after that MIC was determined. For the determination of minimum lethal concentration (MLC), 25µl of each clear tube with no visible bacterial growth was seeded on Muller-Hinton Agar (MHA) plates and placed in an incubator at 37 ° C for 24hours. The MLC represents the lowest concentration of biosynthesized platinum nanoparticles which assassinate 100% of microbial growth so there will be no growth on MHA plates (Table 3 ). Bactericidal effects were represented by MLC while MIC represents the bacteriostatic effects of biosynthesized platinum nanoparticles against the tested microbial strains (Fig. 5b). Antioxidant Potential For DPPH assay, a standard solution of DPPH in methanol was prepared by dissolving 25mg/l. the absorbance of UV-Visible spectrophotometer was adjusted at 1.00 ± 0.02 at 515nm. The solution of bio fabricated platinum nanoparticles was processed in double distilled water by adding 3mg/ml. For antioxidation potential, to the 2.5 ml of methanolic DPPH solution was added 100µl of biosynthesized platinum nanoparticles solution and mixed vigorously. The mixture could stand for 30 minutes in dark. Measure the absorbance after every 5 minutes up to 30 minutes. The scavenging ability was calculated by following expression, % DPPH remaining = (DPPH)t = t/ (DPPH)t = 0 × 100 where (DPPH)t = t was absorbance of DPPH solution at a specific time and (DPPH)t = 0 showed absorbance of DPPH solution at the initial time. Catalytic Degradation of dyes Catalytic disintegration of methyl orange (MO) and methylene blue [ 6 ] through NaBH 4 was investigated when biosynthesized platinum nanoparticles act as catalyst. The reaction was performed by mixing 2ml aqueous solution (0.01×10 − 2 ) of each of the dyes with 0.5ml (0.006M) solution of sodium borohydride followed by 0.5ml (0.05mg/ml) of biosynthesized platinum nanoparticles. The progress of two reactions and change in concentration of methyl orange and methylene blue with time was measured via UV-visible spectrophotometer at 460nm and 665nm respectively. Figure out the change in absorbance of each solution after every 5 minutes delay for 30 minutes. Absorbance is directly proportional to concentration, so a decrease in concentration results a decrease in absorption (Table 4 ). Two blank reactions for each dye without biosynthesized platinum nanoparticles were also performed. For representation of reduction of methyl orange and methylene blue in presence and absence of platinum nanoparticles plots of ln[A] along time are shown in (Fig. 7 a, b). CONCLUSION Resultantly, bacillus cereus was utilized for the biogenic synthesis of platinum nanoparticles. Central composite design (CCD) of response surface methodology (RSM) was used for the optimization of reaction conditions such as metal salt concentration, inoculum size, temperature, time, and pH. On the basis of ANOVA results for quadratic model, three dimensional responses, and perturbation plot optimized conditions for small sized, high surface area, biosynthesized platinum nanoparticles are, platinum salt solution (1mmol) = 3.0ml, inoculum size ( bacillus cereus ) = 3ml, temperature = 43.2 ◦ C, time = 58.4hours, and pH = 8.0. By following these set of optimized conditions, the biosynthesized platinum nanoparticles were of size 6.29 to 25.0nm with 194.95m 2 /g surface area. The CCD of RSM detected as a vital tool to explain the distinct and combined properties of various tentative data points and their optimization for improved biosynthesis of platinum nanoparticles in a cost effective and time effective method. Uv-visible spectrophotometer, FTIR spectroscopy, DLS-particle size analyzer, XRD, and SEM were used for characterization of biosynthesized platinum nanoparticles. For antibacterial activity, four gram-positive bacterial strains Staphylococcus epidermidis, Staphylococcus aureus, Bacillus subtilis, Bacillus licheniformis and four gram-negative bacterial strains Escherichia coli, Salmonella enteria, Porteus mirabilis, Pseudomonas aeruginosa were used with standard antibiotic Penicillin to calculate the zones of inhibitions by disc diffusion method and MIC and MLC by broth macro dilution method. The biosynthesized platinum nanoparticles were also evaluated for their antioxidant potential through DPPH (2,2-diphenyl-1-picrylhydrazyl) assay. Catalytic role of biosynthesized platinum nanoparticles was assessed for disintegration of methyl orange and methylene blue with time and give brilliant results. It is first ever report in which platinum nanoparticles were synthesized through bacillus cereus and optimization of experimental conditions were carried out through CCD of RSM. The biosynthesized platinum nanoparticles were also studied for their biomedical and environmental applications. Disclosure Declaration The authors have no conflict of interests that can impact the reported research work Abbreviations CCD Central Composite Design RSM Response Surface Methodology NPs nanoparticles PtNPs platinum nanoparticles FTIR Fourier transform infrared spectroscopy XRD x-ray diffraction DLS Dynamic light scattering SEM Scanning Electron Microscopy MIC Minimum Inhibitory Concentration MLC Minimum Lethal Concentration BHA Butylated hydroxy anisole DPPH 2,2-diphenyl-1-picrylhydrazyl MO Methyl orange MB Methylene blue Declarations Ethical Approval This item does not comprise any studies with humans or animals made by any of the authors. Funding Declaration This research received no external funding or financial support. Author Contribution S I performed experiments and write the manuscript. Z A provide assistance in experiments. S I and M M prepare all the figures. 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Dauthal, P., & Mukhopadhyay, M. (2015). Biofabrication, characterization, and possible bio-reduction mechanism of platinum nanoparticles mediated by agro-industrial waste and their catalytic activity. Journal of Industrial and Engineering Chemistry , 22 , 185–191. Berne, B. J., & Pecora, R. (2000). Dynamic light scattering: with applications to chemistry, biology, and physics . Courier Corporation. Slavin, Y. N., et al. (2017). Metal nanoparticles: understanding the mechanisms behind antibacterial activity. Journal of nanobiotechnology , 15 , 1–20. Eramabadi, P., et al. (2020). Microbial cell lysate supernatant (CLS) alteration impact on platinum nanoparticles fabrication, characterization, antioxidant and antibacterial activity. Materials Science and Engineering: C , 117 , 111292. Daré, R. G., & Lautenschlager, S. O. (2025). Nanoparticles with Antioxidant Activity . MDPI. p. 221. Ibrahim, S., et al. (2021). Optimization for biogenic microbial synthesis of silver nanoparticles through response surface methodology, characterization, their antimicrobial, antioxidant, and catalytic potential. Scientific Reports , 11 (1), 770. Drew, W. L., et al. (1972). Reliability of the Kirby-Bauer disc diffusion method for detecting methicillin-resistant strains of Staphylococcus aureus. Applied microbiology , 24 (2), 240–247. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6922293","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":483165009,"identity":"40c572fd-d66e-4a0d-b09c-93185304d23c","order_by":0,"name":"Saba Ibrahim","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA4UlEQVRIiWNgGAWjYFACHgjFD6GYgZiNSC2SDSRrMThArBb59rMHPzD8skvcfH7xMwmGCuvEBum2BLxaDM7kJUsw9iUnbrvxzEyC4Ux6YoPMsQP4tTDkGEgw9hwAajnDJsHYdjixQSK9Ab/D+t8Y/wBp2TwDpOUfEVoYbuQA3fPjQOIG/h6glgaQljQCDrvxLs2CsSHZeMYNNmOLhGPpxm0yxxIIOCz38A2GP3ay/f2HH974UGMt2y/dZoDfYUDA/LcNSEoAzQYZzyZBUAMI/AFifpgPiNMyCkbBKBgFIwgAAGHtSTAq29kaAAAAAElFTkSuQmCC","orcid":"","institution":"Government College University","correspondingAuthor":true,"prefix":"","firstName":"Saba","middleName":"","lastName":"Ibrahim","suffix":""},{"id":483165010,"identity":"16902cd9-79c7-4f4e-9c85-0ae926d1efeb","order_by":1,"name":"Zahoor Ahmad","email":"","orcid":"","institution":"University of Engineering and Technology","correspondingAuthor":false,"prefix":"","firstName":"Zahoor","middleName":"","lastName":"Ahmad","suffix":""},{"id":483165013,"identity":"68328b8d-b3ca-4c50-b03a-8747fadb2d01","order_by":2,"name":"Muhammad Mujahid","email":"","orcid":"","institution":"Universiti Sains Malaysia","correspondingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"","lastName":"Mujahid","suffix":""}],"badges":[],"createdAt":"2025-06-18 10:53:30","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6922293/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6922293/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":86447781,"identity":"1738c68b-152e-46df-b057-d5b2f20412c6","added_by":"auto","created_at":"2025-07-10 18:09:15","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":41859,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(a, b):\u003c/strong\u003e (a) The parity plot showing the association among real and forecast values. (b) Perturbation plot for optimization of biosynthesis of platinum nanoparticles\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6922293/v1/f94ffc96701f4c62c1634b0a.jpg"},{"id":86447783,"identity":"2b755495-663a-471f-bae0-fd3fa64e957e","added_by":"auto","created_at":"2025-07-10 18:09:15","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":212502,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(a-f):\u003c/strong\u003e Three-dimensional response surface plots indicating mutual interaction and individual effects of experimental variables on biogenesis of PtNPs (a) Platinum solution (ml) and Temperature (\u003csup\u003e◦\u003c/sup\u003eC) (b) Platinum solution (ml) and pH (c) Inoculum size (ml) and Temperature \u003csup\u003e(◦\u003c/sup\u003eC) (d) Inoculum size (ml) and pH, (e) Temperature (\u003csup\u003e◦\u003c/sup\u003eC) and Time (hours), (f) Temperature (\u003csup\u003e◦\u003c/sup\u003eC) and pH.