β-nicotinamide mononucleotide production in Vibrio natriegens: a preliminary study

β-nicotinamide mononucleotide production in Vibrio natriegens: a preliminary study | bioRxiv /* */ /* */ <!-- <!-- /*! * yepnope1.5.4 * (c) WTFPL, GPLv2 */ (function(a,b,c){function d(a){return"[object Function]"==o.call(a)}function e(a){return"string"==typeof a}function f(){}function g(a){return!a||"loaded"==a||"complete"==a||"uninitialized"==a}function h(){var a=p.shift();q=1,a?a.t?m(function(){("c"==a.t?B.injectCss:B.injectJs)(a.s,0,a.a,a.x,a.e,1)},0):(a(),h()):q=0}function i(a,c,d,e,f,i,j){function k(b){if(!o&&g(l.readyState)&&(u.r=o=1,!q&&h(),l.onload=l.onreadystatechange=null,b)){"img"!=a&&m(function(){t.removeChild(l)},50);for(var d in y[c])y[c].hasOwnProperty(d)&&y[c][d].onload()}}var j=j||B.errorTimeout,l=b.createElement(a),o=0,r=0,u={t:d,s:c,e:f,a:i,x:j};1===y[c]&&(r=1,y[c]=[]),"object"==a?l.data=c:(l.src=c,l.type=a),l.width=l.height="0",l.onerror=l.onload=l.onreadystatechange=function(){k.call(this,r)},p.splice(e,0,u),"img"!=a&&(r||2===y[c]?(t.insertBefore(l,s?null:n),m(k,j)):y[c].push(l))}function j(a,b,c,d,f){return q=0,b=b||"j",e(a)?i("c"==b?v:u,a,b,this.i++,c,d,f):(p.splice(this.i++,0,a),1==p.length&&h()),this}function k(){var a=B;return a.loader={load:j,i:0},a}var l=b.documentElement,m=a.setTimeout,n=b.getElementsByTagName("script")[0],o={}.toString,p=[],q=0,r="MozAppearance"in l.style,s=r&&!!b.createRange().compareNode,t=s?l:n.parentNode,l=a.opera&&"[object Opera]"==o.call(a.opera),l=!!b.attachEvent&&!l,u=r?"object":l?"script":"img",v=l?"script":u,w=Array.isArray||function(a){return"[object Array]"==o.call(a)},x=[],y={},z={timeout:function(a,b){return b.length&&(a.timeout=b[0]),a}},A,B;B=function(a){function b(a){var a=a.split("!"),b=x.length,c=a.pop(),d=a.length,c={url:c,origUrl:c,prefixes:a},e,f,g;for(f=0;f<d;f++)g=a[f].split("="),(e=z[g.shift()])&&(c=e(c,g));for(f=0;f<b;f++)c=x[f](c);return c}function g(a,e,f,g,h){var i=b(a),j=i.autoCallback;i.url.split(".").pop().split("?").shift(),i.bypass||(e&&(e=d(e)?e:e[a]||e[g]||e[a.split("/").pop().split("?")[0]]),i.instead?i.instead(a,e,f,g,h):(y[i.url]?i.noexec=!0:y[i.url]=1,f.load(i.url,i.forceCSS||!i.forceJS&&"css"==i.url.split(".").pop().split("?").shift()?"c":c,i.noexec,i.attrs,i.timeout),(d(e)||d(j))&&f.load(function(){k(),e&&e(i.origUrl,h,g),j&&j(i.origUrl,h,g),y[i.url]=2})))}function h(a,b){function c(a,c){if(a){if(e(a))c||(j=function(){var a=[].slice.call(arguments);k.apply(this,a),l()}),g(a,j,b,0,h);else if(Object(a)===a)for(n in m=function(){var b=0,c;for(c in a)a.hasOwnProperty(c)&&b++;return b}(),a)a.hasOwnProperty(n)&&(!c&&!--m&&(d(j)?j=function(){var a=[].slice.call(arguments);k.apply(this,a),l()}:j[n]=function(a){return function(){var b=[].slice.call(arguments);a&&a.apply(this,b),l()}}(k[n])),g(a[n],j,b,n,h))}else!c&&l()}var h=!!a.test,i=a.load||a.both,j=a.callback||f,k=j,l=a.complete||f,m,n;c(h?a.yep:a.nope,!!i),i&&c(i)}var i,j,l=this.yepnope.loader;if(e(a))g(a,0,l,0);else if(w(a))for(i=0;i (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];var j=d.createElement(s);var dl=l!='dataLayer'?'