Biodegradation of PBAT and PHB/PBAT by Aspergillus and Purpureocillium isolates

Biodegradation of PBAT and PHB/PBAT by Aspergillus and Purpureocillium isolates | 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 Biodegradation of PBAT and PHB/PBAT by Aspergillus and Purpureocillium isolates Miguel Fernandes, António A. Vicente, Andreia F. Salvador This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6829190/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 In recent years blends and composites of biodegradable polymers have been developed as alternatives to petroleum-based plastics. Polybutylene adipate terephthalate (PBAT) and polyhydroxybutyrate (PHB) are examples of polymers that received much attention due to their interesting mechanical properties. However, although PHB presents a high biodegradation potential, PBAT is quite difficult to biodegrade under environmental conditions, and consequently only few microorganisms are known with this capability. In this work, two fungi recently isolated from soil were characterized regarding their ability to biodegrade PBAT and PHB/PBAT films. The polymers were the sole carbon and energy source, and biodegradation assays were performed in monoculture and in coculture. Biodegradation was assessed in solid and liquid media. Clear zones formation was monitored during incubation in agar plates containing the polymers, and films weight loss were measured in incubations in liquid medium. The consumption of oxygen was also monitored to confirm biodegradation.These fungi could efficiently biodegrade PHB/PBAT films at similar extents. Remarkably, after 14 days of incubation, isolate 7 (assigned to Aspergillus pseudoflectus ) achieved 40.7 % (wt) biodegradation and isolate 9 (assigned to Purpureocillium lilacinum ) 43.4 % (wt), whereas the coculture biodegraded 40.6 % (wt). PBAT biodegradation was far more challenging, and the fungi took 287 days to reach similar biodegradation percentages (41.2 % by isolate 9 and 30.2 % by isolate 7). This is the first study reporting the biodegradation PHB/PBAT films by aerobic mesophilic fungi. These fungi are promising candidates for the development of PBAT biodegradation technologies, and bioremediation strategies. Polyhydroxybutyrate Polybutylene adipate terephthalate Aspergillus Purpureocillium coculture Introduction PBAT is produced from adipic acid, 1,4-butanediol and terephthalic acid, and has analogous mechanical properties to conventional low-density polyethylene (LDPE), which turn it a very interesting alternative (Yamamoto et al., 2005). It can be degraded at thermophilic temperatures under composting conditions (Kijchavengkul et al., 2010). However, in natural environments such as soil, the degradation is considerably slower or inexistent (Han et al., 2021). The slower biodegradation rates when compared for example with polyhydroxyalkanoates polymers, is related to the limited number of microorganisms capable of degrading PBAT in the environment, and to the difficult action of the enzymes on terephthalic acid ester bonds (Soulenthone et al., 2020). The degradation of PBAT is well studied with compost isolates, namely with Thermobifida fusca at high temperatures (55 °C), which can achieve almost complete degradation in 22 days (Witt et al., 2001). The Thermobifida fusca DSM 43793 was found to act as an initial degrader, converting PBAT into lower molecular mass products, which were further degraded by other microorganisms, as Thermobifida fusca is unable to metabolize those degradation products (Kleeberg et al., 2005; Witt et al., 2001). Under mesophilic temperatures only very few isolated aerobic microorganisms, such as Bacillus subtilis ATCC 21332, Bacillus pumilus NKCM 3201, Cryptococcus sp. MTCC 5455, Isaria sp. NKCM 1712 and Leptothrix sp. TB-71 demonstrated the capacity to biodegrade PBAT, but generally biodegradation rates are low and biodegradation is only partial (Aarthy et al., 2018; Kasuya et al., 2009; Muroi et al., 2017; Nakajima-Kambe et al., 2009a; Trinh Tan et al., 2008). The degradation is even slower in anaerobic environments and known microorganisms are rare. Hungatella hathewayi DSM 13479 and Pelosinus fermentans are examples of anaerobes that can biodegrade PBAT, although more than 14 days are necessary to biodegrade about 5 % (Biundo et al., 2016; Perz et al., 2016a). Several enzymes with PBAT degrading activity are known, acting on for example esters bonds of short acyl chains, or cleaving preferably adipic acid with 1,4 butanediol ester bonds than 1,4 butanediol with terephthalic acid ester bonds or even being capable to completely hydrolyse PBAT (Perz et al., 2016; Suzuki et al., 2014; Zumstein et al., 2017). Trinh Tan et al. (2008) indicated an exo-mechanism for enzymes expressed by Bacillus subtilis since they degraded especially the lower molecular weight units. However, for polybutylene adipate, Penicillium pinophilum presented an endo mechanism with no accumulation of minor oligomeric compounds during the degradation of the polymer chains (Trinh Tan et al., 2008), showing that depending on the microorganisms and their genomic capabilities, the mechanism of PBAT biodegradation can be quite different. Recently in our lab, we isolated two fungi from a soil microcosm apparently with PBAT degrading activity (Fernandes et al., 2024). In this work, both microbial isolates were tested and characterized regarding their PBAT and PHB/PBAT biodegrading profiles. Biodegradation tests were performed in mono and coculture, in liquid media and under mesophilic conditions. The enzymes potentially involved in PBAT biodegradation were investigated as well by using an in silico analysis. MATERIALS AND METHODS 1.2.1 Microorganisms: identification, storage and maintenance The two microorganisms used in this study were isolated from soil containing PHB/PBAT films as described by Fernandes et al. (2024). They both grow under aerobic and mesophilic conditions (30 °C). Sequencing results indicated that the isolates are fungi assigned to Aspergillus pseudoflectus (Isolate 7) and Purpureocillium lilacinum (Isolate 9), being closely related to Aspergillus pseudoflectus strain isolate CCMG111 (99.83 % similarity at the Internal transcribed spacer (ITS) level) and Purpureocillium lilacinum clone SF_357 (100 % similarity at the ITS level) (Fernandes et al., 2024). Before the assays, spores stock solutions were prepared as described by Simões et al. (2015). Briefly, spores from isolate 7 and isolate 9 were collected from ten-day-old pure cultures grown in Potato Dextrose Agar (VWR Chemicals) at 30 °C, by flooding the surface of the agar plates with 2 mL of TWS solution (0.85 % NaCl plus 0.05 % Tween 80) and shaking gently. The suspension was then homogenized by vortexing and used for large-scale production of spores. The final spore suspension was homogenized by vortexing before quantification using a Neubauer count chamber. Aliquots of spore suspension were cryopreserved at -80 °C in 20 % glycerol, to allow using the same spore suspension in all assays. The spores were pre-germinated, to stimulate their metabolic state. The germinated spores were prepared by inoculating the aliquots in nutrient broth (VWR Chemicals) for 18 h (time necessary for the germination of > 95 % of the spores). Before use, these suspensions were always washed twice to eliminate the carbon sources, by centrifugation for 30 s (3000 g), and resuspended in phosphate-buffered saline solution. This final suspension was homogenized by vortexing before quantification using a Neubauer count chamber, to control the concentration of spores used in the assays (2 x 10 4 mL -1 ). The ITS sequences obtained for isolates 7 and 9 were deposited in the European Nucleotide Archive (https://www.ebi.ac.uk/ena/browser/home) under the accession number PRJEB62456 with the identifiers OY740347 and OY740348, respectively. Isolate 7 and 9 were deposited in the culture collection MUM (http://www.micoteca.deb.uminho.pt/) with the identifiers MUM 24.49 and MUM 24.50, respectively. 1.2.2 Polymers films The plastic was a PHB/PBAT 55/45 % (wt/wt %) bilayer used in the form of film. The PHB was an experimental PHB grade (Biomer P309) supplied by Biomer (Krailling, Germany). The PBAT was a commercial aliphatic-aromatic copolyester-based polymer (EcoflexVR F blend C1200, a film-blowing grade). Neat PBAT films were produced by extrusion using the same material. Details about the co-extrusion of the bilayer film and the extrusion of the PBAT film can be found in the work from Teixeira et al., (2020). 1.2.3 Clear zone methodology A mineral salt medium supplemented with PBAT as the sole carbon source was prepared accordingly to Fernandes et al. (2024). Briefly, a solid mineral medium with agar and with micro and nanoparticles of PBAT was used to assess clear zones formation, which indicates the PBAT degradation. The two isolates were tested in this medium by spread-plate for the formation of clear zones, at different mesophilic temperatures, more precisely at 20 °C, 25 ºC, 30 ºC and 37 °C. 1.2.4 PHB/PBAT biodegradation assays in liquid medium – weight loss evaluation Biodegradation assays were performed using Erlenmeyer shake flasks (125 mL) capped with foam plugs. The shake flasks with 10 mL of medium were sterilized with saturated steam at 121 °C (1 bar) in an autoclave for 15 min. The medium composition consisted in a mineral medium described elsewhere (Fernandes et al., 2024). Each flask contained a PHB/PBAT film (2 x 2 cm) as the sole carbon and energy source, which was previously sterilized by soaking in ethanol, followed by rinsing it with sterile water, dried and weighted. A control assay without microorganisms was prepared (abiotic assay). After inoculation, the shake flasks were incubated at 28 °C with agitation (250 rpm, in a rotary shaker) for 14 days. After the incubation period, the non-degraded remaining films were recovered, rinsed with distilled water, and dried to a constant weight. The weight loss of the film was determined by subtracting its weight after incubation from its initial weight. Spores of the two isolates were inoculated (2 x 10 4 mL -1 ), and for the coculture assays equal concentration of each isolate was inoculated. An abiotic control assay was performed in which no microorganism was inoculated. An additional set of experiments was conducted to evaluate the biodegradation of PBAT film (2 x 2 cm) only, without PHB. These experiments were set-up as described above except that PHB was not added and PBAT was the only carbon source. Abiotic assays were prepared as described above. 1.2.5 PBAT biodegradation assays in liquid medium – monitoring oxygen consumption Measurements of oxygen (O 2 ) consumption were carried out using a closed system method with air as the initial atmosphere. Briefly, the experiments were carried out in serum bottles of 120 mL, sealed with aluminium caps, with air in the headspace. The assays were carried out in a working volume of 60 mL, containing a carbon-free medium, the one described in Fernandes et al., (2024) and PBAT films (1 cm x 4 cm). The medium was sterilized by autoclaving, the assays were prepared in triplicate, under the following conditions: 1) inoculation with isolate 7; 2) inoculation with isolate 9; 3) inoculation with the coculture (isolate 7 and isolate 9 together), 4) abiotic control (with PBAT but without microorganisms). In each bottle, 33 mg of fungi (plate cultivated) were added. Positive control assays containing the fungi but nutrient broth (VWR Chemicals) instead of PBAT were performed, to confirm the microbial activity of the isolates (in duplicate assays). Incubations were performed at 27 °C under agitation (200 rpm). The concentrations of O 2 in the microcosms were measured by drawing gas samples with a 500 µL gas tight syringe suitable for gas chromatography. The O 2 content was analysed through a gas chromatograph (Bruker Scion 456, Markham, ON, Canada), equipped with a thermal conductivity detector (TCD) at 130 °C, and a Molsieve column and Argon (30 mL/min) as the carrier gas (Ballesteros et al., 2022). Air was used as a standard for O 2 concentration calibration. 