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6922293/v1/672d097ae4b0f07e0a245eb9.png"},{"id":86448091,"identity":"f9202ca2-09b6-48a8-ab9d-39eb4b98e38f","added_by":"auto","created_at":"2025-07-10 18:17:15","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":219563,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(a, b, c):\u003c/strong\u003e (a) Optimized ramps showing optimized results of platinum salt solution, inoculum size, temperature, time, and pH for the biosynthesis of PtNPs. (b) Visual color change for biological synthesis of platinum nanoparticles, A = before reaction and B = after reaction. (c) A UV-visible spectrum of biosynthesized PtNPs through \u003cem\u003ebacillus cereus.\u003c/em\u003e\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6922293/v1/afebf07bdd0dec137445e03e.png"},{"id":86447791,"identity":"724ab609-5581-4855-a139-804ec8b8ac6d","added_by":"auto","created_at":"2025-07-10 18:09:15","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":194666,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(a, b, c, d, e): (a) \u003c/strong\u003eFTIR spectra \u003cstrong\u003e(b)\u003c/strong\u003e X-ray diffraction pattern \u003cstrong\u003e(c)\u003c/strong\u003eDLS pattern \u003cstrong\u003e(d)\u003c/strong\u003e size distribution (e) SEM images of biosynthesized platinum nanoparticles.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6922293/v1/659ee298b6bebd65fbaf1576.png"},{"id":86448090,"identity":"30b7e8bc-8041-4736-bb1d-36975de25a30","added_by":"auto","created_at":"2025-07-10 18:17:15","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":41645,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(a, b):\u003c/strong\u003e (a) Zones of inhibitions of gram-positive and gram-negative bacteria. (b) MIC and MLC of biosynthesized platinum nanoparticles.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-6922293/v1/847723547a32157cd5d6c723.png"},{"id":86447785,"identity":"75698893-4a0e-4493-a73b-7efb4c50aa1f","added_by":"auto","created_at":"2025-07-10 18:09:15","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":128584,"visible":true,"origin":"","legend":"\u003cp\u003eDPPH assay of biosynthesized PtNPs.\u003c/p\u003e","description":"","filename":"6.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6922293/v1/9c7dc8597cb5048bf5674014.jpg"},{"id":86448635,"identity":"a9fa644a-142e-4e3f-9893-72577a7064c3","added_by":"auto","created_at":"2025-07-10 18:25:15","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":59499,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(a, b):\u003c/strong\u003e Dye degradation studies (a) methyl orange, (b) methylene blue in presence and absence of biosynthesized platinum nanoparticles.\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-6922293/v1/366d9a7bfd30609c67752b66.png"},{"id":88510481,"identity":"ed425562-b82a-4796-9010-c473ecda2029","added_by":"auto","created_at":"2025-08-07 07:55:05","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1959188,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6922293/v1/aae0bbfe-1a2e-4b56-aa55-1f602d36edec.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Central Composite Design for Optimizing the Biogenic fabrication of Platinum Nanoparticles Using bacillus cereus and TheirAntimicrobial, Antioxidant, and catalytic Potential","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eMicrobial synthesis of metal nanoparticles is of colossal concern to modern nanotechnology as it is simple, cost-effective, and eco-friendly. In the present study, a novel green method for biogenic synthesis of platinum nanoparticles has been developed using a gram-positive bacterial strain [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Green synthesis of metal nanoparticles is attaining extraordinary consideration in various applications [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e] [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. To synthesize metal nanosized particles, various methods have been used, including colloidal systems and chemical reduction [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. These approaches are favored in research due to their simplicity, reliability, and strong potential for biomedical applications, while maintaining the structural integrity of the nanoparticles [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Furthermore, the preparation of nanoparticles from transition metals has a diversity of functions including material science, chemistry, and medicine[\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. Transition metal nanoparticles are vital due to their biocompatibility and eco-friendly versatile nature. Nanoparticles of Pb, Cu, Pd, Ag, Au, and Pt find applications in catalysis, pollution remediation, drug delivery, wastewater [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. The choice of method for synthesis of metal nanoparticles is very important as during synthesis proceedings like kinetic interaction of the metal with reducing agent, retention of the stabilizing agent with metal nanoparticles and distinct experimental approaches causes a robust effect on constancy, morphology, physical and chemical properties [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Among these metal nanoparticles, the synthesis of platinum nanoparticles, in particular, their biogenic fabrication has attracted great interest as it is sustainable, green, and cost-effective. Platinum nanoparticles have remarkable applications due to their large surface area, corrosion, and chemical resistance. Platinum nanoparticles are used as dental material ingredient [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e], coating of medical devices [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e], water treatment [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e], cancer treatment [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e], antiaging [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], cosmetics [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e], dye degradation [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e], and much more. Generally, two systems are used for the biogenic fabrication of metal nanoparticles. First is the use of different parts of plants like roots, stems, leaf, fruit and flower [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e], [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Plants have rich amounts of secondary metabolites which are responsible for reduction of metal ion to metal nanoparticles. However, concentration of these secondary metabolites is different in different parts of plants and methods of extraction influence their concentration [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e] so the quality and quantity of nanoparticles get compromised. Second is microbial system which includes bacteria, fungi, algae, actinomycetes, yeast and are more rapid. Microorganisms have the ability to produce metal nanoparticles intracellularly and extracellularly. In both, reductase enzyme is responsible for conversion of metal ion into nanoparticles [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. However, microbial systems have been less focused for synthesis of platinum nanoparticles.\u003c/p\u003e\u003cp\u003eIn the present work, a novel microbial synthesis of platinum nanoparticles through \u003cem\u003ebacillus cereus\u003c/em\u003e has been reported. As conditions of an experiment such as pH, temperature, the concentration of metal salt, time of reaction, inoculum size of \u003cem\u003ebacillus cereus\u003c/em\u003e, play a vital role in the biosynthesis of platinum nanoparticles. A manual set up to study one aspect at a time by making others persistent for these experimental conditions is long, tedious, time-consuming, costly, and causes errors so it is avoided. Response surface methodology (RSM) is a blend of mathematical and statistical procedures used for designing experiments, construction of models, assessing the operative factors, and most significant is searching for the optimal conditions for a required response [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. This has to be done by considering three main stepladders as a performance of statistically planned experiments, approximating the coefficient in the mathematical model, and forecasting the response by checking the suitability of the model [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. We used central composite design (CCD) of response surface methodology (RSM) which is mostly adapted for optimization of multiple factors varying simultaneously. It can provide comprehensive intuition of direct, curved, and pair-wise mutual effects of variables on response. The central composite design is a more efficient, available operative tool for the optimization of complex processes [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Few reports are available regarding the synthesis of platinum nanoparticles through microbes[\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e], [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. However, no report is available showing RSM mediated optimization of biosynthesis of platinum nanoparticles through \u003cem\u003ebacillus cereus\u003c/em\u003e. In the current study, \u003cem\u003ebacillus cereus\u003c/em\u003e has been used to mediate the biosynthesis of platinum nanoparticles through the reduction of ionic platinum into metallic nano platinum. The procedure of biosynthesis was optimized via the central composite design technique in ordered to get the proper biosynthetic conditions and understanding their interactive effects. Platinum nanoparticles were characterized through simple systematic techniques of UV-visible spectrophotometer, scanning electron microscopy (SEM), Furrier transform infra-red (FTIR) spectroscopy, Dynamic light scattering-particle size analyzer (DLS-PSA), and X-ray diffraction (XRD). The present study is planned to regulate methods for bacterial synthesis of small-sized, stable, and mono-dispersed platinum nanoparticles having biomedical and environmental applications.