&l='+l:'';j.src='//www.googletagmanager.com/gtm.js?id='+i+dl;j.type='text/javascript';j.async=true;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-M677548'); Skip to main content Home About Submit ALERTS / RSS Search for this keyword Advanced Search New Results β-nicotinamide mononucleotide production in Vibrio natriegens : a preliminary study Ly Huong Tran , Vu Thao Phuong Tran , Huy Tuan Dat Pham , Giang Tra Nguyen , Tuoi Thi Nghiem , Anh Duc Pham , Nga Quynh Pham , Thuy Thi Le , Ngoc Lan Nguyen , Tran Nhat Minh Dang , Thinh Huy Tran , Van Khanh Tran , Hoa Quang Le doi: https://doi.org/10.1101/2025.03.17.643733 Ly Huong Tran 1 School of Chemistry and Life Sciences, Hanoi University of Science and Technology , 1st Dai Co Viet, Hanoi, Vietnam Find this author on Google Scholar Find this author on PubMed Search for this author on this site Vu Thao Phuong Tran 2 High school of Education Sciences, University of Education, Vietnam National University , 144 Xuan Thuy, Hanoi, Vietnam Find this author on Google Scholar Find this author on PubMed Search for this author on this site Huy Tuan Dat Pham 2 High school of Education Sciences, University of Education, Vietnam National University , 144 Xuan Thuy, Hanoi, Vietnam Find this author on Google Scholar Find this author on PubMed Search for this author on this site Giang Tra Nguyen 1 School of Chemistry and Life Sciences, Hanoi University of Science and Technology , 1st Dai Co Viet, Hanoi, Vietnam Find this author on Google Scholar Find this author on PubMed Search for this author on this site Tuoi Thi Nghiem 1 School of Chemistry and Life Sciences, Hanoi University of Science and Technology , 1st Dai Co Viet, Hanoi, Vietnam Find this author on Google Scholar Find this author on PubMed Search for this author on this site Anh Duc Pham 1 School of Chemistry and Life Sciences, Hanoi University of Science and Technology , 1st Dai Co Viet, Hanoi, Vietnam Find this author on Google Scholar Find this author on PubMed Search for this author on this site Nga Quynh Pham 1 School of Chemistry and Life Sciences, Hanoi University of Science and Technology , 1st Dai Co Viet, Hanoi, Vietnam Find this author on Google Scholar Find this author on PubMed Search for this author on this site Thuy Thi Le 3 National Institute for Food Control , 65 Pham Than Duat Street, Hanoi, Vietnam Find this author on Google Scholar Find this author on PubMed Search for this author on this site Ngoc Lan Nguyen 5 Center for Gene and Protein Research, Hanoi Medical University , 1st Ton That Tung Street, Hanoi, 11521, Vietnam Find this author on Google Scholar Find this author on PubMed Search for this author on this site Tran Nhat Minh Dang 4 Hanoi Medical University , 1st Ton That Tung Street, Hanoi, 11521, Vietnam Find this author on Google Scholar Find this author on PubMed Search for this author on this site Thinh Huy Tran 4 Hanoi Medical University , 1st Ton That Tung Street, Hanoi, 11521, Vietnam Find this author on Google Scholar Find this author on PubMed Search for this author on this site Van Khanh Tran 5 Center for Gene and Protein Research, Hanoi Medical University , 1st Ton That Tung Street, Hanoi, 11521, Vietnam Find this author on Google Scholar Find this author on PubMed Search for this author on this site Hoa Quang Le 1 School of Chemistry and Life Sciences, Hanoi University of Science and Technology , 1st Dai Co Viet, Hanoi, Vietnam Find this author on Google Scholar Find this author on PubMed Search for this author on this site For correspondence: hoa.lequang{at}hust.edu.vn Abstract Full Text Info/History Metrics Supplementary material Preview PDF ABSTRACT Background Nicotinamide mononucleotide (NMN), is a promising nutraceutical attracting much attention for its pharmacological and anti-aging efficacies. However, NMN-containing commercial products are very high-priced due to the lack of efficient and facile methods for industrial-scale production. To date, various metabolic engineering strategies