1.2.6 Search for enzymes potentially involved in PBAT biodegradation To identify the enzymes that might be involved in PBAT biodegradation in isolates 7 and 9, we first performed a literature search to collect information about the enzymes already described as involved in the degradation of PBAT by other microorganisms (Table 1). Those enzymes were used as reference to search for protein homologs in the genomes of species closed related to the isolates (once the genomes of isolates 7 and 9 are not sequenced). Table 1 Enzymes associated to PBAT biodegradation and respective taxonomic assignment, based in literature search. Enzyme Microorganism UniProt EC number References Polyesterase Pseudomonas pseudoalcaligenes W6R2Y2 EC 3.1.1.74 (Wallace et al., 2017) Hydrolase Rhodococcus fascians NKCM251 A0A7I8E2Z4 (Soulenthone et al., 2021) Cutinase Thermobifida cellulosilytica E9LVH8 EC 3.1.1.74 (Perz et al., 2016a) E9LVH9 Cutinase Thermobifida alba D4Q9N1 EC 3.1.1.1.74 (Thumarat et al., 2015) Cutinase Thermobifida alba F7IX06 EC 3.1.1.1.74 (Thumarat et al., 2012) Hydrolase Pelosinus fermentans DSM 17108 A0A0A0YMQ9 (Biundo et al., 2016) Esterase Clostridium botulinum UPI0006BCBA3E (Perz et al., 2016b) UPI0006BC7D7E Hydrolase Bacillus pumilus A0A1E1FNX8 (Muroi et al., 2017) Polyesterase Saccharomonospora viridis W0TJ64 (Kawai et al., 2014) Carboxylesterase uncultured bacterium A0A1C9T884 (Müller et al., 2017) Carboxylesterase uncultured bacterium A0A1C9T772 Carboxylesterase uncultured bacterium A0A1C9T7D1 Carboxylesterase uncultured bacterium A0A1C9T784 Carboxylesterase uncultured bacterium A0A1C9T7D3 Carboxylesterase uncultured bacterium A0A1C9T7G6 Cutinase Humicola insolens A0A075B5G4 EC 3.1.1.74 (Perz et al., 2016a) Cutinase Fusarium solani P00590 Novozym® 51032 (Zumstein et al., 2017) Esterase Hungatella hathewayi DSM 13479 UPI000731EA1F (Perz et al., 2016c) Hydrolase Thermobifida fusca DSM 43793 Q6A0I3 (Kleeberg et al., 2005; Müller et al., 2005) Q6A0I4 Cutinase Paraphoma sp. B47-9 A0A060N399 (Suzuki et al., 2014) Cutinase Saitozyma flava (Cryptococcus flavus) A0A0P0ZE81 (Watanabe et al., 2015) The search was made against the genomes of Purpureocillium lilacinum species (strains PLFJ-1, 36-1 and PLBJ-1, taxon ID 33203), and the Aspergillus genus (including 553 species, taxon ID 5052), which are available in public databases, because the genomes of the isolates used in this study are not sequenced. The in silico search of PBAT-degrading enzymes in the selected genomes was performed with M-PARTY, a bioinformatics tool developed in-house and available at Bioconda (https://bioconda.github.io/recipes/m-party/README.html). Briefly, this tool builds Hidden Markov Models (HMMs) based on reference proteins (Table 1), extends the reference protein datasets by including other protein sequences belonging to the same protein family (via information obtained automatically by accessing the KEGG database), and finds homologous proteins in FASTA protein datasets. In this case, the FASTA protein datasets were download from the UniProt database (Tax ID 33203 and 5052). The extension of the reference database is made for increasing the number of reference enzymes, and therefore the probability of finding similar enzymes in the proteomes of the microorganisms. 1.2.7 Statistical analysis All the biodegradation trials performed in liquid medium were analysed using the OriginPRO 2019b statistical program by one-way analysis of variance (ANOVA) followed by the Fisher LSD test, and statistical significance was accepted as p < 0.05 (95 % significance). RESULTS 1.3.1 Evaluation of PBAT biodegradation by clear zones formation The results obtained with the clear zone assays showed that both isolates are capable of biodegrading PBAT at 20, 25 and 30 °C, although at 30 °C clear zones were formed faster. At 37 °C no clear zones were formed as indicated in Table 2, showing that these microorganisms do not biodegrade PBAT at this temperature. The performance of both isolates regarding clear zones formation was similar (Table 2). Soulenthone et al. (2020) indicated similar outcomes with Rhodococus fascians strain NBRC 100625 and strain NKCM 2511, which formed clear zones in PBAT agar plates at temperatures ranging from 25 °C to 30 °C, but not at temperatures above 37 °C. Table 2 Effect of temperature on the clear zone formation in PBAT emulsified agar plates by the isolates after 7 days of incubation. -:no clearing zone formation, +: radius of clear zone smaller than 1 cm, ++: radius of clear zone between 1 and 3 cm. +++: radius of clear zone superior to 3 cm. All experiments were performed in triplicate 33 1.3.2 Polymers degradation in liquid media The two isolates could biodegrade PHB/PBAT films, although isolate 9 was slightly more efficient, degrading more than 43 %, while isolate 7 degraded about 41 % (Table 3). The average biodegradation rate for isolate 7 was 2.9 ± 0.38 %/day while for isolate 9 it was 3.1 ± 0.35 %/day. The coculture formed by the two isolates together was slightly less efficient (2.9 ± 0.04 %/day on average) than isolate 9 alone and was as efficient as isolate 7. Some weight loss could be measured in the abiotic assay, showing residual plastic degradation (Table 3). Differences between the biotic assays were not significantly different but were significantly different from the abiotic assays. Table 3 Degradation of PHB/PBAT films and PBAT films after 14 days of incubation at 30 °C in liquid medium. All experiments were performed in triplicate. 4 3.4 Microorganism (Isolate/taxonomic assignment) Average weight loss (%) PHB/PBAT film PBAT film Isolate 7 (Aspergillus sp .) 40.7 ± 5.3 a 2.0 ± 0.2 3 Isolate 9 (Purpureocillium sp.) 43.4 ± 4.3 a 2.4 ± 0.4 2 Coculture (isolates 7 and 9) 40.6 ± 0.6 a 2.6 ± 0.3 2 Control assay (without microorganisms) 3.8 ± 0.5 b 0.4 ± 0.3 1 Different letters or numbers correspond to statistically significant differences ( p < 0.05). The results show the lower biodegradation of PBAT comparing to PHB/PBAT at a temperature of 30 °C (Table 3). All the conditions were significantly different in relation to the control assay. PBAT biodegradation by isolate 7 was also significantly different from isolate 9 and the co-culture. The results of the incubations containing only PBAT films suggest that the weight loss in the first set of trials with the PHB/PBAT films (Table 3) was mainly due to the action of the fungi on the PHB layer because the weight loss with PBAT films was substantially lower. Remarkably, both isolates can biodegrade the two polymers, but when both PHB and PBAT are available, preference is given to PHB as the carbon source. 1.3.3 Polymers degradation in liquid media assessed by oxygen consumption Experiments in liquid media containing PBAT showed that most of the oxygen was consumed during the first 60 days in all conditions tested (Table 4). From thereafter, oxygen consumption continued but at a slower rate. Isolate 9 was more efficient, reaching 41.0 % PBAT biodegradation while the isolated 7 reached 31.2 %. The co-culture achieved 28.8 % biodegradation, and even in the control (without microorganisms), some oxygen consumption was detected (13.8 %), suggesting that residual carbon sources were inoculated together with the microorganisms. Isolate 7, was less efficient in PBAT biodegradation when compared to isolate 9 (particularly in liquid minimum media), which may explain the lowest mineralization obtained by the coculture. Nevertheless, photographic images show the colonization of PBAT by isolate 7, which is a required step preceding biodegradation (Figure S1). All the conditions were significantly different in relation to the control after 287 days and isolate 9 achieved the highest biodegradation that was also significantly different from the other conditions. The weight loss in the control assay without microorganisms (Table 3) cannot be attributed to biodegradation, therefore, we performed an additional experiment to investigate the effect of the sterilization on the weight of the films. We verified that the films are altered by autoclaving conditions which was reflected in weight loss, i.e., PHB films showing a 1.4 ± 0.3 % weight loss and with PBAT films 0.3 ± 0.2 % weight loss after sterilization by autoclaving. Table 4 Biodegradation of PBAT films by isolate 7, isolate 9 and the coculture, in liquid media, accessed by oxygen consumption, after 35 or 287 days. All experiments were performed in triplicate 6 35 Microorganism (Isolate/Genus) PBAT (mg) After 35 days After 287 days O 2 Consumed (mmol.L -1 ) O 2 Consumed (mmol) Mineralization (%) Mineralization without control (% O 2 Consumed (mmol.L -1 ) O 2 Consumed (mmol) Mineralization (%) Mineralization without control (%)* Isolate 7 (Aspergillus) 16 2.7 0.2 17.1 5.3 4.9 0.3 30.2 b 17.4 Isolate 9 ( Purpureocillium ) 16 4.2 0.3 27.0 15.2 6.1 0.4 41.2 c 28.4 Coculture 16 2.6 0.2 16.6 4.8 4.3 0.3 27.4 b 14.6 Control (no microorganisms) 16 1.9 0.1 11.8 2.2 0.1 12.8 a - Isolate 7 (Positive Control) - 3.1 0.2 - 2.1 0.1 - - Isolate 9 (Positive Control) - 4.7 0.3 - 3.4 0.2 - - Letters a, b and c correspond to statistically significant differences ( p < 0.05) *Calculated by subtracting the oxygen consumption in the control assay without microorganisms 1.3.4 Identification of enzymes potentially involved in PBAT biodegradation By using the M-PARTY bioinformatics tool, a total of 420 HMMs were obtained. Twelve different proteins in the genome of Purpureocillium lilacinum match those previously known to be involved in PBAT biodegradation. These proteins were found by 50 different HMMs, meaning that some proteins were detected more than once (Table 5). Although a positive match was obtained for 12 proteins, manual curation of the results revealed that the function of some of those was not the same as the enzymes present in the initial dataset (Table 1), corresponding to false positives. For instance, 6 of the positive hits were annotated as amino acid transporter transmembrane domain-containing protein, ubiquinone biosynthesis protein COQ4 mitochondrial (Coenzyme Q biosynthesis protein), and 4 as beta-lactamases (data not shown). Therefore, only the remaining 6 proteins were considered as a valid result and were identified as cutinases (4 enzymes) and acetylxylan esterases (2 enzymes). These results make sense since these two types of enzymes were present in the reference dataset, as it can be seen in Table 1. Because isolate 9 is very closely related to Purpureocillium lilacinum it is highly probable that similar enzymes could have been expressed by isolate 9 during the biodegradation of PBAT. Table 5 Enzymes from Purpureocillium lilacinum (tax ID 33203) identified as potential PBAT degrading enzymes 2 36 Purpureocillium lilacinum Uniprot ID Protein names Gene Names Microorganism Number of HMMs that identify the protein A0A179GYZ5 Cutinase VFPBJ_05098 Purpureocillium lilacinum 7 A0A179HT03 Cutinase VFPFJ_04300 6 A0A2U3DZK7 Cutinase PCL_02616 6 A0A179GM63 Cutinase PCL_04231 VFPBJ_06338 VFPFJ_11216 40 A0A2U3DU00 Acetylxylan esterase PCL_06909 1 A0A179H4M9 Acetylxylan esterase VFPFJ_07534 1 Regarding the search on microorganisms assigned to the Aspergillus genus, 84 HMMs could identify 669 proteins, and similarly to what was obtained for Purpureocillium lilacinum , after manual curation only 440 were considered as positive results. These included 8 acyltransferases, 405 cutinases, 13 esterases and 14 transesterase. The genes coding to these enzymes were found in the genomes of 79 different microorganisms belonging to the Aspergillus genus. The UniProt IDs of the enzymes are indicated in Table S1. Isolate 7 possibly expressed equivalent enzymes during PBAT biodegradation. This study describes PBAT biodegrading activities in microorganisms assigned to Purpureocillium lilacinum and Aspergillus pseudoflectus . The fact that several genes were found on the genomes of microorganisms belonging to those genera suggests their generalized potential to biodegrade PBAT. DISCUSSION This study describes two microorganisms isolated from soil with the ability to biodegrade PBAT films. These are fungi assigned to Purpureocillium lilacinum and Aspergillus pseudoflectus . There are not many microbes isolated from soil with this capability recognized (Table 1). Therefore, this discovery contributes for increasing the number of PBAT degraders and the