\u003c/p\u003e"},{"header":"RESULTS AND DISCUSSION","content":"\u003cp\u003eIn the present work, central composite design (CCD) of response surface methodology (RSM) was employed to find the optimized experimental conditions of (A) metal salt concentration (ml of 1mM), (B) inoculum size in ml (\u003cem\u003ebacillus cereus\u003c/em\u003e), (C) temperature, (D) time, and (E) pH for green synthesis of platinum nanoparticles. Response Surface Methodology Design-Expert-12 software was employed for the purpose that can be accessed by the link \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.statease.com/software/design-expert/\u003c/span\u003e\u003cspan address=\"https://www.statease.com/software/design-expert/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. The central composite design gave a model of twenty-six experiments and the response of each experiment was represented as absorbance at 290nm (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The design was used to attain the best surface plasmon response of synthesized platinum nanoparticles with reciprocal peak intensity. Basically, the process of optimization through CCD of RSM can be achieved by performing statistically designed experiments, then assessing the design coefficients, and finally examining the design fitness [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. After performing all the twenty-six experiments, results in the form of coded factors were presented by a polynomial quadratic equation.\u003c/p\u003e\u003cp\u003eSPR Intensity\u0026thinsp;=\u0026thinsp;+\u0026thinsp;3.94\u0026thinsp;+\u0026thinsp;0.0659 A\u0026thinsp;+\u0026thinsp;0.0824 B\u0026thinsp;+\u0026thinsp;0.7797 C \u0026minus;\u0026thinsp;0.0769 D -0.0110 E\u0026thinsp;+\u0026thinsp;0.3582 AB \u0026ndash; 0.7566 AC\u0026thinsp;+\u0026thinsp;0.0050 AD\u0026thinsp;+\u0026thinsp;0.1016 AE \u0026ndash; 0.0956 BC\u0026thinsp;+\u0026thinsp;0.5710 BD\u0026thinsp;+\u0026thinsp;0.6551 BE \u0026ndash; 0.2714 CD \u0026ndash; 0.2648 CE\u0026thinsp;+\u0026thinsp;0.6368 DE \u0026ndash; 0.0429 A\u003csup\u003e2\u003c/sup\u003e \u0026ndash; 0.0790 B\u003csup\u003e2\u003c/sup\u003e \u0026ndash; 0.4348 C\u003csup\u003e2\u003c/sup\u003e \u0026ndash; 0.1092 D\u003csup\u003e2\u003c/sup\u003e \u0026ndash; 0.7665 E\u003csup\u003e2\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eThis coded factor equation helps to make estimates about the retort for given stages of each factor. The symbiotic and combative effects of these factors on the feedback were represented with positive and negative symbols. The higher levels of the factors are coded as +\u0026thinsp;1 and lower levels are coded as -1. The coded equation is beneficial in the identification of the relative effect of the factors by equating the factor coefficients. Based on this equation, the parity plot explains the proper correlation between actual and forecast responses. The difference between actual and predicted response is proportional to the space of these values from the slanted line (Fig.\u0026nbsp;1a). The level of significance of each factor in the regression model in terms of linear, quadratic, and interactive factors, was determined by p-values. The integrity of the present model was checked by the determinant coefficient (R\u003csup\u003e2\u003c/sup\u003e) which justifies the fluctuation in the predicted and actual response. The R\u003csup\u003e2\u003c/sup\u003e of 0.9908 exhibits that 99.08% variation in platinum nanoparticle synthesis is because of independent variables. The model R\u003csup\u003e2\u003c/sup\u003e is in close agreement with adjusted R\u003csup\u003e2\u003c/sup\u003e which is 0.9539. For a good model, R\u003csup\u003e2\u003c/sup\u003e should be in the range of 0\u0026ndash;1[\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Adequate precision means a signal to noise ratio. A value bigger than 4 is needed. The value of adequate precision 15.4515 stated satisfactory signals. The f-value of 5.14 and p-value of 0.0861 of non-significant lack of fit recommend that the adopted regression model is significant. The outcomes of the analysis of variance (ANOVA) are fundamental to evaluate the implication, accuracy, and authenticity of the statistical model and are given in (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The model f-value 26.87 and model p-value 0.0009 suggest that the model and model terms C, AB, AC, BD, BE, DE, C\u003csup\u003e2\u003c/sup\u003e, E\u003csup\u003e2\u003c/sup\u003e are highly recognizable. A comparatively lower value of CV (variation coefficient) 6.96% exhibits the efficiency, credibility, and accuracy of model trials.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eThe central composite design (CCD) of the experiments for the biosynthesis of platinum nanoparticles through \u003cem\u003ebacillus cereus\u003c/em\u003e.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"9\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" 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colname=\"c6\"\u003e\u003cp\u003eFactor 4\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eFactor 5\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eResponse\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eStd\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRun\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003eA: Salt (1mmol)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003eB: Inoculum Size\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003eC: Temperature\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eD: time\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eE: pH\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eAbsorbance\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003emL\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003emL\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e\u003csup\u003e◦\u003c/sup\u003eC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003eHours\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003eMol/L of H\u003csup\u003e+\u003c/sup\u003e ions\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" 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colname=\"c3\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3.96\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3.80\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e7.97095\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.02905\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e43.2365\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e25.527\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e8.0982\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2.33\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e8\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e3.02905\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e7.97095\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e26.7635\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e58.473\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e8.0982\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2.37\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e7.97095\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e7.97095\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e26.7635\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e25.527\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e8.0982\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2.57\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e26\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e14\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3.71\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1.56\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e16\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e72\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e4.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3.81\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e50\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3.65\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e16\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3.76\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e22\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3.61\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e21\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e3.02905\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.02905\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e43.2365\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e58.473\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e8.0982\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2.85\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e22\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e7.97095\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e7.97095\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e26.7635\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e58.473\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e5.9018\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2.87\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e23\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e7\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e3.66\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e7.97095\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e7.97095\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e43.2365\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e25.527\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e5.9018\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2.81\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e21\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e5.5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e35\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e42\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e9\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e1.71\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e26\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e3.02905\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.02905\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e26.7635\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e25.527\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e5.9018\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e2.62\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eThe collaborative outcome of different variables can be realized from the perturbation plot which is constructed upon perturbation theory. This theory is based on mathematical methods to find the optimum conditions for solving the issue (Fig.