have been successfully applied to produce NMN in Escherichia coli . Recently, Vibrio natriegens has become a promising host in the bioindustry thanks to its rapid growth and capabilities of broad substrate utilization. This study aims to evaluate the NMN biosynthesis capability of V. natriegens . Methods Firstly, a mutant V. natriegens strain (Δdns::araC-T7RNAP-Kan R ΔpncC::FtnadE-Sm R ΔnadR) was generated via multiplex genome editing by natural transformation (MuGENT). Nampt genes encoding nicotinamide phosphoribosyltransferase from Chitinophaga pinensis, Sphingopyxis sp . C-1, Haemophilus ducreyi, and Vibrio phage KVP40 were codon-optimized and cloned into pACYCDuet™-1 under the control of T7 promoter. The recombinant plasmids were electroporated into the mutant strain. The expression of recombinant NAMPTs in V. natriegens was evaluated by SDS-PAGE analysis and the intracellular NMN concentrations were quantified by HPLC. Results After two rounds of MuGENT, V. natriegens V54-33 strain (Δdns::araC-T7RNAP-Kan R ΔpncC::FtnadE-Sm R ΔnadR) was successfully generated. SDS-PAGE analysis demonstrated that all NAMPTs were strongly expressed in the V54-33 strain. HPLC analysis revealed that the highest intracellular NMN concentration was obtained with NAMPT from Chitinophaga pinensis (44.5 μM), followed by NAMPT from Vibrio phage KVP40 (23.3 μM). Conclusion This study demonstrated the feasibility of NMN biosynthesis in V. natriegens . INTRODUCTION β-nicotinamide mononucleotide (NMN) is a precursor of NAD + , an essential coenzyme in various metabolic processes. NAD + is involved in intracellular signaling pathways, regulating mitochondrial functions and other biochemical processes. 1 In living organisms, intracellular NAD + levels are directly influenced by its precursor, NMN. Recent studies reported various potential medical applications of NMN, particularly in anti-aging, cognitive-enhancing, and the treatment of severe chronic diseases such as Alzheimer’s and diabetes. 2 , 3 There are three major ways for NMN production: chemical synthesis, enzymatic methods and fermentation methods using genetically modified microbial strains. 4 The chemical synthesis pathway offers the advantage of low production costs; however, the resulting isomeric compounds may exhibit toxicity or lack of biological activity. The enzymatic methods enhance selectivity and reduce toxicity, but the high costs of enzymes and phosphate sources such as ATP remain a major obstacle. Recent studies indicated that the fermentation approach using metabolically engineered microbial strains represents a promising solution to overcome the limitations of the latter methods. The most effective strategy for NMN biosynthesis in microbial cells primarily relies on: nicotinamide (NAM) supplement as a substrate, activation of pentose phosphate pathway to generate the precursor PRPP from carbon sources, overexpression of nicotinamide phosphoribosyltransferase (NAMPT) to catalyze the reaction between NAM and PRPP to produce NMN, and the expression of NMN transporter. 5 , 6 Among these factors, NAMPT is the key enzyme that dictates NMN production efficiency. 5 – 7 In a pioneer study, Marinescu et al . evaluated NAMPT from various sources and found that NAMPT from Haemophilus ducreyi , when expressed in E. coli BL21(DE3)pLysS, gave the highest intracellular NMN concentration (15.42 mg/L). 