knowledge on PBAT biodegradation by soil microorganisms, opening new avenues for the development of biotechnological solutions for PBAT biodegradation. The isolates biodegraded PHB/PBAT and PBAT films either in pure culture or in coculture (Tables 2,3,4), but the biodegradability efficiency varied much according to the cultivation conditions. For instance, in agar plates, temperature clearly influenced the clear zone formation (Table 2), indicating that both fungi are mesophilic degraders with ideal growth between 25 °C and 30 °C. In all incubations, and in liquid medium, it was possible to see a fast weight loss of the PHB/PBAT films (Table 3), however, with films containing only PBAT, the weight loss was inferior. The fact that the two isolates can biodegrade PBAT under this temperature range offers an advantage for the implementation of treatment strategies using these microbes. Note that, PBAT is known to be biodegradable at higher temperatures (> 58 ºC), and particularly under composting conditions, but environmental temperatures are usually lower, and therefore mesophilic treatment solutions are less expensive. By searching the genomes of closely related microbes, we could find several enzyme candidates that might act on PBAT. Several enzyme matches were found in the genome of closely related microorganisms (6 for Purpureocillium lilacinum and 440 to Aspergillus ). Nevertheless, we cannot be completely sure that our two isolates effectively contain those enzymes, or if in alternative they contain others that can even be more efficient than the enzymes already identified from PBAT degraders. To identify the PBAT degrading enzymes of the isolates identified here, responsible for the biodegradation, further work needs to be conducted, such as gene expression assays during the biodegradation of the PBAT films. To enlighten the mechanism of action, in future works the enzymes should be isolated and tested to verify which ester bonds are cleaved, and what is the efficiency and the substrate specificity. It has been indicated in other studies that the hydrolytic action of fungi and bacteria on PBAT may be repressed in the presence of easily accessible carbon sources such as glucose and fructose (Kasuya et al., 2009; Kleeberg et al., 1998: (Soulenthone et al., 2020). This type of phenomenon may explain the results and the PHB preference found here, being PHB a more easily biodegradable substrate. Also, the expression or activity of the PBAT enzymes could have been inhibited. It is possible that the fungi studied in this work are capable of degrading one or more specific bonds of the PBAT polymer and growing using one or more products as substrate. However, is also possible that these fungi cannot metabolize PBAT completely. For example, Bacillus subtilis ATCC 21332 was capable of easily degrading three small esters, namely tributyrin, dibutyl adipate and dibutyl terephthalate and also the aliphatic polyester, polybutylene adipate (Trinh Tan et al., 2008). Nevertheless, it was not capable of degrading the aromatic group polybutylene terephthalate. Other PBAT-degrading microorganism, namely Leptothrix sp. TB-71 used some adipic acid but not 1,4-butanediol (Nakajima-Kambe et al., 2009). The bacteria Bacillus pumilus NKCM 3201 and Thermobifida fusca DSM 43793 did not use any PBAT constituents as subtracts to grow (Kleeberg et al., 2005; Muroi et al., 2017). In contrast, a Rhodococcus bacteriumwas capable of degrading PBAT and its constituents although at different rates, 55 % for 1,4-butanediol, 5 % for adipic acid, 7 % for terephthalic acid, and 7 % for PBAT (Soulenthone et al., 2020). PBAT is composed of an aromatic and an aliphatic group. The existence of the aromatic group in the polyester chain increases the resistance due to the lower hydrolytic degradation rate when compared with polyester with just aliphatic groups such as polylactic acid and polyglycolic acid (Kijchavengkul et al., 2010). This particular group, decreases the chain flexibility, creating fewer susceptible bonds, and originating a steric interference effect to the access of the more vulnerable ester bonds (Van Krevelen and Te Nijenhuis, 2009). For example, a lipase from Pseudomonas sp. had more difficulty cleaving esters near terephthalic acid in PBAT structure when compared to esters containing only aliphatic sequences (Marten et al., 2005) . Thedecrease in oxygen concentration in abiotic assays (12.8 % in 287 days – Table 4) may be attributed to several different factors. Namely, oxygen dissolution in the liquid media, which is promoted over time due to the synergistic effect of temperature and agitation during the incubations; possible thermo-oxidative reactions, since PBAT is susceptible to thermal degradation, although it is -consider stable at low temperatures (Al-Itry et al., 2012; Signori et al., 2009); hydrolytic degradation, because PBAT contains ester bonds (-COO) that are vulnerable to attack and subsequent cleavage by water molecules (Muthuraj et al., 2015) and the resulting products may react with oxygen. Note that the sterilization process can also contribute to alter the oxygen concentrations in the liquid and headspace of the incubation bottles and facilitate the hydrolysis by water and probably other reactions such as thermo-oxidative degradation. Comparing with reports in the literature the efficiency of the two isolates in PBAT biodegradation is quite high (Table 3). For instance, microbes such as Bacillus subtilis and Leptothrix sp. TB-71 took the same or more time (21 or 14 days respectively) to biodegrade less PBAT (1.4 and 16 mg) (Nakajima-Kambe et al., 2009b; Trinh Tan et al., 2008). Indeed, fungi seem to offer advantages over bacteria regarding PBAT biodegradation. For instance, Muroi et al., (2016) found that fungi, especially Ascomycota have a role in PBAT biodegradation in soil under mesophilic conditions. Other studies indicate that bacteria ( Rhodococuss fascians ) at mesophilic temperatures presented a five times lower growth rate than fungi, suggesting that the primary degrading species of PBAT are fungi ( Isaria fumosorosea ) (Kasuya et al., 2009; Soulenthone et al., 2020). This evidence highlights the potential use of our two fungi isolates in PBAT degradation-related solutions since they are likely to be more successful than bacteria. CONCLUSION This is the first work to report the degradation of PBAT and PHB/PBAT films by novel isolates assigned to Purpureocillium lilacinum and Aspergillus pseudoflectus . The results indicate that the two microorganisms are aerobically mesophilic fungi capable of degrading solid PBAT, although the degradation was superior with PHB/PBAT films, indicating a clear preference for the PHB polymer. From a biotechnological perspective, these results highlight the need to optimize the growth conditions to maximize plastic’s biodegradation by these isolates and use them in plastic´s clean-up strategies. The understanding of the mechanism of degradation may also promote the industrial development of biodegradable plastics appropriate for enzymatic recycling, and the tailoring of enzymes with increased efficiency. From an environmental viewpoint, these microorganisms are optimal candidates to be used in bioremediation of soil contaminated with plastics, given their abilities to grow in soil and to biodegrade plastic’s films. Declarations Author Contribution M.F- Conduct all experiments and wrote the main manuscript text. A.F.S. and A.A.V. - did the supervision and reviewed the manuscript Acknowledgments This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/ 04469/2020 unit, and by LABBELS – Associate Laboratory in Biotechnology, Bioengineering and Microelectromechanical Systems, LA/P/ 0029/2020. Furthermore, this study was also supported by BioTecNorte operation (NORTE-01–0145- FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. The fellowship is supported by a Doctoral advanced training (call NORTE-69–2015–15) funded by the European Social Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. Finally, by the doctoral grant PD/BD/146195/2019 and the grant COVID/BD/153308/2023. I would like to give thanks to José Freitas, for his valuable help with the M-party bioinformatics tool. References Al-Itry, R., Lamnawar, K., and Maazouz, A. (2012). Improvement of thermal stability, rheological and mechanical properties of PLA, PBAT and their blends by reactive extrusion with functionalized epoxy. Polym. Degrad. Stab. 97 , 1898–1914. Ballesteros, L.F., Teixeira, J.A., and Cerqueira, M.A. (2022). Active Carboxymethyl Cellulose-Based Edible Coatings for the Extension of Fresh Goldenberries Shelf-Life. Horticulturae 8 . Fernandes, M., Salvador, A.F., and Vicente, A.A. (2024). Biodegradation of PHB/PBAT films and isolation of novel PBAT biodegraders from soil microbiomes. Chemosphere 362 , 142696. Kasuya, K. ichi, Ishii, N., Inoue, Y., Yazawa, K., Tagaya, T., Yotsumoto, T., Kazahaya, J. ichiro, and Nagai, D. (2009). Characterization of a mesophilic aliphatic-aromatic copolyester-degrading fungus. Polym. Degrad. Stab. 94 , 1190–1196. Kijchavengkul, T., Auras, R., Rubino, M., Selke, S., Ngouajio, M., and Fernandez, R.T. (2010). Biodegradation and hydrolysis rate of aliphatic aromatic polyester. Polym. Degrad. Stab. 95 , 2641–2647. Kleeberg, I., Hetz, C., Kroppenstedt, R.M., Müller, R.J., and Deckwer, W.D. (1998). Biodegradation of aliphatic-aromatic copolyesters by Thermomonospora fusca and other thermophilic compost isolates. Appl. Environ. Microbiol. 64 , 1731–1735. Kleeberg, I., Welzel, K., VandenHeuvel, J., Müller, R.-J., and Deckwer, W.-D. (2005). Characterization of a New Extracellular Hydrolase from Thermobifida fusca Degrading Aliphatic−Aromatic Copolyesters. Biomacromolecules 6 , 262–270. Van Krevelen, D.W., and Te Nijenhuis, K. (2009). Polymer Properties. D.W. Van Krevelen, and K.B.T.-P. of P. (Fourth E. Te Nijenhuis, eds. (Amsterdam: Elsevier), pp. 3–5. Marten, E., Müller, R.J., and Deckwer, W.D. (2005). Studies on the enzymatic hydrolysis of polyesters. II. Aliphatic-aromatic copolyesters. Polym. Degrad. Stab. 88 , 371–381. Muroi, F., Tachibana, Y., Kobayashi, Y., Sakurai, T., and Kasuya, K.I. (2016). Influences of poly(butylene adipate-co-terephthalate) on soil microbiota and plant growth. Polym. Degrad. Stab. 129 , 338–346. Muroi, F., Tachibana, Y., Soulenthone, P., Yamamoto, K., Mizuno, T., Sakurai, T., Kobayashi, Y., and Kasuya, K. (2017). Characterization of a poly(butylene adipate-co-terephthalate) hydrolase from the aerobic mesophilic bacterium Bacillus pumilus. Polym. Degrad. Stab. 137 , 11–22. Muthuraj, R., Misra, M., and Mohanty, A.K. (2015). Hydrolytic degradation of biodegradable polyesters under simulated environmental conditions. J. Appl. Polym. Sci. 132 . Nakajima-Kambe, T., Toyoshima, K., Saito, C., Takaguchi, H., Akutsu-Shigeno, Y., Sato, M., Miyama, K., Nomura, N., and Uchiyama, H. (2009). Rapid monomerization of poly(butylene succinate)-co-(butylene adipate) by Leptothrix sp. J. Biosci. Bioeng. 108 , 513–516. Perz, V., Bleymaier, K., Sinkel, C., Kueper, U., Bonnekessel, M., Ribitsch, D., and Guebitz, G.M. (2016). Substrate specificities of cutinases on aliphatic–aromatic polyesters and on their model substrates. N. Biotechnol. 33 , 295–304. Signori, F., Coltelli, M.B., and Bronco, S. (2009). Thermal degradation of poly(lactic acid) (PLA) and poly(butylene adipate-co-terephthalate) (PBAT) and their blends upon melt processing. Polym. Degrad. Stab. 94 , 74–82. Simões, L.C., Simões, M., and Lima, N. (2015). Kinetics of biofilm formation by drinking water isolated Penicillium expansum. Biofouling 31 , 349–362. Soulenthone, P., Tachibana, Y., Muroi, F., Suzuki, M., Ishii, N., Ohta, Y., and Kasuya, K. (2020). Characterization of a mesophilic actinobacteria that degrades poly(butylene adipate-co-terephthalate). Polym. Degrad. Stab. 181 , 109335. Suzuki, K., Noguchi, M.T., Shinozaki, Y., and Koitabashi, M. (2014). Purification , characterization , and cloning of the gene for a biodegradable plastic-degrading enzyme from Paraphoma -related fungal strain B47-9. 4457–4465. Teixeira, P.F., Covas, J.A., Suarez, M.J., Angulo, I., and Hilliou, L. (2020). Film Blowing of PHB-Based Systems for Home Compostable Food Packaging. Int. Polym. Process. 