\u0026nbsp;1b). On the basis of the second-order response surface model, the collaboration of experimental factors including platinum salt solution (ml of 1mmol) (A), ml of inoculum size (\u003cem\u003ebacillus cereus\u003c/em\u003e) (B), temperature (\u003csup\u003e◦\u003c/sup\u003ec) (C), time (hours) (D), pH (E) and response (absorbance at 290nm) were exhibited in the form of 3D surface plots (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ea-f). The interactive effect of platinum salt solution and inoculum size\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eFigure 1 (a, b)\u003c/strong\u003e\u003cp\u003e(a) The parity plot showing the association among real and forecast values. (b) Perturbation plot for optimization of biosynthesis of platinum nanoparticles\u003c/p\u003e\u003c/p\u003e\u003cp\u003e(\u003cem\u003ebacillus cereus\u003c/em\u003e) with other parameters on the biosynthesis of platinum nanoparticles were studied. An increase in amounts of both causes a sufficient increase in response showing a positive and clear affinity between the two factors however particles get aggregated with an increase in metal concentration [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. Similarly, in the presence of high volumes of inoculum size, the response gets weaker as due to large amounts of reducing enzymes, nanoparticles get unstable which may cause their precipitation [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Figure\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ea, c, e, f showed that the biosynthesis of platinum nanoparticles increases with an increase in temperature as response becomes stronger at higher temperature [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. An increase in incubation time leads towards an increase in biosynthesis of platinum nanoparticles as Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003ee showed interactive effect of time with temperature. An increase in biosynthesis of platinum nanoparticles was observed from acidic to neutral pH. Maximum biosynthesis of platinum nanoparticles was observed at or close to neutral pH, further, an increase or decrease from neutral point resulted in a decrease in response (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e2\u003c/span\u003eb, d, f) [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. It must be understood that the established design has multiple variables and interactive effects of all the variables must be considered. The biosynthesis of platinum nanoparticles was undoubtfully concerned by these variables.\u003c/p\u003e\u003cp\u003eIt should be noticed that the developed scheme involved multiple variables and the collective effects of all these variables must be given due consideration. The biosynthesis of platinum nanoparticles through \u003cem\u003ebacillus cereus\u003c/em\u003e was unquestionably concerned by these variables. For the exceptional recognition of biosynthesis of platinum nanoparticles and to endorse the efficiency and efficacy of the predicted CCD of RSM, experiments were performed in triplicate. Under optimized situations, the experimental and expected values of response (absorbance at 290nm) for the biosynthesis of platinum nanoparticles were in good agreement with each other. The optimized factors for biosynthesis of platinum nanoparticles were platinum salt solution (1mmol)\u0026thinsp;=\u0026thinsp;3.0ml, inoculum size (\u003cem\u003ebacillus cereus\u003c/em\u003e)\u0026thinsp;=\u0026thinsp;3ml, temperature\u0026thinsp;=\u0026thinsp;43.2\u003csup\u003e◦\u003c/sup\u003eC, time\u0026thinsp;=\u0026thinsp;58.4hours, and pH\u0026thinsp;=\u0026thinsp;8.0 displayed in the form of optimized ramps (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003ea). The closeness of predicted and the actual results of the proposed model showed its validity.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eANOVA and statistics for the quadratic model.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"7\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSource\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eSum of Squares\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003edf\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eMean Square\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eF-value\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003ep-value\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eModel\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e22.83\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.14\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e26.87\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.0009\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003esignificant\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eA-Salt(1mmol)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.0288\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.0288\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.6779\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.4478\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eB-Inoculum size\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.0450\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.0450\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e1.06\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.3506\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eC-Temperature\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4.03\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e4.03\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e94.93\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.0002\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eD-time\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.0392\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.0392\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.9228\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.3809\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eE-ph\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.0008\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.0008\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.0188\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.8962\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAB\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.3922\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.3922\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e9.23\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.0288\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.75\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.75\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e41.19\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.0014\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAD\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.0018\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.9681\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAE\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.0315\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.0315\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.7421\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.4284\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBC\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.0279\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.0279\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.6577\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.4542\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBD\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.9964\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.9964\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e23.45\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.0047\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBE\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.31\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.31\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e30.87\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.0026\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCD\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.2251\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.2251\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5.30\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.0696\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCE\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.2142\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.2142\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5.04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.0747\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDE\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e29.18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.0029\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