7 Subsequently, Black et al . has combined the overexpression of both NAMPT and NMN synthase from Francisella tularensis (FtNadE) with the deletion of pncC and nadR , two genes involved in the catabolism of NMN, in E. coli , and achieved an intracellular NMN concentration of 1.5 mM. 8 In another study, NAMPTs from ten organisms have been tested for the activity when expressed in E. coli and the results revealed that NAMPTs from Chitinophaga pinensis and Sphingopyxis sp. C-1 displayed significantly higher activities compared to NAMPTs derived from other sources. 5 Similarly, Huang et al . performed a screening of eight NAMPTs from various sources and found that NAMPT from Vibrio phage KVP40 could biosynthesize 81.3 mg/L NMN intracellularly. 6 Recently, Vibrio natriegens has emerged as a host for industrial biotechnology due to its extremely rapid growth rate with a reported doubling time of less than 10 min, non-pathogenic nature, genetic tractability, and ability to utilize a variety of carbon sources. 9 In numerous studies, V. natriegens was used as an alternative expression system for recombinant protein production, 10 and high-value metabolites such as pyruvate, 11 2,3-butanediol. 12 However, the application of V. natriegens in NMN production remains unexplored. Therefore, the present study aimed to evaluate the feasibility of NMN biosynthesis in V. natriegens . MATERIALS AND METHODS Materials Vibrio natriegens strain TND1964, kindly offered by Prof. Ankur Dalia from Indiana University, Bloomington, USA and Escherichia coli DH5α (Thermo Scientific™, cat. number EC0112) were used as hosts for expression and cloning, respectively. Oligonucleotides were synthesized by Macrogen (Korea). Expression vector used in this study was pACYCDuet™-1 (Novagen, cat. number 71147-3). All other reagents were obtained from Thermo Scientific, New England Biolabs, Merck, Qiagen, and Bio Basic, unless otherwise specified. Generation of the recombinant Vibrio natriegens V54-33 strain Vibrio natriegens V54-33 strain (Δdns::araC-T7RNAP-Kan R ΔpncC::FtnadE-Sm R ΔnadR) was generated using multiplex genome editing by natural transformation (MuGENT) method. 13 Details were presented in the Supplement I. 14 Construction of the pACYC-nampt plasmids The Nampt genes encoding nicotinamide phosphoribosyltransferase (NAMPT) from C. pinensis, Sphingopyxis sp. C-1, H. ducreyi , and Vibrio phage KVP40, were codon-optimized for expression in V. natriegens and synthesized by Genscript. These genes were cloned into the pACYCDuet™-1 ( Novagen, cat. number 71147 ) under the control of T7 promoter-1 (Supplement II). 14 Electroporation into Vibrio natriegens V54-33 strain The preparation of V. natriegens electrocompetent cells and the electroporation of DNA plasmids were carried out following the procedures described by Weinstock et al . 15 Transformants were selected on LBv2 agar plates supplemented with Chloramphenicol (25 μg/mL) (Bio Basic, cat. number CB0118). LBv2 is LB-Miller (Bio Basic, cat. number SD7003) supplemented with v2 salts (204 mM NaCl (Bio Basic, cat. number DB0483), 4.2 mM KCl (Bio Basic, cat. number PB0440, and 23.14 mM MgCl 2 (Bio Basic, cat. number MB0328)). Shake Flask Fermentation A single colony was inoculated into 2 mL of LBv2 medium supplemented with Chloramphenicol (25 μg/mL) (Bio Basic, cat. number CB0118) and cultured at 30 °C, 200 rpm overnight. Subsequently, 1% (v/v) of the overnight culture was reinoculated into 10 mL of fresh LBv2 at 30 °C and 200 rpm. Induction was carried out by adding 2 g/L arabinose (Bio Basic, cat. number AB0071L) when OD 600 reached 0.6−0.8. The cultures were cultivated until OD 600 reached 5-6 and the cultured cells, corresponding