35 , 440–447. Trinh Tan, F., Cooper, D.G., Marić, M., and Nicell, J.A. (2008). Biodegradation of a synthetic co-polyester by aerobic mesophilic microorganisms. Polym. Degrad. Stab. 93 , 1479–1485. Zumstein, M.T., Rechsteiner, D., Roduner, N., Perz, V., Ribitsch, D., Guebitz, G.M., Kohler, H.-P.E., McNeill, K., and Sander, M. (2017). Enzymatic Hydrolysis of Polyester Thin Films at the Nanoscale: Effects of Polyester Structure and Enzyme Active-Site Accessibility. Environ. Sci. Technol. 51 , 7476–7485. Additional Declarations No competing interests reported. Supplementary Files SupplementarymaterialFINAL.docx 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. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-6829190","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":490153548,"identity":"903224db-ffed-4f43-bcc7-d0a4fa356456","order_by":0,"name":"Miguel Fernandes","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABBElEQVRIiWNgGAWjYJCCDxAqAcj6YQNkMDYeIKCDcQZMC+PMnjSQQAPxWph52A6DmXi16LYfftjwcc8deXP25MMfeHjO261tPwy0pcYmGpcWszNpho0znj0z3NnzLE1CwuJ28rYziUAtx9JyG3BpOZDD/pjnwGHGDTdyzBgMeG4nmx0AamFsOIxby/k3jM1/Dhy233Aj//OHBLZzyWbnHxLQciOHsZnhwOFEoC0MEgfYDtiZ3SBky41nho09Bw4nbzjzzEyysSc5wewG0JYEfH45n/yw4ceBw7Ybjic//vznh5292fn0hw8+1Njg1IIBEsEqE4hVDgL2pCgeBaNgFIyCkQEAD/9xbSGYdLYAAAAASUVORK5CYII=","orcid":"","institution":"Centre of Biological Engineering, University of Minho","correspondingAuthor":true,"prefix":"","firstName":"Miguel","middleName":"","lastName":"Fernandes","suffix":""},{"id":490153549,"identity":"dea13742-e991-439b-871a-c5ed20662494","order_by":1,"name":"António A. Vicente","email":"","orcid":"","institution":"Centre of Biological Engineering, University of Minho","correspondingAuthor":false,"prefix":"","firstName":"António","middleName":"A.","lastName":"Vicente","suffix":""},{"id":490153550,"identity":"b0f83d7c-3034-4591-a968-345335f01cec","order_by":2,"name":"Andreia F. Salvador","email":"","orcid":"","institution":"Centre of Biological Engineering, University of Minho","correspondingAuthor":false,"prefix":"","firstName":"Andreia","middleName":"F.","lastName":"Salvador","suffix":""}],"badges":[],"createdAt":"2025-06-05 12:38:13","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6829190/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6829190/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":87997560,"identity":"ef613c63-61ed-46d5-af09-ff5d4f352962","added_by":"auto","created_at":"2025-07-31 09:39:19","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":813967,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6829190/v1/68c227b3-4e12-4169-9561-bf31789b1799.pdf"},{"id":87580506,"identity":"750a1296-7cfc-46e6-8e1b-79703f56819f","added_by":"auto","created_at":"2025-07-25 12:42:35","extension":"docx","order_by":0,"title":"","display":"","copyAsset":false,"role":"supplement","size":2280721,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementarymaterialFINAL.docx","url":"https://assets-eu.researchsquare.com/files/rs-6829190/v1/9864986a30cda7275059aa69.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Biodegradation of PBAT and PHB/PBAT by Aspergillus and Purpureocillium isolates","fulltext":[{"header":"Introduction","content":"\u003cp\u003ePBAT is produced from adipic acid, 1,4-butanediol and terephthalic acid, and has analogous mechanical properties to conventional low-density polyethylene (LDPE), which turn it a very interesting alternative (Yamamoto et al., 2005). It can be degraded at thermophilic temperatures under composting conditions (Kijchavengkul et al., 2010). However, in natural environments such as soil, the degradation is considerably slower or inexistent (Han et al., 2021). The slower biodegradation rates when compared for example with polyhydroxyalkanoates polymers, is related to the limited number of microorganisms capable of degrading PBAT in the environment, and to the difficult action of the enzymes on terephthalic acid ester bonds (Soulenthone et al., 2020). The degradation of PBAT is well studied with compost isolates, namely with \u003cem\u003eThermobifida fusca\u003c/em\u003e at high temperatures (55 °C), which can achieve almost complete degradation in 22 days (Witt et al., 2001). The \u003cem\u003eThermobifida fusca\u003c/em\u003e DSM 43793 was found to act as an initial degrader, converting PBAT into lower molecular mass products, which were further degraded by other microorganisms, as \u003cem\u003eThermobifida fusca\u003c/em\u003e is unable to metabolize those degradation products (Kleeberg et al., 2005; Witt et al., 2001).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eUnder mesophilic temperatures only very few isolated aerobic microorganisms, such as \u003cem\u003eBacillus subtilis\u003c/em\u003e ATCC 21332, \u003cem\u003eBacillus pumilus\u003c/em\u003e NKCM 3201, \u003cem\u003eCryptococcus\u003c/em\u003e sp. MTCC 5455, \u003cem\u003eIsaria\u003c/em\u003e sp. NKCM 1712 and \u003cem\u003eLeptothrix\u003c/em\u003e sp. TB-71 demonstrated the capacity to biodegrade PBAT, but generally biodegradation rates are low and biodegradation is only partial (Aarthy et al., 2018; Kasuya et al., 2009; Muroi et al., 2017; Nakajima-Kambe et al., 2009a; Trinh Tan et al., 2008). The degradation is even slower in anaerobic environments and known microorganisms are rare. \u003cem\u003eHungatella hathewayi\u003c/em\u003e DSM 13479 and \u003cem\u003ePelosinus fermentans\u003c/em\u003e are examples of anaerobes that can biodegrade PBAT, although more than 14 days are necessary to biodegrade about 5 % (Biundo et al., 2016; Perz et al., 2016a).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eSeveral enzymes with PBAT degrading activity are known, acting on for example esters bonds of short acyl chains, or cleaving\u0026nbsp;preferably adipic acid with 1,4 butanediol ester bonds than 1,4 butanediol with terephthalic acid ester bonds or even being capable to\u0026nbsp;completely hydrolyse PBAT\u0026nbsp;(Perz et al., 2016; Suzuki et al., 2014; Zumstein et al., 2017).\u0026nbsp;Trinh Tan et al. (2008)\u0026nbsp;indicated an exo-mechanism for enzymes expressed by\u003cem\u003e\u0026nbsp;Bacillus subtilis\u003c/em\u003e since they degraded especially the lower molecular weight units. However, for polybutylene adipate, \u003cem\u003ePenicillium pinophilum\u0026nbsp;\u003c/em\u003epresented an endo mechanism with no accumulation of minor oligomeric compounds during the degradation of the polymer chains (Trinh Tan et al., 2008), showing that depending on the microorganisms and their genomic capabilities, the mechanism of PBAT biodegradation can be quite different.\u003c/p\u003e\n\u003cp\u003eRecently in our lab, we isolated two fungi from a soil microcosm apparently with PBAT degrading activity (Fernandes et al., 2024). In this work, both microbial isolates were tested and characterized regarding their PBAT and PHB/PBAT biodegrading profiles. Biodegradation tests were performed in mono and coculture, in liquid media and under mesophilic conditions. The enzymes potentially involved in PBAT biodegradation were investigated as well by using an \u003cem\u003ein silico\u0026nbsp;\u003c/em\u003eanalysis.\u0026nbsp;\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cp\u003e1.2.1 \u003cspan id=\"_Toc149052374\"\u003e\u0026nbsp;Microorganisms: identification, storage and maintenance\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003eThe two microorganisms used in this study were isolated from soil containing\u0026nbsp;PHB/PBAT\u0026nbsp;films as described by\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eFernandes et al. (2024). They both grow under aerobic and mesophilic conditions (30 \u0026deg;C). Sequencing results indicated that the isolates are fungi assigned to \u003cem\u003eAspergillus pseudoflectus\u0026nbsp;\u003c/em\u003e(Isolate 7) and \u003cem\u003ePurpureocillium lilacinum\u0026nbsp;\u003c/em\u003e(Isolate 9), being closely related to \u003cem\u003eAspergillus pseudoflectus\u0026nbsp;\u003c/em\u003estrain\u0026nbsp;isolate\u003cem\u003e\u0026nbsp;\u003c/em\u003eCCMG111\u003cem\u003e\u0026nbsp;\u003c/em\u003e(99.83 % similarity at the Internal transcribed spacer (ITS) level) and \u003cem\u003ePurpureocillium lilacinum\u0026nbsp;\u003c/em\u003eclone SF_357 (100 % similarity at the \u0026nbsp;ITS level) (Fernandes et al., 2024).\u003c/p\u003e\n\u003cp\u003eBefore the assays, spores stock solutions were prepared as described by Sim\u0026otilde;es et al. (2015). Briefly,\u0026nbsp;spores from isolate 7 and isolate 9\u003cem\u003e\u0026nbsp;\u003c/em\u003ewere collected from ten-day-old pure cultures grown in Potato Dextrose Agar\u0026nbsp;(VWR Chemicals)\u0026nbsp;at 30\u0026nbsp;\u0026deg;C, by flooding the surface of the agar plates with 2 mL of TWS solution (0.85 % NaCl plus 0.05 % Tween 80) and shaking gently. The suspension was then homogenized by vortexing and used for large-scale production of spores. The final spore suspension was homogenized by vortexing before quantification using a Neubauer count chamber. Aliquots of spore\u0026nbsp;suspension were cryopreserved\u0026nbsp;at\u0026nbsp;-80\u0026nbsp;\u0026deg;C in 20 % glycerol, to allow using the same spore\u0026nbsp;suspension in all assays. The spores were pre-germinated, to stimulate their metabolic state. The germinated spores were prepared by inoculating the aliquots in nutrient broth (VWR Chemicals) for 18 h (time necessary for the germination of \u0026gt; 95 % of the spores). Before use, these suspensions were always washed twice to eliminate the carbon sources, by centrifugation for 30 s (3000 g), and resuspended in\u0026nbsp;phosphate-buffered saline\u0026nbsp;solution.\u0026nbsp;This final suspension was homogenized by vortexing before quantification using a Neubauer count chamber, to control the concentration of spores used in the assays (2 x 10\u003csup\u003e4\u003c/sup\u003e mL\u003csup\u003e-1\u003c/sup\u003e). The ITS sequences obtained for isolates 7 and 9 were deposited in the European Nucleotide Archive (https://www.ebi.ac.uk/ena/browser/home) under the accession number PRJEB62456 with the identifiers OY740347 and OY740348, respectively. Isolate 7 and 9 were deposited in the culture collection MUM (http://www.micoteca.deb.uminho.pt/) with the identifiers MUM 24.49 and MUM 24.50, respectively.\u003c/p\u003e\n\u003cp\u003e1.2.2 \u003cspan id=\"_Toc149052375\"\u003e\u0026nbsp;Polymers films\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003eThe plastic was a PHB/PBAT 55/45 % (wt/wt %) bilayer used in the form of film. The PHB was an experimental PHB grade (Biomer P309) supplied by Biomer (Krailling, Germany). The PBAT was a commercial aliphatic-aromatic copolyester-based polymer (EcoflexVR F blend C1200, a film-blowing grade). Neat PBAT films were produced by extrusion using the same material. Details about the co-extrusion of the bilayer film and the extrusion of the PBAT film can be found in the work from Teixeira et al., (2020).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e1.2.3 \u003cspan id=\"_Toc149052376\"\u003e\u0026nbsp;Clear zone methodology\u003c/span\u003e\u003c/p\u003e\n\u003cp id=\"_Toc149052377\"\u003eA mineral salt medium supplemented with PBAT as the sole carbon source was prepared accordingly to Fernandes et al. (2024). Briefly, a solid mineral medium with agar and with micro and nanoparticles of PBAT was used to assess clear zones formation, which indicates the PBAT degradation. The two isolates were tested in this medium by spread-plate for the formation of clear zones, at different mesophilic temperatures, more precisely at 20 \u0026deg;C, 25 \u0026ordm;C, 30 \u0026ordm;C and 37 \u0026deg;C.\u003c/p\u003e\n\u003cp\u003e1.2.4 \u003cspan id=\"_Toc149052378\"\u003e\u0026nbsp;PHB/PBAT biodegradation assays in liquid medium \u0026ndash; weight loss evaluation\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003eBiodegradation assays were performed using Erlenmeyer shake flasks (125 mL) capped with foam plugs. The shake flasks with 10 mL of medium were sterilized with saturated steam at 121 \u0026deg;C (1 bar) in an autoclave for 15 min. The medium composition consisted in a mineral medium described elsewhere (Fernandes et al., 2024). Each flask contained a PHB/PBAT film (2 x 2 cm) as the sole carbon and energy source, which was previously sterilized by soaking in ethanol, followed by rinsing it with sterile water, dried and weighted. A control assay without microorganisms was prepared (abiotic assay). After inoculation, the shake flasks were incubated at 28\u0026nbsp;\u0026deg;C with agitation (250 rpm, in a rotary shaker) for 14 days. After the incubation period, the non-degraded remaining films were recovered, rinsed with distilled water, and dried to a constant weight. The weight loss of the film was determined by subtracting its weight after incubation from its initial weight.