eA\u0026sup2;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.0381\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.0381\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e0.8963\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.3873\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eB\u0026sup2;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.1295\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.1295\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e3.05\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.1413\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eC\u0026sup2;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e3.92\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e3.92\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e92.24\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.0002\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eD\u0026sup2;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.2471\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.2471\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5.82\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.0607\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eE\u0026sup2;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e12.18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e12.18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e286.62\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eResidual\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.2124\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.0425\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLack of Fit\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.1194\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.1194\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e5.14\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.0861\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003enot significant\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePure Error\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.0930\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.0233\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCor Total\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e23.04\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eStd. Dev.\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0.2061\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"3\" nameend=\"c5\" namest=\"c3\"\u003e\u003cp\u003e\u003cb\u003eR\u0026sup2;\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.9908\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"1\" nameend=\"c7\" namest=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eMean\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2.96\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"3\" nameend=\"c5\" namest=\"c3\"\u003e\u003cp\u003e\u003cb\u003eAdjusted R\u0026sup2;\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.9539\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"1\" nameend=\"c7\" namest=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eC.V. %\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.96\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"3\" nameend=\"c5\" namest=\"c3\"\u003e\u003cp\u003e\u003cb\u003eAdequate Precision\u003c/b\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e15.4515\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colspan=\"1\" nameend=\"c7\" namest=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e(a) (b)\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003e(c) (d)\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e(e) (f)\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eThe color change from yellow to dark brown is visually noticeable (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003eb) provided an indication for the change of platinum salt into platinum nanoparticles. It was additionally established by the UV-Visible spectrophotometer. UV-1700 pharmaspac UV-Visible spectrophotometer operated at 1nm resolution was used to construct the UV-Visible spectrum. The wavelength range was from 200 to 800nm. A potent surface plasmon response (SPR) response was recognized at 290nm (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003ec). The absorption response at this wavelength is the distinctive surface plasmon peak of platinum nanoparticles [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. At the same time morphology of produced platinum nanoparticles can also be predicted to be isotropic and spherical due to the solitary surface plasmon peak in this region. The produced dark brown mixture was centrifuged at 8000rpm for 15 minutes and dried in air to get the platinum nanoparticles powder. To identify the functional groups responsible for the bio reduction and stabilization of platinum nanoparticles, FTIR (Fourier Transform Infrared) spectra was documented. For this purpose, a Perkin Elmer spectrometer in the range of 4000cm\u003csup\u003e-1\u003c/sup\u003e to 500cm\u003csup\u003e-1\u003c/sup\u003e was used. Transmission peaks were observed at 3680cm\u003csup\u003e-1\u003c/sup\u003e, 3640cm\u003csup\u003e-1\u003c/sup\u003e, 3206cm\u003csup\u003e-1\u003c/sup\u003e, 2872cm\u003csup\u003e-1\u003c/sup\u003e, 2763cm\u003csup\u003e-1\u003c/sup\u003e, 1397cm\u003csup\u003e-1\u003c/sup\u003e, 1298cm\u003csup\u003e-1\u003c/sup\u003e, 1124cm\u003csup\u003e-1\u003c/sup\u003e, 979cm\u003csup\u003e-1\u003c/sup\u003e, 750cm\u003csup\u003e-1\u003c/sup\u003e, and 720cm\u003csup\u003e-1\u003c/sup\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e4\u003c/span\u003ea). Peaks at 3680cm\u003csup\u003e-1\u003c/sup\u003e, 3640cm\u003csup\u003e-1\u003c/sup\u003e, and 3206cm\u003csup\u003e-1\u003c/sup\u003e represent stretching vibrations due to O-H and N-H functional groups. 2872cm\u003csup\u003e-1\u003c/sup\u003e is due to C-H stretching of methyl and methylene groups while 2763cm\u003csup\u003e-1\u003c/sup\u003e represents N-H and C\u0026thinsp;=\u0026thinsp;O groups of proteins. Peaks from 1400cm\u003csup\u003e-1\u003c/sup\u003e to 1000cm\u003csup\u003e-1\u003c/sup\u003e are due to C-N symmetrical stretching vibrations of amides, aromatic, and aliphatic amines [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. The bands at 750cm\u003csup\u003e-1\u003c/sup\u003e, and 720cm\u003csup\u003e-1\u003c/sup\u003e signifies the presence of alkyl groups. This data confirms that microbial enzymes are responsible for the bio reduction of platinum ions into platinum nanoparticles. X-ray diffraction pattern of biosynthesized platinum nanoparticles between 2θ values of 25 to 80 is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e4\u003c/span\u003eb. the sharp peaks appeared at 2θ values of 38.6\u0026deg;, 45.35\u0026deg;, 61.15\u0026deg;, and 79.25\u0026deg; with matrix progress orientation along (111), (200), (220), and (311) crystal planes. The results are comparable with standard powder diffraction card of JCPDS, platinum file no. 88-2343. Dynamic Light Scattering Particle Size Analyzer (DLS-PSA) (BT-90 Nanoparticle Size Analyzer) also called photon correlation spectroscopy was used for the analysis of biosynthesized platinum nanoparticles [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. The nanoparticle solution is irradiated with electromagnetic beam, which gets scattered due to Brownian motion of the particles and causes their intensity and angle change. Which in turn is adopted to analyze the hydrodynamic diameter of biosynthesized platinum nanoparticles by employing Stokes-Einstein equation. A low concentration aqueous solution of platinum nanoparticles was prepared for analysis. The resulting solution was sonicated for twenty minutes before analysis. The biosynthesized platinum nanoparticles size was found 6.29 to 25.0 nm with a specific surface area of 194.95m\u003csup\u003e2\u003c/sup\u003e/g (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e4\u003c/span\u003ec \u0026amp; d). Scanning electron microscope (SEM) study was carried out on FEI Nova 450 NanoSem. The micrograph elucidates the structure and morphology of the sample. It delivers highly resolved pictures of sample by concentrating on primary, secondary, and backscattered electrons. SEM micrograph of biosynthesized platinum nanoparticles is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e4\u003c/span\u003ee.\u003c/p\u003e\u003cp\u003e(a)\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e(b) (c)\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003col\u003e\u003cspan\u003e\u003cli\u003e\u003cp\u003e(b)\u003c/p\u003e\u003c/li\u003e\u003c/span\u003e\u003c/ol\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e(c) (d)\u003c/p\u003e\u003cp\u003e(e)\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eAntimicrobial activity of biogenic platinum nanoparticles was carried out against four gram positive and four-gram negative bacteria. Gram positive bacterial strains \u003cem\u003eStaphylococcus epidermidis, Staphylococcus aureus, Bacillus subtilis, Bacillus licheniformis\u003c/em\u003e and gram-negative bacterial strains \u003cem\u003eEscherichia coli, Salmonella enteria, Porteus mirabilis, Pseudomonas aeruginosa\u003c/em\u003e were used. Disc diffusion process was used to ascertain the zones of inhibition (mm) of platinum nanoparticles. Penicillin in concentration of 200\u0026micro;g/ml was used as a standard antibiotic against all the eight bacterial strains. Different zones of inhibitions were noticed for microorganisms based on their structure and composition. However anti-microbial agent was found to be more potent for gram negative bacteria (Fig.\u0026nbsp;5a). Broth macro dilution method was used to find conventional microbiological parameters like minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) of the bio fabricated PtNPs (Fig.