to 5 mL cultures, were collected for SDS-PAGE analysis. The remaining cultures were centrifuged at 3000 g for 5 min at 4 o C. After discarding the supernatant, the cells were washed in 20 mL modified M9 medium 16 and resuspended in 5 mL modified M9 medium supplemented with Chloramphenicol (25 μg/mL) (Bio Basic, cat. number CB0118), 1 mM nicotinamide (Sigma, cat. number N0636) and 1 mM nicotinic acid (Sigma, cat. number N0765). The cells were cultured at 30 o C and 200 rpm for 5 h and cultured cells, corresponding to 1 OD 600 , were collected, washed with 1 mL of water and stored at -80 o C until NMN quantification. NMN quantification by HPLC NMN quantification was carried out according to the method described by Vu et al . 17 Briefly, cells were resuspended in 500 μL of water and lysed by sonication on ice by using VC 130 System (Sonics & Materials, Inc, cat. number VC130). The amplitude was set at 80% and on/off pulse of 10 s duration each was given for 1 min. Subsequently, 100 μL of trichloroacetic acid 25% (Bio Basic, cat. number TB0968) was added to inactivate the enzymes in the lysate. The obtained mixture was clarified by centrifugation at 15000 g for 15 min at 4 o C and the supernatant was used for HPLC analysis. Supernatants were run on a Shimadzu LC-20A high-performance liquid chromatography (HPLC) system with a Reliant TM C18 chromatography column (150 mm x 4.6 mm, 5 µm) (Waters, cat. number 186007282). Mobile phase used for separation was 10 mM phosphate buffer pH 3: methanol (90:10) (Supelco, cat. number 106007), at the flow rate of 1.0 mL/min. The injection volume was 20 μL, and individual peak areas were determined using a photo diode array at 261 nm. RESULTS AND DISCUSSION In our present study, a metabolic engineering approach ( Figure 1 ) combining the insertion of key enzymes in the NMN biosynthetic pathway (NMN synthase from Francisella tularensis (FtNadE) and the most promising NAMPTs from previous studies) and the inactivation of endogenous NMN-degrading enzymes (NMN amidohydrolase, encoded by pncC , and NMN adenylyltransferase, encoded by nadR ) was used to evaluate the NMN production capability in V. natriegens . To this end, a mutant strain, called V54-33, has been successfully generated by two rounds of MuGENT introducing araC-T7 RNA polymerase construct and FtNadE under the control of T7 promoter, respectively, while deleting pncC and nadR genes (Supplement I). 14 This allowed a tight induction of NAMPTs and FtNadE by arabinose, and thereby provided a controlled regulation for NMN biosynthesis. Of note, our previous attempts using T7 IPTG-induced system were unsuccessful because of the growth inhibition in transformants (possibly due to the leaky expression of T7lac promoter) (data not shown). Download figure Open in new tab Figure 1. Biosynthesis routes of β-nicotinamide mononucleotide used in this study (modified from 8). NMN: nicotinamide mononucleotide, NaMN: nicotinic acid mononucleotide, NAD+: nicotinamide adenine dinucleotide, NAM: nicotinamide, NAMPT: nicotinamide phosphoribosyltransferase, PncC: NMN amidohydrolase, FtNadE: NMN synthetase from Francisella tularensis , NadR: NMN adenylyltransferase. Subsequently, four plasmids overexpressing NAMPTs from C. pinensis (CP), Sphingopyxis sp. C-1 (SSC), H. ducreyi (HD) and Vibrio phage KVP40 (VP) under the control of T7 promoter, were successfully generated (Supplement II). 14 These four enzymes have been reported as the most efficient NAMPTs in separated studies. 