\u0026nbsp;Spores of the two isolates were inoculated (2 x 10\u003csup\u003e4\u0026nbsp;\u003c/sup\u003emL\u003csup\u003e-1\u003c/sup\u003e), and for the coculture assays equal concentration of each isolate was inoculated.\u0026nbsp;An abiotic control assay was performed in which no microorganism was inoculated.\u0026nbsp;An additional set of experiments was conducted to evaluate the biodegradation of PBAT film\u0026nbsp;(2 x 2 cm) only, without PHB. These experiments were set-up as described above except that PHB was not added and PBAT was the only carbon source. Abiotic assays were prepared as described above.\u003c/p\u003e\n\u003cp\u003e1.2.5 \u003cspan id=\"_Toc149052379\"\u003e\u0026nbsp;PBAT\u0026nbsp;\u003c/span\u003ebiodegradation assays in liquid medium \u0026ndash; monitoring oxygen consumption\u003c/p\u003e\n\u003cp\u003eMeasurements of oxygen (O\u003csub\u003e2\u003c/sub\u003e) consumption were carried out using a closed system method with air as the initial atmosphere. Briefly, the experiments were carried out in serum bottles of 120 mL, sealed with aluminium caps, with air in the headspace. The assays were carried out in a working volume of 60 mL, containing a carbon-free medium, the one described in Fernandes et al., (2024) and PBAT films (1 cm x 4 cm). The medium was sterilized by autoclaving, the assays were prepared in triplicate, under the following conditions: 1) inoculation with isolate 7; 2) inoculation with isolate 9; 3) inoculation with the coculture (isolate 7 and isolate 9 together), 4) abiotic control (with PBAT but without microorganisms). In each bottle, 33 mg of fungi (plate cultivated) were added. Positive control assays containing the fungi but\u0026nbsp;nutrient broth (VWR Chemicals) instead of PBAT were performed, to confirm the microbial activity of the isolates (in duplicate assays).\u0026nbsp;Incubations were performed at 27 \u0026deg;C under agitation (200 rpm). The concentrations of O\u003csub\u003e2\u003c/sub\u003e in the microcosms were measured by drawing gas samples with a 500 \u0026micro;L gas tight syringe suitable for gas chromatography. The O\u003csub\u003e2\u003c/sub\u003e content was analysed through a gas chromatograph (Bruker Scion 456, Markham, ON, Canada), equipped with a thermal conductivity detector (TCD) at 130 \u0026deg;C, and a Molsieve column and Argon (30 mL/min) as the carrier gas (Ballesteros et al., 2022). Air was used as a standard for O\u003csub\u003e2\u003c/sub\u003e concentration calibration.\u003c/p\u003e\n\u003cp\u003e1.2.6 \u003cspan id=\"_Toc149052380\"\u003e\u0026nbsp;Search for enzymes potentially involved in PBAT biodegradation\u0026nbsp;\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003eTo identify the enzymes that might be involved in PBAT biodegradation in isolates 7 and 9, we first performed a literature search to collect information about the enzymes already described as involved in the degradation of PBAT by other microorganisms (Table 1). Those enzymes were used as reference to search for protein homologs in the genomes of species closed related to the isolates (once the genomes of isolates 7 and 9 are not sequenced).\u003c/p\u003e\n\u003cp id=\"_Toc149130796\"\u003e\u003cstrong\u003eTable 1\u003c/strong\u003e Enzymes associated to PBAT biodegradation and respective taxonomic assignment, based in literature search.\u003c/p\u003e\n\u003cdiv align=\"center\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eEnzyme\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eMicroorganism\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eUniProt\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eEC number\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eReferences\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"32\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd height=\"31\" style=\"width: 0px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003ePolyesterase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cem\u003ePseudomonas pseudoalcaligenes\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eW6R2Y2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eEC 3.1.1.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e(Wallace et al., 2017)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"49\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHydrolase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cem\u003eRhodococcus fascians\u0026nbsp;\u003c/em\u003eNKCM251\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A7I8E2Z4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e(Soulenthone et al., 2021)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"49\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003eCutinase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cem\u003eThermobifida cellulosilytica\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eE9LVH8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eEC 3.1.1.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e(Perz et al., 2016a)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"33\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eE9LVH9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd height=\"33\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCutinase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cem\u003eThermobifida alba\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eD4Q9N1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eEC 3.1.1.1.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e(Thumarat et al., 2015)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"33\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCutinase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cem\u003eThermobifida alba\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eF7IX06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eEC 3.1.1.1.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e(Thumarat et al., 2012)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"33\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHydrolase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cem\u003ePelosinus fermentans\u0026nbsp;\u003c/em\u003eDSM 17108\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A0A0YMQ9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e(Biundo et al., 2016)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"49\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003eEsterase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cem\u003eClostridium botulinum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eUPI0006BCBA3E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e(Perz et al., 2016b)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"33\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eUPI0006BC7D7E\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd height=\"33\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eHydrolase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cem\u003eBacillus pumilus\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A1E1FNX8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e(Muroi et al., 2017)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"33\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003ePolyesterase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cem\u003eSaccharomonospora viridis\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003eW0TJ64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e(Kawai et al., 2014)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"43\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd height=\"31\" style=\"width: 0px;\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCarboxylesterase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003euncultured bacterium\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A1C9T884\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd rowspan=\"6\"\u003e\n \u003cp\u003e(M\u0026uuml;ller et al., 2017)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"33\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCarboxylesterase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cem\u003euncultured bacterium\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A1C9T772\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd height=\"33\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCarboxylesterase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cem\u003euncultured bacterium\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A1C9T7D1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd height=\"33\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCarboxylesterase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cem\u003euncultured bacterium\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A1C9T784\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd height=\"33\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCarboxylesterase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cem\u003euncultured bacterium\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A1C9T7D3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd height=\"33\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCarboxylesterase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cem\u003euncultured bacterium\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A1C9T7G6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd height=\"33\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCutinase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cem\u003eHumicola insolens\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A075B5G4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eEC 3.1.1.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e(Perz et al., 2016a)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"33\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCutinase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cem\u003eFusarium solani\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eP00590\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eNovozym\u0026reg; 51032\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e(Zumstein et al., 2017)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"33\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eEsterase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cem\u003eHungatella hathewayi\u0026nbsp;\u003c/em\u003eDSM 13479\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eUPI000731EA1F\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e(Perz et al., 2016c)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"49\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003eHydrolase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cem\u003eThermobifida fusca\u0026nbsp;\u003c/em\u003eDSM 43793\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eQ6A0I3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e(Kleeberg et al., 2005; M\u0026uuml;ller et al., 2005)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"38\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eQ6A0I4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd height=\"31\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCutinase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cem\u003eParaphoma\u0026nbsp;\u003c/em\u003esp.