\u0026nbsp;5b). All experiments were performed in triplicate (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Different reports have been proposed to illustrate the mechanistic aspects of antimicrobial ability of biosynthesized platinum nanoparticles. Due to negative zeta potential of platinum nanoparticles, they may develop chemical interaction with bacterial cell membrane resulting in leakage of cellular contents which is lethal for living cells [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. Platinum nanoparticles can penetrate and interact with cellular organelles and biomolecules and thus disturbing the cellular pathways. Similarly antimicrobial agent may cause creation of reactive oxygen species (ROS) within the cell inducing bactericidal oxidative stress [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. From this it is established that biosynthesized platinum nanoparticles showed good anti-microbial activity against different microbes and can be used for improvement of new anti-microbials generations, nanomedicines, drug delivery systems, and various more.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eDiameter of zones of inhibition (mm) of biosynthesized PtNPs against standard penicillin, MIC, and MLC (\u0026micro;g/ml) by disc diffusion and macro broth dilution methods respectively.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"5\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTested Strains\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePenicillin\u003c/p\u003e\u003cp\u003e(200\u0026micro;g/ml)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003ePtNPs\u003c/p\u003e\u003cp\u003e(200\u0026micro;g/ml)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eMIC\u003c/p\u003e\u003cp\u003e(\u0026micro;g/ml)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eMLC\u003c/p\u003e\u003cp\u003e(\u0026micro;g/ml)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eStaphylococcus epidermidis\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e80\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e140\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eStaphylococcus aureus\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e22\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e10\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e100\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e150\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eBacillus subtilis\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e120\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e160\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eBacillus licheniformis\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e13\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e150\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e200\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eEscherichia coli\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e65\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e100\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003eSalmonella enteria\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e28\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e100\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e180\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003ePorteus mirabilis\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e30\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e25\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e120\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e170\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003e\u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e18\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e60\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e120\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e(a)\u003c/p\u003e\u003cp\u003e(b)\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eFigure 5(a, b)\u003c/strong\u003e\u003cp\u003e(a) Zones of inhibitions of gram-positive and gram-negative bacteria. (b) MIC and MLC of biosynthesized platinum nanoparticles.\u003c/p\u003e\u003c/p\u003e\u003cp\u003eTo examine the antioxidant potential of biosynthesized platinum nanoparticles, DPPH (2,2-diphenyl-1-picrylhydrazyl-hydrate) assay was used. DPPH is purple colored organic compound, undergo a redox reaction, and turned to light yellow in color. On reduction the purple color of DPPH bleached showing antioxidant potential of added sample. The method is simple, sensitive and produces exceptional results for biosynthesized platinum nanoparticles. The antioxidant ability of PtNPs was studied at different time intervals and found that 100\u0026micro;l sample exhibited an excellent radical scavenging activity when compared with Trolox and Butylated hydroxy anisole (BHA) standards (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e6\u003c/span\u003e). The antioxidant activity of DPPH was represented by the percentage DPPH remaining. It showed significant activity (94.74% remaining) after 30 minutes. Antioxidant potential is mostly related to the size of nanoparticles and size depends on the type of microbe used for the synthesis of nanoparticles. However, it is believed that nanoparticles prepared from gram positive bacteria have better antioxidant activity due to more negative zeta potential [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. Microbial methods for synthesis of platinum nanoparticles are superior as microbes have complex and diverse compounds such as enzymes and coenzymes that can only be seen in microbial systems.\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003eBiosynthesized platinum nanoparticles were tested for their catalytic potential. The study was performed for disintegration of methyl orange and methylene blue in industrial wastewater. Methyl orange (MO) is an azo dye and is normally used as an indicator. It is orange red in color and shows maximum absorbance at 460nm. Methylene blue [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e] is a thiazine dye that finds extensive uses. It is deep blue in color and has maximum absorbance at 665nm. Even if sodium borohydride is a strong reducing agent but still is unable to accomplish the reduction of both dyes due to great difference redox potentials of dyes and reducing agent. These reactions are thermally favorable but kinetically unfavorable. The degradation study of methyl orange, and methylene blue with NaBH\u003csub\u003e4\u003c/sub\u003e was carried out in presence and absence of biosynthesized platinum nanoparticles. The blank experiments were performed without platinum nanoparticles exhibited no color change even after 30minutes so there was no degradation of both dyes in the absence of platinum nanoparticles. At the same time, degradation occurred rapidly by adding small amounts of platinum nanoparticles which showed their catalytic potential. The degradation is evident by fading the orange red and deep blue colors of methyl orange and methylene blue as well as decrease in their absorbance at 460nm and 665nm respectively. Spectrophotometer was used to study reaction kinetics (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). The NaBH\u003csub\u003e4\u003c/sub\u003e added is much higher in amount as compared to the dyes, practically its concentration remains constant throughout the course of reaction. So, the kinetics followed by both the reactions were pseudo first order. The concentration of both dyes altered with time. This become clear in (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e7\u003c/span\u003ea, b) which are plots of ln[A] with time for degradation of methyl orange and methylene blue. The process is a redox reaction. The reaction between NaBH\u003csub\u003e4\u003c/sub\u003e and dyes is an electron transfer mechanism in which NaBH\u003csub\u003e4\u003c/sub\u003e is a donor and dye act as accepter. The added platinum nanoparticles have potential between these two and make the transmission of electron easy. Due to the large surface area of platinum nanoparticles, the rate of reduction process increased. Before the onset of reduction, both the dye and borohydride ions (BH\u003csub\u003e4\u003c/sub\u003e \u003csup\u003e\u0026minus;\u003c/sup\u003e) get adsorbed on the surface of catalyst. The borohydride ions release electrons to the surface of catalyst from which dye molecules gain electrons and get reduced. Due to this reason, presence of nano catalyst enhances the rate of reduction process and proved to be an efficient catalyst for the removal of harmful dyes from industrial wastewater [\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. The illegal dumping of waste from industries has rebellious pollutants, which is a worldwide problem with more severity in developing nations.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eDeterioration of methyl orange and methylene blue in presence and absence of biosynthesized platinum nanoparticles along with ln [A] calculated from pseudo first order kinetics.