5 – 7 In the next step, the strain V54-33 has been transformed with these plasmids harboring NAMPTs. SDS-PAGE analysis ( Figure 2 ) clearly showed that all recombinant strains carrying NAMPT plasmids displayed an intense band at the expected size of 56 kDa after induction with arabinose, while the original V54-33 strain did not possess this band. These results demonstrated that NAMPTs from CP, HD, SSC and VP were successfully expressed in V. natriegens . Download figure Open in new tab Figure 2. SDS-PAGE analysis of the expression of recombinant NAMPTs in V54-33 strain. L: GangNam-STAIN™ Prestained Protein Ladder. CP: lysate of V54-33 strain overexpressing NAMPT from Chitinophaga pinensis . HD: lysate of V54-33 strain overexpressing NAMPT from Haemophilus ducreyi . SSC: lysate of V54-33 strain overexpressing NAMPT from Sphingopyxis sp. C-1. VP: lysate of V54-33 strain overexpressing NAMPT from Vibrio phage KVP40. CT: lysate of V54-33 strain. These recombinant strains were then transferred into modified M9 medium to assess NMN biosynthesis. HPLC quantification revealed that three out of four transformants (CP, HD, and VP) were capable of intracellular NMN accumulation, while SSC transformants and the V54-33 strain exhibited NMN levels below the limit of detection (LOD) of the method ( Figure 3 , Supplement III). 14 Among them, CP transformants exhibited the highest intracellular NMN titers (44.5 μM), whereas HD transformants showed significantly lower NMN accumulation (12.1 μM, p = 0.0111) ( Figure 3 , Supplement III). 14 The NMN concentration of VP transformants was intermediate (23.3 μM) (Supplement III), 14 which aligns with prior research by Huang et al ., showing that NAMPT from VP was more efficient in NMN accumulation than NAMPT from HD. 6 When considering the absolute NMN titers reported in E. coli , NMN accumulation in CP transformants was comparable to that reported by Marinescu et al . (15.42 mg/L), 7 but significantly lower than those reported by Black et al . (1.5 mM) 8 and Huang et al . (81.3 mg/L). 6 These disparities could be explained by the difference in expression systems: (i) FtNadE was expressed on a multiple-copy plasmid in E. coli in the study by Black et al . 8 while this gene was incorporated into the chromosome of V. natriegens as a single-copy in our study; (ii) ushA encoding 5′-phosphatase capable of dephosphorylation of NMN to nicotinamide riboside and purR regulating the synthesis of the precursor phosphoribosyl pyrophosphate (PRPP) were knocked out in the study of Huang et al . 6 whereas these genes have not been inactivated in the present study. Download figure Open in new tab Figure 3. Quantification of intracellular NMN concentration of V54-33 strains harboring pACYC-Nampt plasmids. CP: V54-33 strain overexpressing NAMPT from Chitinophaga pinensis . HD: V54-33 strain overexpressing NAMPT from Haemophilus ducreyi . SSC: V54-33 strain overexpressing NAMPT from Sphingopyxis sp. C-1. VP: V54-33 strain overexpressing NAMPT from Vibrio phage KVP40. Control: V54-33 strain. <LOD: concentration lower than limit of detection of HPLC method. Data are expressed as mean ± S.D. (error bars). Statistical analysis of differences between groups was performed by unpaired t-test (p < 0.05). In summary, this study demonstrated the feasibility of NMN biosynthesis in V. natriegens . Further research focusing on the activation of pentose phosphate pathway and the transporters of NMN and NAM is underway to improve NMN production in this promising host. Competing interests No competing interests were disclosed. Grant information