دورية أكاديمية
Thermophilic Carboxylesterases from Hydrothermal Vents of the Volcanic Island of Ischia Active on Synthetic and Biobased Polymers and Mycotoxins.
| العنوان: | Thermophilic Carboxylesterases from Hydrothermal Vents of the Volcanic Island of Ischia Active on Synthetic and Biobased Polymers and Mycotoxins. |
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| المؤلفون: | Distaso MA; Centre for Environmental Biotechnology, School of Natural Sciences, Bangor University, Bangor, United Kingdom., Chernikova TN; Centre for Environmental Biotechnology, School of Natural Sciences, Bangor University, Bangor, United Kingdom., Bargiela R; Centre for Environmental Biotechnology, School of Natural Sciences, Bangor University, Bangor, United Kingdom., Coscolín C; Department of Applied Biocatalysis, ICP, CSIC, Madrid, Spain., Stogios P; Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Canada., Gonzalez-Alfonso JL; Department of Applied Biocatalysis, ICP, CSIC, Madrid, Spain., Lemak S; Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Canada., Khusnutdinova AN; Centre for Environmental Biotechnology, School of Natural Sciences, Bangor University, Bangor, United Kingdom., Plou FJ; Department of Applied Biocatalysis, ICP, CSIC, Madrid, Spain., Evdokimova E; Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Canada., Savchenko A; Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Canada.; Department of Microbiology Immunology and Infectious Diseases, University of Calgary, Calgary, Canada., Lunev EA; Centre for Environmental Biotechnology, School of Natural Sciences, Bangor University, Bangor, United Kingdom.; Institute of Gene Biology, Russian Academy of Sciences, Moscow, Russia., Yakimov MM; Institute of Polar Sciences, National Research Council, Messina, Italy., Golyshina OV; Centre for Environmental Biotechnology, School of Natural Sciences, Bangor University, Bangor, United Kingdom., Ferrer M; Department of Applied Biocatalysis, ICP, CSIC, Madrid, Spain., Yakunin AF; Centre for Environmental Biotechnology, School of Natural Sciences, Bangor University, Bangor, United Kingdom.; Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Canada., Golyshin PN; Centre for Environmental Biotechnology, School of Natural Sciences, Bangor University, Bangor, United Kingdom. |
| المصدر: | Applied and environmental microbiology [Appl Environ Microbiol] 2023 Feb 28; Vol. 89 (2), pp. e0170422. Date of Electronic Publication: 2023 Jan 31. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: American Society for Microbiology Country of Publication: United States NLM ID: 7605801 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1098-5336 (Electronic) Linking ISSN: 00992240 NLM ISO Abbreviation: Appl Environ Microbiol Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Washington, American Society for Microbiology. |
| مواضيع طبية MeSH: | Carboxylic Ester Hydrolases*/metabolism , Hydrothermal Vents*, Polymers ; Hydrolases/metabolism ; Polyesters ; Plastics ; Substrate Specificity |
| مستخلص: | Hydrothermal vents are geographically widespread and host microorganisms with robust enzymes useful in various industrial applications. We examined microbial communities and carboxylesterases of two terrestrial hydrothermal vents of the volcanic island of Ischia (Italy) predominantly composed of Firmicutes , Proteobacteria , and Bacteroidota . High-temperature enrichment cultures with the polyester plastics polyhydroxybutyrate and polylactic acid (PLA) resulted in an increase of Thermus and Geobacillus species and to some extent Fontimonas and Schleiferia species. The screening at 37 to 70°C of metagenomic fosmid libraries from above enrichment cultures identified three hydrolases (IS10, IS11, and IS12), all derived from yet-uncultured Chloroflexota and showing low sequence identity (33 to 56%) to characterized enzymes. Enzymes expressed in Escherichia coli exhibited maximal esterase activity at 70 to 90°C, with IS11 showing the highest thermostability (90% activity after 20-min incubation at 80°C). IS10 and IS12 were highly substrate promiscuous and hydrolyzed all 51 monoester substrates tested. Enzymes were active with PLA, polyethylene terephthalate model substrate, and mycotoxin T-2 (IS12). IS10 and IS12 had a classical α/β-hydrolase core domain with a serine hydrolase catalytic triad (Ser155, His280, and Asp250) in their hydrophobic active sites. The crystal structure of IS11 resolved at 2.92 Å revealed the presence of a N-terminal β-lactamase-like domain and C-terminal lipocalin domain. The catalytic cleft of IS11 included catalytic Ser68, Lys71, Tyr160, and Asn162, whereas the lipocalin domain enclosed the catalytic cleft like a lid and contributed to substrate binding. Our study identified novel thermotolerant carboxylesterases with a broad substrate range, including polyesters and mycotoxins, for potential applications in biotechnology. IMPORTANCE High-temperature-active microbial enzymes are important biocatalysts for many industrial applications, including recycling of synthetic and biobased polyesters increasingly used in textiles, fibers, coatings and adhesives. Here, we identified three novel thermotolerant carboxylesterases (IS10, IS11, and IS12) from high-temperature enrichment cultures from Ischia hydrothermal vents and incubated with biobased polymers. The identified metagenomic enzymes originated from uncultured Chloroflexota and showed low sequence similarity to known carboxylesterases. Active sites of IS10 and IS12 had the largest effective volumes among the characterized prokaryotic carboxylesterases and exhibited high substrate promiscuity, including hydrolysis of polyesters and mycotoxin T-2 (IS12). Though less promiscuous than IS10 and IS12, IS11 had a higher thermostability with a high temperature optimum (80 to 90°C) for activity and hydrolyzed polyesters, and its crystal structure revealed an unusual lipocalin domain likely involved in substrate binding. The polyesterase activity of these enzymes makes them attractive candidates for further optimization and potential application in plastics recycling. |
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| فهرسة مساهمة: | Keywords: 3PET; Ischia; PLA; biochemical characterization; carboxylesterase; crystal structure; hydrothermal vents; metagenome screening; polyesterase; thermophilic bacteria |
| المشرفين على المادة: | EC 3.1.1.- (Carboxylic Ester Hydrolases) 0 (Polymers) 0 (polyhydroxybutyrate) EC 3.- (Hydrolases) 0 (Polyesters) 0 (Plastics) |
| تواريخ الأحداث: | Date Created: 20230131 Date Completed: 20230302 Latest Revision: 20230313 |
| رمز التحديث: | 20230314 |
| مُعرف محوري في PubMed: | PMC9972953 |
| DOI: | 10.1128/aem.01704-22 |
| PMID: | 36719236 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1098-5336 |
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| DOI: | 10.1128/aem.01704-22 |