\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"6\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDye\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eTime (min)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eAbsorbance with PtNPs\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003eln[A]\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003eAbsorbance without PtNPs\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003eln[A]\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eMethyl orange\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0\u003c/p\u003e\u003cp\u003e5\u003c/p\u003e\u003cp\u003e10\u003c/p\u003e\u003cp\u003e15\u003c/p\u003e\u003cp\u003e20\u003c/p\u003e\u003cp\u003e25\u003c/p\u003e\u003cp\u003e30\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e2.64\u003c/p\u003e\u003cp\u003e2.58\u003c/p\u003e\u003cp\u003e2.10\u003c/p\u003e\u003cp\u003e1.72\u003c/p\u003e\u003cp\u003e1.29\u003c/p\u003e\u003cp\u003e0.87\u003c/p\u003e\u003cp\u003e0.31\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e0.97\u003c/p\u003e\u003cp\u003e0.94\u003c/p\u003e\u003cp\u003e0.74\u003c/p\u003e\u003cp\u003e0.54\u003c/p\u003e\u003cp\u003e0.25\u003c/p\u003e\u003cp\u003e-0.14\u003c/p\u003e\u003cp\u003e-1.17\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e2.64\u003c/p\u003e\u003cp\u003e2.62\u003c/p\u003e\u003cp\u003e2.55\u003c/p\u003e\u003cp\u003e2.51\u003c/p\u003e\u003cp\u003e2.48\u003c/p\u003e\u003cp\u003e2.44\u003c/p\u003e\u003cp\u003e2.40\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e0.97\u003c/p\u003e\u003cp\u003e0.96\u003c/p\u003e\u003cp\u003e0.93\u003c/p\u003e\u003cp\u003e0.92\u003c/p\u003e\u003cp\u003e0.91\u003c/p\u003e\u003cp\u003e0.89\u003c/p\u003e\u003cp\u003e0.87\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eMethylene blue\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e0\u003c/p\u003e\u003cp\u003e5\u003c/p\u003e\u003cp\u003e10\u003c/p\u003e\u003cp\u003e15\u003c/p\u003e\u003cp\u003e20\u003c/p\u003e\u003cp\u003e25\u003c/p\u003e\u003cp\u003e30\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e2.90\u003c/p\u003e\u003cp\u003e2.81\u003c/p\u003e\u003cp\u003e1.85\u003c/p\u003e\u003cp\u003e1.39\u003c/p\u003e\u003cp\u003e0.81\u003c/p\u003e\u003cp\u003e0.55\u003c/p\u003e\u003cp\u003e0.06\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e1.06\u003c/p\u003e\u003cp\u003e1.03\u003c/p\u003e\u003cp\u003e0.61\u003c/p\u003e\u003cp\u003e0.33\u003c/p\u003e\u003cp\u003e-0.21\u003c/p\u003e\u003cp\u003e-0.59\u003c/p\u003e\u003cp\u003e-2.81\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c5\"\u003e\u003cp\u003e2.90\u003c/p\u003e\u003cp\u003e2.87\u003c/p\u003e\u003cp\u003e2.54\u003c/p\u003e\u003cp\u003e2.01\u003c/p\u003e\u003cp\u003e1.86\u003c/p\u003e\u003cp\u003e1.74\u003c/p\u003e\u003cp\u003e1.57\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c6\"\u003e\u003cp\u003e1.06\u003c/p\u003e\u003cp\u003e1.05\u003c/p\u003e\u003cp\u003e0.93\u003c/p\u003e\u003cp\u003e0.67\u003c/p\u003e\u003cp\u003e0.62\u003c/p\u003e\u003cp\u003e0.55\u003c/p\u003e\u003cp\u003e0.45\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e(a)\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e(b)\u003c/p\u003e\u003cp\u003e\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\u003ch2\u003eMaterials\u003c/h2\u003e\u003cp\u003eAll the chemicals including hexa-chloroplatinic acid H\u003csub\u003e2\u003c/sub\u003ePtCl\u003csub\u003e6\u003c/sub\u003e (99.9%), nutrient broth, agar, DPPH (2,2-diphenyl-1-picrylhydrazyl-hydrate), methyl orange, and methylene blue used in this examination were of analytical grade and procured from Sigma Aldrich Chemicals Germany. Double distilled deionized water was used during this examination. The \u003cem\u003ebacillus cereus\u003c/em\u003e was isolated from dirty and filthy soil and then decontaminated and identified microbiologically. The pure cultures were held in reserve and re-cultured from time to time in nutrient broth agar medium slants. \u003cem\u003eStaphylococcus epidermidis, Staphylococcus aureus, Bacillus subtilis, Bacillus licheniformis, Escherichia coli, Salmonella enteria, Porteus mirabilis\u003c/em\u003e, and \u003cem\u003ePseudomonas aeruginosa\u003c/em\u003e were attained from Mayo Hospital Lahore for antimicrobial activity.\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eInoculum Size Preparation\u003c/h3\u003e\n\u003cp\u003eConical flask (250ml) containing 1.3g nutrient broth per 100ml of buffer solution (pH 5 to 9) was autoclaved at 121\u003csup\u003e\u0026deg;\u003c/sup\u003eC for 30minutes. A loop full of \u003cem\u003eBacillus cereus\u003c/em\u003e was then injected into the autoclaved flask and placed in a shaking incubator at 37\u003csup\u003e◦\u003c/sup\u003eC, 120rpm for 24 hours.\u003c/p\u003e\n\u003ch3\u003eGreen synthesis of Platinum Nanoparticles\u003c/h3\u003e\n\u003cp\u003eProduction of platinum nanoparticles was led by performing all the twenty-six experiments designed through CCD model of RSM (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). The experiments were carried out by mixing 1 to 10ml of platinum salt solution with 1 to 10ml inoculum of \u003cem\u003ebacillus cereus\u003c/em\u003e ranging in pH from 5 to 9 and placed in incubator shaker for specified time of 12 to 72 hours at temperature of 25\u003csup\u003e\u0026deg;\u003c/sup\u003eC to 50\u003csup\u003e\u0026deg;\u003c/sup\u003eC and 120rpm. The response of each experiment was monitored through UV-visible spectrophotometer by measuring absorbance at 290nm. Each experiment was performed in triplicate. After that, optimized factors for maximum biosynthesis of platinum nanoparticles were obtained (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003ea). One to one ratio of platinum salt solution (1millimolar) and inoculum size (\u003cem\u003ebacillus cereus\u003c/em\u003e) maintained at pH eight were mixed and placed in incubator shaker for 58.4hours at 43.2\u003csup\u003e\u0026deg;\u003c/sup\u003eC and 120rpm. The color change from yellow to dark brown was observed (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e3\u003c/span\u003eb). The biosynthesized platinum nanoparticles were separated from reaction mixture through centrifugation for ten minutes at 8000rpm and washed with deionized water for the elimination of active molecules. The nanoparticles were air dried for further studies.\u003c/p\u003e\n\u003ch3\u003eAntimicrobial activity\u003c/h3\u003e\n\u003cp\u003eGram positive bacterial strains \u003cem\u003eStaphylococcus epidermidis, Staphylococcus aureus, Bacillus subtilis, Bacillus licheniformis\u003c/em\u003e and gram-negative bacterial strains \u003cem\u003eEscherichia coli, Salmonella enteria, Porteus mirabilis, Pseudomonas aeruginosa\u003c/em\u003e were used to find the zone of inhibition, minimum inhibitory concentrations (MIC), and minimum lethal concentrations (MLC) of biosynthesized platinum nanoparticles. Kirby-Bauer disc diffusion method [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e] was employed to calculate the zone of inhibitions of the biosynthesized platinum nanoparticles. Each of the bacterial culture was swabbed in the sterilized petri plate with Mueller Hinton agar. Into each petri plate, two discs (6mm), one soaked with standard antibiotic Penicillin (200\u0026micro;g/ml) and second saturated with biosynthesized platinum nanoparticles (200\u0026micro;g/ml), were placed on the uppermost layer. These petri plates were placed in an incubator for 24hours at the temperature of 37\u003csup\u003e\u0026deg;\u003c/sup\u003eC. After that diameter (mm) of zone of inhibition (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e) was measured for each bacterial species (Fig.\u0026nbsp;5a).\u003c/p\u003e\u003cp\u003eThe minimum inhibitory concentration (MIC) represents the lowest concentration of platinum nanoparticles that inhibits the growth (99%) of microbes. Broth macro dilution method was used to find the MIC. To encounter MIC, the Lauria Bertain (LB) broth and two times chronological dilutions of biosynthesized platinum nanoparticles (1600\u0026micro;g/ml to 0.05\u0026micro;g/ml) were used. A test tube with microbe in LB broth acts as positive control while a negative control tube contains LB broth only. The positive and negative controls were also carried out in parallel. All these were placed in an orbital shaker at 37\u003csup\u003e\u0026deg;\u003c/sup\u003eC and 120rpm for 24 hours and after that MIC was determined. For the determination of minimum lethal concentration (MLC), 25\u0026micro;l of each clear tube with no visible bacterial growth was seeded on Muller-Hinton Agar (MHA) plates and placed in an incubator at 37\u003csup\u003e\u0026deg;\u003c/sup\u003eC for 24hours. The MLC represents the lowest concentration of biosynthesized platinum nanoparticles which assassinate 100% of microbial growth so there will be no growth on MHA plates (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Bactericidal effects were represented by MLC while MIC represents the bacteriostatic effects of biosynthesized platinum nanoparticles against the tested microbial strains (Fig.\u0026nbsp;5b).\u003c/p\u003e\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003eAntioxidant Potential\u003c/h2\u003e\u003cp\u003eFor DPPH assay, a standard solution of DPPH in methanol was prepared by dissolving 25mg/l. the absorbance of UV-Visible spectrophotometer was adjusted at 1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02 at 515nm. The solution of bio fabricated platinum nanoparticles was processed in double distilled water by adding 3mg/ml. For antioxidation potential, to the 2.5 ml of methanolic DPPH solution was added 100\u0026micro;l of biosynthesized platinum nanoparticles solution and mixed vigorously. The mixture could stand for 30 minutes in dark. Measure the absorbance after every 5 minutes up to 30 minutes. The scavenging ability was calculated by following expression,\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003e% DPPH remaining = (DPPH)t\u0026thinsp;=\u0026thinsp;t/ (DPPH)t\u0026thinsp;=\u0026thinsp;0 \u0026times; 100\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003ewhere (DPPH)t\u0026thinsp;=\u0026thinsp;t was absorbance of DPPH solution at a specific time and (DPPH)t\u0026thinsp;=\u0026thinsp;0 showed absorbance of DPPH solution at the initial time.