Research reported in this publication was supported by Hanoi University of Science and Technology [Grant number: T2022-TĐ-002] The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. ETHICS AND CONSENT Ethical approval and consent were not required. DATA AVAILABILITY Underlying data Dataset from NCBI Sequence Read Archive: Sequencing results for bioproject: “β-nicotinamide mononucleotide production in Vibrio natriegens: a preliminary study” Accession number PRJNA1231393; https://www.ncbi.nlm.nih.gov/sra/?term=PRJNA1231393 Data are available under the terms of the Creative Commons Zero “No rights reserved” data waiver (CC0 1.0 Public domain dedication). Extended data Figshare: Supplement for “β-nicotinamide mononucleotide production in Vibrio natriegens: a preliminary study”. 14 https://figshare.com/articles/dataset/Supplement_for_-nicotinamide_mononucleotide_production_in_Vibrio_natriegens_a_preliminary_study_/28547603/1 This project contains the following extended data: Supplement I: Generation of Vibrio natriegens strain V54-33 (Δdns::araC-T7RNAP-Kan R ΔpncC::FtnadE-Sm R ΔnadR) .pdf Supplement II: Generation of pACYCDuet-Nampt plasmids.pdf Supplement III: Quantification of intracellular NMN concentration by HPLC.pdf Data are available under the terms of the Creative Commons Zero “No rights reserved” data waiver (CC0 1.0 Public domain dedication). REFERENCES 1. ↵ Dölle C , Rack JGM , Ziegler M. NAD and ADP-ribose metabolism in mitochondria . FEBS Journal . 2013 ; 280 ( 15 ). doi: 10.1111/febs.12304 OpenUrl CrossRef PubMed 2. ↵ Popescu RG , Dinischiotu A , Soare T , Vlase E , Marinescu GC . Nicotinamide Mononucleotide (NMN) Works in Type 2 Diabetes through Unexpected Effects in Adipose Tissue, Not by Mitochondrial Biogenesis . Int J Mol Sci . 2024 ; 25 ( 5 ). doi: 10.3390/ijms25052594 OpenUrl CrossRef 3. ↵ Campbell JM . Supplementation with NAD+ and Its Precursors to Prevent Cognitive Decline across Disease Contexts . Nutrients . 2022 ; 14 ( 15 ). doi: 10.3390/nu14153231 OpenUrl CrossRef 4. ↵ Luo S , Zhao J , Zheng Y , Chen T , Wang Z. Biosynthesis of Nicotinamide Mononucleotide: Current Metabolic Engineering Strategies, Challenges, and Prospects . Fermentation . 2023 ; 9 ( 7 ). doi: 10.3390/fermentation9070594 OpenUrl CrossRef 5. ↵ Shoji S , Yamaji T , Makino H , Ishii J , Kondo A. Metabolic design for selective production of nicotinamide mononucleotide from glucose and nicotinamide . Metab Eng . 2021 ; 65 . doi: 10.1016/j.ymben.2020.11.008 OpenUrl CrossRef 6. ↵ Huang Z , Li N , Yu S , Zhang W , Zhang T , Zhou J. Systematic Engineering of Escherichia coli for Efficient Production of Nicotinamide Mononucleotide from Nicotinamide . ACS Synth Biol . 2022 ; 11 ( 9 ). doi: 10.1021/acssynbio.2c00100 OpenUrl CrossRef 7. ↵ Marinescu GC , Popescu RG , Stoian G , Dinischiotu A. β-nicotinamide mononucleotide (NMN) production in Escherichia coli . Sci Rep . 2018 ; 8 ( 1 ). doi: 10.1038/s41598-018-30792-0 OpenUrl CrossRef 8. ↵ Black WB , Aspacio D , Bever D , King E , Zhang L , Li H. Metabolic engineering of Escherichia coli for optimized biosynthesis of nicotinamide mononucleotide, a noncanonical redox cofactor . Microb Cell Fact . 2020 ; 19 ( 1 ). doi: 10.1186/s12934-020-01415-z OpenUrl CrossRef PubMed 9. ↵ Thoma F , Blombach B. Metabolic engineering of Vibrio natriegens . Essays Biochem . 2021 ; 65 ( 2 ). doi: 10.1042/EBC20200135 OpenUrl CrossRef PubMed 10. ↵ Smith M , Hernández JS , Messing S , et al. Producing recombinant proteins in Vibrio natriegens . Microb Cell Fact . 