\u003cem\u003e\u0026nbsp;\u003c/em\u003eB47-9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A060N399\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e(Suzuki et al., 2014)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"33\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCutinase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cem\u003eSaitozyma flava (Cryptococcus flavus)\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A0P0ZE81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e(Watanabe et al., 2015)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"49\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eThe search was made against the genomes of \u003cem\u003ePurpureocillium lilacinum\u003c/em\u003e species (strains PLFJ-1, 36-1 and PLBJ-1, taxon ID 33203), and the Aspergillus genus (including 553 species, taxon ID 5052), which are available in public databases, because the genomes of the isolates used in this study are not sequenced. The \u003cem\u003ein silico\u0026nbsp;\u003c/em\u003esearch of PBAT-degrading enzymes in the selected genomes was performed with M-PARTY, a bioinformatics tool developed in-house and available at Bioconda (https://bioconda.github.io/recipes/m-party/README.html). Briefly, this tool builds Hidden Markov Models (HMMs) based on reference proteins (Table 1), extends the reference protein datasets by including other protein sequences belonging to the same protein family (via information obtained automatically by accessing the KEGG database), and finds homologous proteins in FASTA protein datasets. In this case, the FASTA protein datasets were download from the UniProt database (Tax ID 33203 and 5052). The extension of the reference database is made for increasing the number of reference enzymes, and therefore the probability of finding similar enzymes in the proteomes of the microorganisms.\u0026nbsp;\u003c/p\u003e\n\u003cp id=\"_Toc149052381\"\u003e1.2.7 \u0026nbsp;Statistical analysis\u003c/p\u003e\n\u003cp\u003eAll the biodegradation trials performed in liquid medium were analysed using the OriginPRO 2019b statistical program by one-way analysis of variance (ANOVA) followed by the Fisher LSD test, and statistical significance was accepted as\u0026nbsp;\u003cem\u003ep\u003c/em\u003e \u0026lt; 0.05 (95 % significance).\u003c/p\u003e"},{"header":"RESULTS","content":"\u003cp\u003e\u003cstrong\u003e1.3.1\u0026nbsp;\u003c/strong\u003e\u003cspan id=\"_Toc149052383\"\u003e\u003cstrong\u003e\u0026nbsp;Evaluation of PBAT biodegradation by clear zones formation\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003eThe results obtained with the clear zone assays showed that both isolates are capable of biodegrading PBAT at 20, 25 and 30 \u0026deg;C, although at 30 \u0026deg;C clear zones were formed faster. At 37 \u0026deg;C no clear zones were formed as indicated in Table 2, showing that these microorganisms do not biodegrade PBAT at this temperature. The performance of both isolates regarding clear zones formation was similar (Table 2). Soulenthone et al. (2020) indicated similar outcomes with \u003cem\u003eRhodococus\u0026nbsp;\u003c/em\u003e\u003cem\u003efascians\u0026nbsp;\u003c/em\u003estrain NBRC 100625 and strain NKCM 2511, which formed clear zones in PBAT agar plates at temperatures ranging from 25 \u0026deg;C to 30 \u0026deg;C, but not at temperatures above 37 \u0026deg;C.\u003c/p\u003e\n\u003cp id=\"_Toc149130797\"\u003e\u003cstrong\u003eTable 2\u0026nbsp;\u003c/strong\u003eEffect of temperature on the clear zone formation in PBAT emulsified agar plates by the isolates after 7 days of incubation. -:no clearing zone formation, +: radius of clear zone smaller than 1 cm, ++: radius of clear zone between 1 and 3 cm. +++: radius of clear zone superior to 3 cm. All experiments were performed in triplicate\u003cstrong\u003e33\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cimg 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\"\u003e\u003c/p\u003e\n\u003cp\u003e1.3.2 \u003cspan id=\"_Toc149052384\"\u003e\u0026nbsp;Polymers degradation in liquid media\u003c/span\u003e\u003c/p\u003e\n\u003cp\u003eThe two isolates could biodegrade PHB/PBAT films, although isolate 9 was slightly more efficient, degrading more than 43 %, while isolate 7 degraded about 41 % (Table 3). The average biodegradation rate for isolate 7 was 2.9 \u0026plusmn; 0.38 %/day while for isolate 9 it was 3.1 \u0026plusmn; 0.35 %/day. The coculture formed by the two isolates together was slightly less efficient (2.9 \u0026plusmn; 0.04 %/day on average) than isolate 9 alone and was as efficient as isolate 7. Some weight loss could be measured in the abiotic assay, showing residual plastic degradation (Table 3). Differences between the biotic assays were not significantly different but were significantly different from the abiotic assays.\u003c/p\u003e\n\u003cp id=\"_Toc149130798\"\u003e\u003cstrong\u003eTable 3\u0026nbsp;\u003c/strong\u003eDegradation of PHB/PBAT films and PBAT films after 14 days of incubation at 30 \u0026deg;C in liquid medium. All experiments were performed in triplicate. \u003cstrong\u003e4\u003c/strong\u003e\u003cstrong\u003e3.4\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv align=\"Left\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eMicroorganism (Isolate/taxonomic assignment)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eAverage weight loss (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003ePHB/PBAT film\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003ePBAT film\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eIsolate\u003cem\u003e\u0026nbsp;7 (Aspergillus\u0026nbsp;\u003c/em\u003esp\u003cem\u003e.)\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e40.7 \u0026plusmn; 5.3\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2.0 \u0026plusmn; 0.2\u003csup\u003e\u0026nbsp;\u003c/sup\u003e\u003csup\u003e3\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eIsolate\u003cem\u003e\u0026nbsp;9 (Purpureocillium sp.)\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e43.4 \u0026plusmn; 4.3\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2.4 \u0026plusmn; 0.4\u003csup\u003e\u0026nbsp;2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCoculture (isolates 7 and 9)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e40.6 \u0026plusmn; 0.6\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2.6 \u0026plusmn; 0.3\u003csup\u003e\u0026nbsp;\u003c/sup\u003e\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eControl assay (without microorganisms)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3.8 \u0026plusmn; 0.5\u003csup\u003e\u0026nbsp;\u003c/sup\u003e\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.4 \u0026plusmn; 0.3\u003csup\u003e1\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eDifferent letters or numbers correspond to statistically significant differences (\u003cem\u003ep\u003c/em\u003e \u0026lt; 0.05).\u003c/p\u003e\n\u003cp\u003eThe results show the lower biodegradation of PBAT comparing to PHB/PBAT at a temperature of 30 \u0026deg;C (Table 3). All the conditions were significantly different in relation to the control assay. PBAT biodegradation by isolate 7\u003cem\u003e\u0026nbsp;\u003c/em\u003ewas also significantly different from isolate 9 and the co-culture. The results of the incubations containing only PBAT films suggest that the weight loss in the first set of trials with the PHB/PBAT films (Table 3) was mainly due to the action of the fungi on the PHB layer because the weight loss with PBAT films was substantially lower. Remarkably, both isolates can biodegrade the two polymers, but when both PHB and PBAT are available, preference is given to PHB as the carbon source.\u003c/p\u003e\n\u003cp\u003e1.3.3 \u0026nbsp;Polymers degradation in liquid media assessed by oxygen consumption\u003c/p\u003e\n\u003cp\u003eExperiments in liquid media containing PBAT showed that most of the oxygen was consumed during the first 60 days in all conditions tested (Table 4). From thereafter, oxygen consumption continued but at a slower rate. Isolate 9 was more efficient, reaching 41.0 % PBAT biodegradation while the isolated 7 reached 31.2 %. The co-culture achieved 28.8 % biodegradation, and even in the control (without microorganisms), some oxygen consumption was detected (13.8 %), suggesting that residual carbon sources were inoculated together with the microorganisms. Isolate 7, was less efficient in PBAT biodegradation when compared to isolate 9 (particularly in liquid minimum media), which may explain the lowest mineralization obtained by the coculture. Nevertheless, photographic images show the colonization of PBAT by isolate 7, which is a required step preceding biodegradation (Figure S1). \u0026nbsp;All the conditions were significantly different in relation to the control after 287 days and isolate 9 achieved the highest biodegradation that was also significantly different from the other conditions. The weight loss in the control assay without microorganisms (Table 3) cannot be attributed to biodegradation, therefore, we performed an additional experiment to investigate the effect of the sterilization on the weight of the films. We verified that the films are altered by autoclaving conditions which was reflected in weight loss, i.e., PHB films showing a 1.4 \u0026plusmn; 0.3 % weight loss and with PBAT films 0.3 \u0026plusmn; 0.2 % weight loss after sterilization by autoclaving.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 4\u0026nbsp;\u003c/strong\u003eBiodegradation of PBAT films by isolate 7, isolate 9 and the coculture, in liquid media, accessed by oxygen consumption, after 35 or 287 days. All experiments were performed in triplicate \u003cstrong\u003e6\u003c/strong\u003e\u003cstrong\u003e35\u003c/strong\u003e\u003c/p\u003e\n\u003cdiv align=\"center\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eMicroorganism (Isolate/Genus)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003ePBAT (mg)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\"\u003e\n \u003cp\u003e\u003cstrong\u003eAfter 35 days\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\"\u003e\n \u003cp\u003e\u003cstrong\u003eAfter 287 days\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eO\u003csub\u003e2\u003c/sub\u003e Consumed (mmol.L\u003csup\u003e-1\u003c/sup\u003e)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eO\u003csub\u003e2\u003c/sub\u003e Consumed (mmol)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eMineralization (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eMineralization without control (%\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eO\u003csub\u003e2\u003c/sub\u003e Consumed (mmol.L\u003csup\u003e-1\u003c/sup\u003e)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eO\u003csub\u003e2\u003c/sub\u003e Consumed (mmol)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eMineralization (%)\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e\u003cstrong\u003eMineralization without control (%)*\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eIsolate\u003cem\u003e\u0026nbsp;\u003c/em\u003e7\u003cem\u003e\u0026nbsp;(Aspergillus)\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e17.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e5.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e30.2\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e17.4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eIsolate 9 (\u003cem\u003ePurpureocillium\u003c/em\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e27.0\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e15.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e6.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e41.2\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e28.4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eCoculture\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e16.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e27.4\u003csup\u003e\u0026nbsp;\u003c/sup\u003e\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e14.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eControl (no microorganisms)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e11.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e12.8\u003csup\u003e\u0026nbsp;a\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eIsolate\u0026nbsp;7 (Positive\u0026nbsp;Control)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e2.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eIsolate 9 (Positive Control)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e4.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e3.