\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eCatalytic Degradation of dyes\u003c/h3\u003e\n\u003cp\u003eCatalytic disintegration of methyl orange (MO) and methylene blue [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e] through NaBH\u003csub\u003e4\u003c/sub\u003e was investigated when biosynthesized platinum nanoparticles act as catalyst. The reaction was performed by mixing 2ml aqueous solution (0.01\u0026times;10\u003csup\u003e\u0026minus;\u0026thinsp;2\u003c/sup\u003e) of each of the dyes with 0.5ml (0.006M) solution of sodium borohydride followed by 0.5ml (0.05mg/ml) of biosynthesized platinum nanoparticles. The progress of two reactions and change in concentration of methyl orange and methylene blue with time was measured via UV-visible spectrophotometer at 460nm and 665nm respectively. Figure out the change in absorbance of each solution after every 5 minutes delay for 30 minutes. Absorbance is directly proportional to concentration, so a decrease in concentration results a decrease in absorption (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Two blank reactions for each dye without biosynthesized platinum nanoparticles were also performed. For representation of reduction of methyl orange and methylene blue in presence and absence of platinum nanoparticles plots of ln[A] along time are shown in (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e7\u003c/span\u003ea, b).\u003c/p\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eResultantly, \u003cem\u003ebacillus cereus\u003c/em\u003e was utilized for the biogenic synthesis of platinum nanoparticles. Central composite design (CCD) of response surface methodology (RSM) was used for the optimization of reaction conditions such as metal salt concentration, inoculum size, temperature, time, and pH. On the basis of ANOVA results for quadratic model, three dimensional responses, and perturbation plot optimized conditions for small sized, high surface area, biosynthesized platinum nanoparticles are, platinum salt solution (1mmol)\u0026thinsp;=\u0026thinsp;3.0ml, inoculum size (\u003cem\u003ebacillus cereus\u003c/em\u003e)\u0026thinsp;=\u0026thinsp;3ml, temperature\u0026thinsp;=\u0026thinsp;43.2\u003csup\u003e◦\u003c/sup\u003eC, time\u0026thinsp;=\u0026thinsp;58.4hours, and pH\u0026thinsp;=\u0026thinsp;8.0. By following these set of optimized conditions, the biosynthesized platinum nanoparticles were of size 6.29 to 25.0nm with 194.95m\u003csup\u003e2\u003c/sup\u003e/g surface area. The CCD of RSM detected as a vital tool to explain the distinct and combined properties of various tentative data points and their optimization for improved biosynthesis of platinum nanoparticles in a cost effective and time effective method. Uv-visible spectrophotometer, FTIR spectroscopy, DLS-particle size analyzer, XRD, and SEM were used for characterization of biosynthesized platinum nanoparticles. For antibacterial activity, four gram-positive bacterial strains \u003cem\u003eStaphylococcus epidermidis, Staphylococcus aureus, Bacillus subtilis, Bacillus licheniformis\u003c/em\u003e and four gram-negative bacterial strains \u003cem\u003eEscherichia coli, Salmonella enteria, Porteus mirabilis, Pseudomonas aeruginosa\u003c/em\u003e were used with standard antibiotic Penicillin to calculate the zones of inhibitions by disc diffusion method and MIC and MLC by broth macro dilution method. The biosynthesized platinum nanoparticles were also evaluated for their antioxidant potential through DPPH (2,2-diphenyl-1-picrylhydrazyl) assay. Catalytic role of biosynthesized platinum nanoparticles was assessed for disintegration of methyl orange and methylene blue with time and give brilliant results. It is first ever report in which platinum nanoparticles were synthesized through \u003cem\u003ebacillus cereus\u003c/em\u003e and optimization of experimental conditions were carried out through CCD of RSM. The biosynthesized platinum nanoparticles were also studied for their biomedical and environmental applications.\u003c/p\u003e\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003eDisclosure Declaration\u003c/h2\u003e\u003cp\u003eThe authors have no conflict of interests that can impact the reported research work\u003c/p\u003e\u003c/div\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eCCD\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Central Composite Design\u003c/p\u003e\n\u003cp\u003eRSM\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Response Surface Methodology\u003c/p\u003e\n\u003cp\u003eNPs\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;nanoparticles\u003c/p\u003e\n\u003cp\u003ePtNPs\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;platinum nanoparticles\u003c/p\u003e\n\u003cp\u003eFTIR\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Fourier transform infrared spectroscopy\u003c/p\u003e\n\u003cp\u003eXRD\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;x-ray diffraction\u003c/p\u003e\n\u003cp\u003eDLS\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Dynamic light scattering\u003c/p\u003e\n\u003cp\u003eSEM\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Scanning Electron Microscopy\u003c/p\u003e\n\u003cp\u003eMIC\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;Minimum Inhibitory Concentration\u003c/p\u003e\n\u003cp\u003eMLC\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Minimum Lethal Concentration\u003c/p\u003e\n\u003cp\u003eBHA Butylated hydroxy anisole\u003c/p\u003e\n\u003cp\u003eDPPH 2,2-diphenyl-1-picrylhydrazyl\u003c/p\u003e\n\u003cp\u003eMO\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Methyl orange\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/p\u003e\n\u003cp\u003eMB\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;Methylene blue\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003ch2\u003eEthical Approval\u003c/h2\u003e\u003cp\u003eThis item does not comprise any studies with humans or animals made by any of the authors.\u003c/p\u003e\u003c/p\u003e\u003ch2\u003eFunding\u003c/h2\u003e\u003cp\u003eDeclaration\u003c/p\u003e\u003cp\u003eThis research received no external funding or financial support.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eS I performed experiments and write the manuscript. Z A provide assistance in experiments. S I and M M prepare all the figures.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eIqbal, J., et al. (2025). \u003cem\u003eMicrobial Synthesis of Metal Nanoparticles for Nanomedicinal and Catalytic Applications, in Expanding Nanobiotechnology: Applications and Commercialization\u003c/em\u003e (pp. 88\u0026ndash;124). CRC.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eJameel, M. S., Aziz, A. A., \u0026amp; Dheyab, M. A. (2020). Green synthesis: Proposed mechanism and factors influencing the synthesis of platinum nanoparticles. \u003cem\u003eGreen processing and synthesis\u003c/em\u003e, \u003cem\u003e9\u003c/em\u003e(1), 386\u0026ndash;398.\u003c/span\u003e\u003c/li\u003e\u003cli\u003e\u003cspan\u003eKirubakaran, D., et al. 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Reliability of the Kirby-Bauer disc diffusion method for detecting methicillin-resistant strains of Staphylococcus aureus. \u003cem\u003eApplied microbiology\u003c/em\u003e, \u003cem\u003e24\u003c/em\u003e(2), 240\u0026ndash;247.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Platinum nanoparticles, Bacillus Cereus, RSM, CCD, Antimicrobial activity, Antioxidant, Catalytic potential","lastPublishedDoi":"10.21203/rs.3.rs-6922293/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6922293/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eTo progress one step biosynthetic process, reciprocal effect of reaction variables like concentration of platinum salt solution, inoculum size, temperature, time, and pH was considered by making use of central composite design of response surface methodology. The optimized set of reaction conditions were platinum salt solution (1mmol)\u0026thinsp;=\u0026thinsp;3.0ml, inoculum size (\u003cem\u003ebacillus cereus\u003c/em\u003e)\u0026thinsp;=\u0026thinsp;3ml, temperature\u0026thinsp;=\u0026thinsp;43.2\u003csup\u003e◦\u003c/sup\u003eC, time\u0026thinsp;=\u0026thinsp;58.4hours, and pH\u0026thinsp;=\u0026thinsp;8.0. The biogenic platinum nanoparticles were categorized by UV-vis spectroscopy, FTIR, DLS-PSA, XRD and SEM. The UV-visible spectra showed a surface plasmon response at 290nm. FTIR spectra revealed the existence of enzymatic proteins responsible for fabrication and stabilization of platinum nanoparticles. DLS-PSA, XRD and SEM confirmed the formation of isotropic and spherical platinum nanoparticles of size 6.29 to 25.0 nm with a specific surface area of 194.95m\u003csup\u003e2\u003c/sup\u003e/g. Moreover, gram positive bacterial strains \u003cem\u003eStaphylococcus epidermidis, Staphylococcus aureus, Bacillus subtilis, Bacillus licheniformis\u003c/em\u003e and gram-negative bacterial strains \u003cem\u003eEscherichia coli, Salmonella enteria, Porteus mirabilis, Pseudomonas aeruginosa\u003c/em\u003e were tested for antibacterial activity by employing disc diffusion and broth macro dilution methods. DPPH (2,2-diphenyl-1-picrylhydrazyl) assay was carried out to monitor the free radical scavenging ability of biogenic platinum nanoparticles. Furthermore, the degradation rate of methyl orange and methylene blue in presence pf platinum nanoparticles as catalyst was investigated successfully. The bio fabricated platinum nanoparticles found applications in biomedical and environmental remediation.\u003c/p\u003e","manuscriptTitle":"Central Composite Design for Optimizing the Biogenic fabrication of Platinum Nanoparticles Using bacillus cereus and TheirAntimicrobial, Antioxidant, and catalytic Potential","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-10 18:09:10","doi":"10.21203/rs.3.rs-6922293/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"9a355a71-770a-4f6d-b0ea-4b7add9201e0","owner":[],"postedDate":"July 10th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-08-07T07:40:31+00:00","versionOfRecord":[],"versionCreatedAt":"2025-07-10 18:09:10","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6922293","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6922293","identity":"rs-6922293","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}