2024 ; 23 ( 1 ). doi: 10.1186/s12934-024-02455-5 OpenUrl CrossRef 11. ↵ Wu F , Wang S , Peng Y , Guo Y , Wang Q. Metabolic engineering of fast-growing Vibrio natriegens for efficient pyruvate production . Microb Cell Fact . 2023 ; 22 ( 1 ). doi: 10.1186/s12934-023-02185-0 OpenUrl CrossRef 12. ↵ Meng W , Zhang Y , Ma L , et al. Non-Sterilized Fermentation of 2,3-Butanediol with Seawater by Metabolic Engineered Fast-Growing Vibrio natriegens . Front Bioeng Biotechnol . 2022 ; 10 . doi: 10.3389/fbioe.2022.955097 OpenUrl CrossRef PubMed 13. ↵ Dalia TN , Hayes CA , Stolyar S , Marx CJ , McKinlay JB , Dalia AB . Multiplex Genome Editing by Natural Transformation (MuGENT) for Synthetic Biology in Vibrio natriegens . ACS Synth Biol . 2017 ; 6 ( 9 ). doi: 10.1021/acssynbio.7b00116 OpenUrl CrossRef PubMed 14. ↵ Ly Huong Tran . β-nicotinamide mononucleotide production in Vibrio natriegens: a preliminary study . Figshare . Published online 2025. doi: 10.6084/m9.figshare.28547603.v1 OpenUrl CrossRef 15. ↵ Weinstock MT , Hesek ED , Wilson CM , Gibson DG . Vibrio natriegens as a fast-growing host for molecular biology . Nat Methods . 2016 ; 13 ( 10 ). doi: 10.1038/nmeth.3970 OpenUrl CrossRef PubMed 16. ↵ Becker W , Wimberger F , Zangger K. Vibrio natriegens: An Alternative Expression System for the High-Yield Production of Isotopically Labeled Proteins . Biochemistry . 2019 ; 58 ( 25 ). doi: 10.1021/acs.biochem.9b00403 OpenUrl CrossRef 17. ↵ Vu Thi Nhat Le , Vu Thi Trang , Hasara Savindi Rupasinghe , Nguyen Quang Huy , Tran Thi Bich Thao . Simultaneous determination of nicotinamide mononucleotide, niacinamide, and nicotinic acid in health supplements using liquid chromatography . The Vietnam Journal of Food Control . Published online 2023. View the discussion thread. Back to top Previous Next Posted March 18, 2025. 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Share β-nicotinamide mononucleotide production in Vibrio natriegens : a preliminary study Ly Huong Tran , Vu Thao Phuong Tran , Huy Tuan Dat Pham , Giang Tra Nguyen , Tuoi Thi Nghiem , Anh Duc Pham , Nga Quynh Pham , Thuy Thi Le , Ngoc Lan Nguyen , Tran Nhat Minh Dang , Thinh Huy Tran , Van Khanh Tran , Hoa Quang Le bioRxiv 2025.03.17.643733; doi: https://doi.org/10.1101/2025.03.17.643733 Share This Article: Copy Citation Tools β-nicotinamide mononucleotide production in Vibrio natriegens : a preliminary study Ly Huong Tran , Vu Thao Phuong Tran , Huy Tuan Dat Pham , Giang Tra Nguyen , Tuoi Thi Nghiem , Anh Duc Pham , Nga Quynh Pham , Thuy Thi Le , Ngoc Lan Nguyen , Tran Nhat Minh Dang , Thinh Huy Tran , Van Khanh Tran , Hoa Quang Le bioRxiv 2025.03.17.643733; doi: https://doi.org/10.1101/2025.03.17.643733 Citation Manager Formats BibTeX Bookends EasyBib EndNote (tagged) EndNote 8 (xml) Medlars Mendeley Papers RefWorks Tagged Ref Manager RIS Zotero Tweet Widget Facebook Like Google Plus One Subject Area Bioengineering Subject Areas All Articles Animal Behavior and Cognition (7635) Biochemistry (17697) Bioengineering (13894) Bioinformatics (41951) Biophysics (21455) Cancer Biology (18592) Cell Biology (25507) Clinical Trials (138) Developmental Biology (13380) Ecology (19903) Epidemiology (2067) Evolutionary Biology (24321) Genetics (15610) Genomics (22509) Immunology (17737) Microbiology (40398) Molecular Biology (17182) Neuroscience (88618) Paleontology (667) Pathology (2833) Pharmacology and Toxicology (4825) Physiology (7641) Plant Biology (15158) Scientific Communication and Education (2046) Synthetic Biology (4296) Systems Biology (9825) Zoology (2271)