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"10\"\u003e\n \u003cp\u003eLetters a, b and c correspond to statistically significant differences (\u003cem\u003ep\u003c/em\u003e \u0026lt; 0.05)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026nbsp;*Calculated by subtracting the oxygen consumption in the control assay without microorganisms\u003c/p\u003e\n\u003cp id=\"_Toc149052386\"\u003e1.3.4 Identification of enzymes potentially involved in PBAT biodegradation\u003c/p\u003e\n\u003cp\u003eBy using the M-PARTY bioinformatics tool, a total of 420 HMMs were obtained. Twelve different proteins in the genome of \u003cem\u003ePurpureocillium lilacinum\u003c/em\u003e match those previously known to be involved in PBAT biodegradation. These proteins were found by 50 different HMMs, meaning that some proteins were detected more than once (Table 5). Although a positive match was obtained for 12 proteins, manual curation of the results revealed that the function of some of those was not the same as the enzymes present in the initial dataset (Table 1), corresponding to false positives. For instance, 6 of the positive hits were annotated as amino acid transporter transmembrane domain-containing protein, ubiquinone biosynthesis protein COQ4 mitochondrial (Coenzyme Q biosynthesis protein), and 4 as beta-lactamases (data not shown). Therefore, only the remaining 6 proteins were considered as a valid result and were identified as cutinases (4 enzymes) and acetylxylan esterases (2 enzymes). These results make sense since these two types of enzymes were present in the reference dataset, as it can be seen in Table 1. Because isolate 9 is very closely related to \u003cem\u003ePurpureocillium lilacinum\u0026nbsp;\u003c/em\u003eit is highly probable that similar enzymes could have been expressed by isolate 9 during the biodegradation of PBAT.\u003c/p\u003e\n\u003cp id=\"_Toc149130800\"\u003e\u003cstrong\u003eTable 5\u0026nbsp;\u003c/strong\u003eEnzymes from Purpureocillium lilacinum (tax ID 33203) identified as potential PBAT degrading enzymes \u003cstrong\u003e2\u003c/strong\u003e\u003cstrong\u003e36\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"5\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u003cem\u003ePurpureocillium lilacinum\u003cu\u003e\u0026nbsp;\u003c/u\u003e\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"21\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eUniprot ID\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eProtein names\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eGene Names\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eMicroorganism\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\"\u003e\n \u003cp\u003e\u003cstrong\u003eNumber of HMMs that identify the protein\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd height=\"39\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd height=\"21\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A179GYZ5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eCutinase\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eVFPBJ_05098\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"6\"\u003e\n \u003cp\u003e\u003cem\u003ePurpureocillium lilacinum\u003c/em\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd height=\"30\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A179HT03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eCutinase\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eVFPFJ_04300\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd height=\"30\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A2U3DZK7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eCutinase\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePCL_02616\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd height=\"30\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A179GM63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eCutinase\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePCL_04231 VFPBJ_06338 VFPFJ_11216\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd height=\"75\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A2U3DU00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eAcetylxylan esterase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003ePCL_06909\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd height=\"30\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd\u003e\n \u003cp\u003eA0A179H4M9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eAcetylxylan esterase\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003eVFPFJ_07534\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd height=\"31\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eRegarding the search on microorganisms assigned to the \u003cem\u003eAspergillus\u003c/em\u003e genus, 84 HMMs could identify 669 proteins, and similarly to what was obtained for \u003cem\u003ePurpureocillium lilacinum\u003c/em\u003e, after manual curation only 440 were considered as positive results. These included 8 acyltransferases, 405 cutinases, 13 esterases and 14 transesterase. The genes coding to these enzymes were found in the genomes of 79 different microorganisms belonging to the \u003cem\u003eAspergillus\u003c/em\u003e genus. The UniProt IDs of the enzymes are indicated in Table S1. Isolate 7 possibly expressed equivalent enzymes during PBAT biodegradation.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThis study describes PBAT biodegrading activities in microorganisms assigned to \u003cem\u003ePurpureocillium lilacinum\u003c/em\u003e and \u003cem\u003eAspergillus\u003c/em\u003e \u003cem\u003epseudoflectus\u003c/em\u003e. The fact that several genes were found on the genomes of microorganisms belonging to those genera suggests their generalized potential to biodegrade PBAT.\u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eThis study describes two microorganisms isolated from soil with the ability to biodegrade PBAT films. These are fungi assigned to\u0026nbsp;\u003cem\u003ePurpureocillium lilacinum\u003c/em\u003e and \u003cem\u003eAspergillus pseudoflectus\u003c/em\u003e\u003cem\u003e.\u0026nbsp;\u003c/em\u003eThere are not many microbes isolated from soil with this capability recognized (Table 1). Therefore, this discovery contributes for increasing the number of PBAT degraders and the knowledge on PBAT biodegradation by soil microorganisms, opening new avenues for the development of biotechnological solutions for PBAT biodegradation.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe isolates biodegraded PHB/PBAT and PBAT films either in pure culture or in coculture (Tables 2,3,4), but the biodegradability efficiency varied much according to the cultivation conditions. For instance, in agar plates, temperature clearly influenced the clear zone formation (Table 2), indicating that both fungi are mesophilic degraders with ideal growth between 25\u0026nbsp;°C and 30\u0026nbsp;°C. In all incubations,\u0026nbsp;and in liquid medium, it was possible to see a fast weight loss of the PHB/PBAT films (Table 3), however, with films containing only PBAT, the weight loss was inferior. The fact that the two isolates can biodegrade PBAT under this temperature range offers an advantage for the implementation of treatment strategies using these microbes. Note that, PBAT is known to be biodegradable at higher temperatures (\u0026gt; 58 ºC), and particularly under composting conditions, but environmental temperatures are usually lower, and therefore mesophilic treatment solutions are less expensive.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eBy searching the genomes of closely related microbes, we could find several enzyme candidates that might act on PBAT. Several enzyme matches were found in the genome of closely related microorganisms (6 for\u0026nbsp;\u003cem\u003ePurpureocillium lilacinum\u003c/em\u003e and 440 to\u0026nbsp;\u003cem\u003eAspergillus\u003c/em\u003e). Nevertheless, we cannot be completely sure that our two isolates effectively contain those enzymes, or if in alternative they contain others that can even be more efficient than the enzymes already identified from PBAT degraders.\u0026nbsp;To identify the PBAT degrading enzymes of the isolates identified here, responsible for the biodegradation, further work needs to be conducted, such as gene expression assays during the biodegradation of the PBAT films. To\u0026nbsp;enlighten the mechanism of action, in future works the enzymes should be isolated and tested to verify which ester bonds are cleaved, and what is the efficiency and the substrate specificity.\u003c/p\u003e\n\u003cp\u003eIt has been indicated in other studies that the hydrolytic action of fungi and bacteria on PBAT may be repressed in the presence of easily accessible carbon sources such as glucose and fructose (Kasuya et al., 2009; Kleeberg et al., 1998: (Soulenthone et al., 2020). This type of phenomenon may explain the results and the PHB preference found here, being PHB a more easily biodegradable substrate. Also, the expression or activity of the PBAT enzymes could have been inhibited. It is possible that the fungi studied in this work are capable of degrading one or more specific bonds of the PBAT polymer and growing using one or more products as substrate. However, is also possible that these fungi cannot metabolize PBAT completely. For example, \u003cem\u003eBacillus subtilis\u003c/em\u003e ATCC 21332 was capable of easily degrading three small esters, namely tributyrin, dibutyl adipate and dibutyl terephthalate and also the aliphatic polyester, polybutylene adipate (Trinh Tan et al., 2008). Nevertheless, it was not capable of degrading the aromatic group polybutylene terephthalate. Other PBAT-degrading microorganism, namely \u003cem\u003eLeptothrix\u003c/em\u003e sp. TB-71 used some adipic acid but not 1,4-butanediol (Nakajima-Kambe et al., 2009). The bacteria \u003cem\u003eBacillus pumilus\u003c/em\u003e NKCM 3201 and \u003cem\u003eThermobifida fusca\u0026nbsp;\u003c/em\u003eDSM 43793 did not use any PBAT constituents as subtracts to grow (Kleeberg et al., 2005; Muroi et al., 2017). In contrast, a \u003cem\u003eRhodococcus\u0026nbsp;\u003c/em\u003ebacteriumwas capable of degrading PBAT and its constituents although at different rates, 55 % for 1,4-butanediol, 5 % for\u0026nbsp;adipic acid, 7 % for terephthalic acid, and 7 % for PBAT\u0026nbsp;(Soulenthone et al., 2020).\u0026nbsp;PBAT is composed of an aromatic and an aliphatic group. The existence\u0026nbsp;of the\u0026nbsp;aromatic group in the polyester chain increases the resistance due to the lower hydrolytic degradation rate when compared with polyester with just aliphatic groups such as polylactic acid and polyglycolic acid\u0026nbsp;(Kijchavengkul et al., 2010). This particular group, decreases the chain flexibility, creating fewer susceptible bonds, and originating a steric interference effect to the access of the more vulnerable ester bonds\u0026nbsp;(Van Krevelen and Te Nijenhuis, 2009). For example, a lipase\u0026nbsp;from\u0026nbsp;\u003cem\u003ePseudomonas\u003c/em\u003e sp.\u0026nbsp;had more difficulty cleaving esters near terephthalic acid in PBAT structure when\u0026nbsp;compared to esters containing only aliphatic sequences\u0026nbsp;(Marten et al., 2005)\u003cstrong\u003e.\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThedecrease in oxygen concentration in abiotic assays (12.8 % in 287 days – Table 4) may be attributed to several different factors. Namely, oxygen dissolution in the liquid media, which is promoted over time due to the synergistic effect of temperature and agitation during the incubations; possible thermo-oxidative reactions, since PBAT is susceptible to thermal degradation, although it is -consider stable at low temperatures (Al-Itry et al., 2012; Signori et al., 2009); hydrolytic degradation, because PBAT contains ester bonds (-COO) that are vulnerable to attack and subsequent cleavage by water molecules (Muthuraj et al., 2015) and the resulting products may react with oxygen. Note that the sterilization process can also contribute to alter the oxygen concentrations in the liquid and headspace of the incubation bottles and facilitate the hydrolysis by water and probably other reactions such as thermo-oxidative degradation.