Anna N Khusnutdinova, Hai Tran, Saloni Devlekar, Marco A Distaso, Ilya V Kublanov, Tatiana Skarina, Peter Stogios, Alexei Savchenko, Manuel Ferrer, Olga V Golyshina, Alexander F Yakunin, Peter N Golyshin
{"title":"来自超嗜酸性古菌 Cuniculiplasma divulgatum S5 的中等恒温 GH1 β-葡萄糖苷酶。","authors":"Anna N Khusnutdinova, Hai Tran, Saloni Devlekar, Marco A Distaso, Ilya V Kublanov, Tatiana Skarina, Peter Stogios, Alexei Savchenko, Manuel Ferrer, Olga V Golyshina, Alexander F Yakunin, Peter N Golyshin","doi":"10.1093/femsec/fiae114","DOIUrl":null,"url":null,"abstract":"<p><p>Family GH1 glycosyl hydrolases are ubiquitous in prokaryotes and eukaryotes and are utilized in numerous industrial applications, including bioconversion of lignocelluloses. In this study, hyperacidophilic archaeon Cuniculiplasma divulgatum (S5T=JCM 30642T) was explored as a source of novel carbohydrate-active enzymes. The genome of C. divulgatum encodes three GH1 enzyme candidates, from which CIB12 and CIB13 were heterologously expressed and characterized. Phylogenetic analysis of CIB12 and CIB13 clustered them with β-glucosidases from genuinely thermophilic archaea including Thermoplasma acidophilum, Picrophilus torridus, Sulfolobus solfataricus, Pyrococcus furiosus, and Thermococcus kodakarensis. Purified enzymes showed maximal activities at pH 4.5-6.0 (CIB12) and 4.5-5.5 (CIB13) with optimal temperatures at 50°C, suggesting a high-temperature origin of Cuniculiplasma spp. ancestors. Crystal structures of both enzymes revealed a classical (α/β)8 TIM-barrel fold with the active site located inside the barrel close to the C-termini of β-strands including the catalytic residues Glu204 and Glu388 (CIB12), and Glu204 and Glu385 (CIB13). Both enzymes preferred cellobiose over lactose as substrates and were classified as cellobiohydrolases. Cellobiose addition increased the biomass yield of Cuniculiplasma cultures growing on peptides by 50%, suggesting that the cellobiohydrolases expand the carbon substrate range and hence environmental fitness of Cuniculiplasma.</p>","PeriodicalId":12312,"journal":{"name":"FEMS microbiology ecology","volume":" ","pages":""},"PeriodicalIF":3.5000,"publicationDate":"2024-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11376072/pdf/","citationCount":"0","resultStr":"{\"title\":\"Moderately thermostable GH1 β-glucosidases from hyperacidophilic archaeon Cuniculiplasma divulgatum S5.\",\"authors\":\"Anna N Khusnutdinova, Hai Tran, Saloni Devlekar, Marco A Distaso, Ilya V Kublanov, Tatiana Skarina, Peter Stogios, Alexei Savchenko, Manuel Ferrer, Olga V Golyshina, Alexander F Yakunin, Peter N Golyshin\",\"doi\":\"10.1093/femsec/fiae114\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Family GH1 glycosyl hydrolases are ubiquitous in prokaryotes and eukaryotes and are utilized in numerous industrial applications, including bioconversion of lignocelluloses. In this study, hyperacidophilic archaeon Cuniculiplasma divulgatum (S5T=JCM 30642T) was explored as a source of novel carbohydrate-active enzymes. The genome of C. divulgatum encodes three GH1 enzyme candidates, from which CIB12 and CIB13 were heterologously expressed and characterized. Phylogenetic analysis of CIB12 and CIB13 clustered them with β-glucosidases from genuinely thermophilic archaea including Thermoplasma acidophilum, Picrophilus torridus, Sulfolobus solfataricus, Pyrococcus furiosus, and Thermococcus kodakarensis. Purified enzymes showed maximal activities at pH 4.5-6.0 (CIB12) and 4.5-5.5 (CIB13) with optimal temperatures at 50°C, suggesting a high-temperature origin of Cuniculiplasma spp. ancestors. Crystal structures of both enzymes revealed a classical (α/β)8 TIM-barrel fold with the active site located inside the barrel close to the C-termini of β-strands including the catalytic residues Glu204 and Glu388 (CIB12), and Glu204 and Glu385 (CIB13). 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Moderately thermostable GH1 β-glucosidases from hyperacidophilic archaeon Cuniculiplasma divulgatum S5.
Family GH1 glycosyl hydrolases are ubiquitous in prokaryotes and eukaryotes and are utilized in numerous industrial applications, including bioconversion of lignocelluloses. In this study, hyperacidophilic archaeon Cuniculiplasma divulgatum (S5T=JCM 30642T) was explored as a source of novel carbohydrate-active enzymes. The genome of C. divulgatum encodes three GH1 enzyme candidates, from which CIB12 and CIB13 were heterologously expressed and characterized. Phylogenetic analysis of CIB12 and CIB13 clustered them with β-glucosidases from genuinely thermophilic archaea including Thermoplasma acidophilum, Picrophilus torridus, Sulfolobus solfataricus, Pyrococcus furiosus, and Thermococcus kodakarensis. Purified enzymes showed maximal activities at pH 4.5-6.0 (CIB12) and 4.5-5.5 (CIB13) with optimal temperatures at 50°C, suggesting a high-temperature origin of Cuniculiplasma spp. ancestors. Crystal structures of both enzymes revealed a classical (α/β)8 TIM-barrel fold with the active site located inside the barrel close to the C-termini of β-strands including the catalytic residues Glu204 and Glu388 (CIB12), and Glu204 and Glu385 (CIB13). Both enzymes preferred cellobiose over lactose as substrates and were classified as cellobiohydrolases. Cellobiose addition increased the biomass yield of Cuniculiplasma cultures growing on peptides by 50%, suggesting that the cellobiohydrolases expand the carbon substrate range and hence environmental fitness of Cuniculiplasma.
期刊介绍:
FEMS Microbiology Ecology aims to ensure efficient publication of high-quality papers that are original and provide a significant contribution to the understanding of microbial ecology. The journal contains Research Articles and MiniReviews on fundamental aspects of the ecology of microorganisms in natural soil, aquatic and atmospheric habitats, including extreme environments, and in artificial or managed environments. Research papers on pure cultures and in the areas of plant pathology and medical, food or veterinary microbiology will be published where they provide valuable generic information on microbial ecology. Papers can deal with culturable and non-culturable forms of any type of microorganism: bacteria, archaea, filamentous fungi, yeasts, protozoa, cyanobacteria, algae or viruses. In addition, the journal will publish Perspectives, Current Opinion and Controversy Articles, Commentaries and Letters to the Editor on topical issues in microbial ecology.
- Application of ecological theory to microbial ecology
- Interactions and signalling between microorganisms and with plants and animals
- Interactions between microorganisms and their physicochemical enviornment
- Microbial aspects of biogeochemical cycles and processes
- Microbial community ecology
- Phylogenetic and functional diversity of microbial communities
- Evolutionary biology of microorganisms