Moderately thermostable GH1 β-glucosidases from hyperacidophilic archaeon Cuniculiplasma divulgatum S5.

IF 3.5 3区 生物学 Q2 MICROBIOLOGY
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
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引用次数: 0

Abstract

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.

来自超嗜酸性古菌 Cuniculiplasma divulgatum S5 的中等恒温 GH1 β-葡萄糖苷酶。
GH1 糖基水解酶家族在原核生物和真核生物中无处不在,并被广泛应用于工业领域,包括木质纤维素的生物转化。本研究将超嗜酸古菌 Cuniculiplasma divulgatum(S5T=JCM 30642T)作为新型碳水化合物活性酶的来源进行了探索。C. divulgatum 的基因组编码了三种候选 GH1 酶,其中 CIB12 和 CIB13 已被异源表达和鉴定。CIB12 和 CIB13 的系统发育分析将它们与真正的嗜热古细菌(包括嗜酸热原体、嗜酸热微菌、溶血硫球菌、呋喃焦球菌和柯达卡伦热球菌)中的 β-葡萄糖苷酶进行了聚类。纯化的酶在 pH 值为 4.5-6.0 时(CIB12)和 4.5-5.5 时(CIB13)显示出最大活性,最佳温度为 50 °C,这表明 Cuniculiplasma 属祖先起源于高温。两种酶的晶体结构均显示出经典的(α/β)8 TIM桶状折叠,活性位点位于桶状结构内部,靠近包括催化残基Glu204和Glu388(CIB12)以及Glu204和Glu385(CIB13)在内的β-链的C端。这两种酶都喜欢以纤维生物糖而不是乳糖为底物,因此被归类为纤维生物水解酶。添加纤维素生物糖可使在肽上生长的 Cuniculiplasma 培养物的生物量增加 50%,这表明纤维素生物水解酶扩大了碳底物的范围,从而提高了 Cuniculiplasma 的环境适应性。
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来源期刊
FEMS microbiology ecology
FEMS microbiology ecology 生物-微生物学
CiteScore
7.50
自引率
2.40%
发文量
132
审稿时长
3 months
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
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