\u003c/p\u003e\n\u003cp\u003eComparing with reports in the literature the efficiency of the two isolates in PBAT biodegradation is quite high (Table 3). For instance, microbes such as \u003cem\u003eBacillus subtilis\u0026nbsp;\u003c/em\u003eand \u003cem\u003eLeptothrix\u003c/em\u003e sp. TB-71 took the same or more time (21 or 14 days respectively) to biodegrade less PBAT (1.4 and 16 mg) (Nakajima-Kambe et al., 2009b; Trinh Tan et al., 2008).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIndeed, fungi seem to offer advantages over bacteria regarding PBAT biodegradation. For instance,\u0026nbsp;Muroi et al., (2016) found that fungi, especially \u003cem\u003eAscomycota\u003c/em\u003e have a role in PBAT biodegradation in soil under mesophilic conditions. Other studies\u0026nbsp;indicate that bacteria (\u003cem\u003eRhodococuss fascians\u003c/em\u003e) at mesophilic temperatures presented a five times lower growth rate than fungi, suggesting that the primary degrading species of PBAT are fungi (\u003cem\u003eIsaria fumosorosea\u003c/em\u003e) (Kasuya et al., 2009; Soulenthone et al., 2020). This evidence highlights the potential use of our two fungi isolates in PBAT degradation-related solutions since they are likely to be more successful than bacteria.\u0026nbsp;\u003c/p\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eThis is the first work to report the degradation of PBAT and PHB/PBAT films by novel isolates assigned to \u003cem\u003ePurpureocillium lilacinum\u003c/em\u003e and \u003cem\u003eAspergillus pseudoflectus\u003c/em\u003e. The results indicate that the two microorganisms are\u0026nbsp;aerobically mesophilic\u0026nbsp;fungi capable of\u0026nbsp;degrading solid PBAT,\u0026nbsp;although the degradation was superior with PHB/PBAT films, indicating a clear preference for the PHB polymer.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eFrom a biotechnological perspective, these results highlight the need to optimize the growth conditions to maximize plastic’s biodegradation by these isolates and use them in plastic´s clean-up strategies. The understanding of the mechanism of degradation may also promote the industrial development of biodegradable plastics appropriate for enzymatic recycling, and the tailoring of enzymes with increased efficiency. From an environmental viewpoint, these microorganisms are optimal candidates to be used in bioremediation of soil contaminated with plastics, given their abilities to grow in soil and to biodegrade plastic’s films.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eM.F- Conduct all experiments and wrote the main manuscript text. A.F.S. and A.A.V. - did the supervision and reviewed the manuscript\u003c/p\u003e\u003ch2\u003eAcknowledgments\u003c/h2\u003e\u003cp\u003eThis study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/ 04469/2020 unit, and by LABBELS \u0026ndash; Associate Laboratory in Biotechnology, Bioengineering and Microelectromechanical Systems, LA/P/ 0029/2020. Furthermore, this study was also supported by BioTecNorte operation (NORTE-01\u0026ndash;0145- FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. The fellowship is supported by a Doctoral advanced training (call NORTE-69\u0026ndash;2015\u0026ndash;15) funded by the European Social Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. Finally, by the doctoral grant PD/BD/146195/2019 and the grant COVID/BD/153308/2023. I would like to give thanks to Jos\u0026eacute; Freitas, for his valuable help with the M-party bioinformatics tool.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAl-Itry, R., Lamnawar, K., and Maazouz, A. (2012). Improvement of thermal stability, rheological and mechanical properties of PLA, PBAT and their blends by reactive extrusion with functionalized epoxy. Polym. Degrad. Stab. \u003cem\u003e97\u003c/em\u003e, 1898\u0026ndash;1914.\u003c/li\u003e\n \u003cli\u003eBallesteros, L.F., Teixeira, J.A., and Cerqueira, M.A. (2022). Active Carboxymethyl Cellulose-Based Edible Coatings for the Extension of Fresh Goldenberries Shelf-Life. Horticulturae \u003cem\u003e8\u003c/em\u003e.\u003c/li\u003e\n \u003cli\u003eFernandes, M., Salvador, A.F., and Vicente, A.A. (2024). Biodegradation of PHB/PBAT films and isolation of novel PBAT biodegraders from soil microbiomes. Chemosphere \u003cem\u003e362\u003c/em\u003e, 142696.\u003c/li\u003e\n \u003cli\u003eKasuya, K. ichi, Ishii, N., Inoue, Y., Yazawa, K., Tagaya, T., Yotsumoto, T., Kazahaya, J. ichiro, and Nagai, D. (2009). Characterization of a mesophilic aliphatic-aromatic copolyester-degrading fungus. Polym. Degrad. Stab. \u003cem\u003e94\u003c/em\u003e, 1190\u0026ndash;1196.\u003c/li\u003e\n \u003cli\u003eKijchavengkul, T., Auras, R., Rubino, M., Selke, S., Ngouajio, M., and Fernandez, R.T. (2010). Biodegradation and hydrolysis rate of aliphatic aromatic polyester. Polym. Degrad. Stab. \u003cem\u003e95\u003c/em\u003e, 2641\u0026ndash;2647.\u003c/li\u003e\n \u003cli\u003eKleeberg, I., Hetz, C., Kroppenstedt, R.M., M\u0026uuml;ller, R.J., and Deckwer, W.D. (1998). Biodegradation of aliphatic-aromatic copolyesters by Thermomonospora fusca and other thermophilic compost isolates. Appl. Environ. Microbiol. \u003cem\u003e64\u003c/em\u003e, 1731\u0026ndash;1735.\u003c/li\u003e\n \u003cli\u003eKleeberg, I., Welzel, K., VandenHeuvel, J., M\u0026uuml;ller, R.-J., and Deckwer, W.-D. (2005). Characterization of a New Extracellular Hydrolase from Thermobifida fusca Degrading Aliphatic\u0026minus;Aromatic Copolyesters. Biomacromolecules \u003cem\u003e6\u003c/em\u003e, 262\u0026ndash;270.\u003c/li\u003e\n \u003cli\u003eVan Krevelen, D.W., and Te Nijenhuis, K. (2009). Polymer Properties. D.W. Van Krevelen, and K.B.T.-P. of P. (Fourth E. Te Nijenhuis, eds. (Amsterdam: Elsevier), pp. 3\u0026ndash;5.\u003c/li\u003e\n \u003cli\u003eMarten, E., M\u0026uuml;ller, R.J., and Deckwer, W.D. (2005). Studies on the enzymatic hydrolysis of polyesters. II. Aliphatic-aromatic copolyesters. Polym. Degrad. Stab. \u003cem\u003e88\u003c/em\u003e, 371\u0026ndash;381.\u003c/li\u003e\n \u003cli\u003eMuroi, F., Tachibana, Y., Kobayashi, Y., Sakurai, T., and Kasuya, K.I. (2016). Influences of poly(butylene adipate-co-terephthalate) on soil microbiota and plant growth. Polym. Degrad. Stab. \u003cem\u003e129\u003c/em\u003e, 338\u0026ndash;346.\u003c/li\u003e\n \u003cli\u003eMuroi, F., Tachibana, Y., Soulenthone, P., Yamamoto, K., Mizuno, T., Sakurai, T., Kobayashi, Y., and Kasuya, K. (2017). Characterization of a poly(butylene adipate-co-terephthalate) hydrolase from the aerobic mesophilic bacterium Bacillus pumilus. Polym. Degrad. Stab. \u003cem\u003e137\u003c/em\u003e, 11\u0026ndash;22.\u003c/li\u003e\n \u003cli\u003eMuthuraj, R., Misra, M., and Mohanty, A.K. (2015). Hydrolytic degradation of biodegradable polyesters under simulated environmental conditions. J. Appl. Polym. Sci. \u003cem\u003e132\u003c/em\u003e.\u003c/li\u003e\n \u003cli\u003eNakajima-Kambe, T., Toyoshima, K., Saito, C., Takaguchi, H., Akutsu-Shigeno, Y., Sato, M., Miyama, K., Nomura, N., and Uchiyama, H. (2009). Rapid monomerization of poly(butylene succinate)-co-(butylene adipate) by Leptothrix sp. J. Biosci. Bioeng. \u003cem\u003e108\u003c/em\u003e, 513\u0026ndash;516.\u003c/li\u003e\n \u003cli\u003ePerz, V., Bleymaier, K., Sinkel, C., Kueper, U., Bonnekessel, M., Ribitsch, D., and Guebitz, G.M. (2016). Substrate specificities of cutinases on aliphatic\u0026ndash;aromatic polyesters and on their model substrates. N. Biotechnol. \u003cem\u003e33\u003c/em\u003e, 295\u0026ndash;304.\u003c/li\u003e\n \u003cli\u003eSignori, F., Coltelli, M.B., and Bronco, S. (2009). Thermal degradation of poly(lactic acid) (PLA) and poly(butylene adipate-co-terephthalate) (PBAT) and their blends upon melt processing. Polym. Degrad. Stab. \u003cem\u003e94\u003c/em\u003e, 74\u0026ndash;82.\u003c/li\u003e\n \u003cli\u003eSim\u0026otilde;es, L.C., Sim\u0026otilde;es, M., and Lima, N. (2015). Kinetics of biofilm formation by drinking water isolated Penicillium expansum. Biofouling \u003cem\u003e31\u003c/em\u003e, 349\u0026ndash;362.\u003c/li\u003e\n \u003cli\u003eSoulenthone, P., Tachibana, Y., Muroi, F., Suzuki, M., Ishii, N., Ohta, Y., and Kasuya, K. (2020). Characterization of a mesophilic actinobacteria that degrades poly(butylene adipate-co-terephthalate). Polym. Degrad. Stab. \u003cem\u003e181\u003c/em\u003e, 109335.\u003c/li\u003e\n \u003cli\u003eSuzuki, K., Noguchi, M.T., Shinozaki, Y., and Koitabashi, M. (2014). Purification , characterization , and cloning of the gene for a biodegradable plastic-degrading enzyme from Paraphoma -related fungal strain B47-9. 4457\u0026ndash;4465.\u003c/li\u003e\n \u003cli\u003eTeixeira, P.F., Covas, J.A., Suarez, M.J., Angulo, I., and Hilliou, L. (2020). Film Blowing of PHB-Based Systems for Home Compostable Food Packaging. Int. Polym. Process. \u003cem\u003e35\u003c/em\u003e, 440\u0026ndash;447.\u003c/li\u003e\n \u003cli\u003eTrinh Tan, F., Cooper, D.G., Marić, M., and Nicell, J.A. (2008). Biodegradation of a synthetic co-polyester by aerobic mesophilic microorganisms. Polym. Degrad. Stab. \u003cem\u003e93\u003c/em\u003e, 1479\u0026ndash;1485.\u003c/li\u003e\n \u003cli\u003eZumstein, M.T., Rechsteiner, D., Roduner, N., Perz, V., Ribitsch, D., Guebitz, G.M., Kohler, H.-P.E., McNeill, K., and Sander, M. (2017). Enzymatic Hydrolysis of Polyester Thin Films at the Nanoscale: Effects of Polyester Structure and Enzyme Active-Site Accessibility. Environ. Sci. Technol. \u003cem\u003e51\u003c/em\u003e, 7476\u0026ndash;7485.\u003c/li\u003e\n\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":"Polyhydroxybutyrate, Polybutylene adipate terephthalate, Aspergillus, Purpureocillium, coculture","lastPublishedDoi":"10.21203/rs.3.rs-6829190/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6829190/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIn recent years blends and composites of biodegradable polymers have been developed as alternatives to petroleum-based plastics. Polybutylene adipate terephthalate (PBAT) and polyhydroxybutyrate (PHB) are examples of polymers that received much attention due to their interesting mechanical properties. However, although PHB presents a high biodegradation potential, PBAT is quite difficult to biodegrade under environmental conditions, and consequently only few microorganisms are known with this capability. In this work, two fungi recently isolated from soil were characterized regarding their ability to biodegrade PBAT and PHB/PBAT films. The polymers were the sole carbon and energy source, and biodegradation assays were performed in monoculture and in coculture. Biodegradation was assessed in solid and liquid media. Clear zones formation was monitored during incubation in agar plates containing the polymers, and films weight loss were measured in incubations in liquid medium. The consumption of oxygen was also monitored to confirm biodegradation.These fungi could efficiently biodegrade PHB/PBAT films at similar extents. Remarkably, after 14 days of incubation, isolate 7 (assigned to \u003cem\u003eAspergillus pseudoflectus\u003c/em\u003e) achieved 40.7 % (wt) biodegradation and isolate 9 (assigned to \u003cem\u003ePurpureocillium lilacinum\u003c/em\u003e) 43.4 % (wt), whereas the coculture biodegraded 40.6 % (wt). PBAT biodegradation was far more challenging, and the fungi took 287 days to reach similar biodegradation percentages (41.2 % by isolate 9 and 30.2 % by isolate 7).\u003c/p\u003e\n\u003cp\u003eThis is the first study reporting the biodegradation PHB/PBAT films by aerobic mesophilic fungi. These fungi are promising candidates for the development of PBAT biodegradation technologies, and bioremediation strategies.\u003c/p\u003e","manuscriptTitle":"Biodegradation of PBAT and PHB/PBAT by Aspergillus and Purpureocillium isolates","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-25 12:34:30","doi":"10.21203/rs.3.rs-6829190/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":"9f01f91f-2147-4d46-8d27-a01f1c5ba8c7","owner":[],"postedDate":"July 25th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-07-31T09:39:06+00:00","versionOfRecord":[],"versionCreatedAt":"2025-07-25 12:34:30","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6829190","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6829190","identity":"rs-6829190","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}