title: Improvement of the catalytic performance of a hyperthermostable GH10 xylanase from Talaromyces leycettanus JCM12802.
abstract: A xylanase gene of GH 10, Tlxyn10A, was cloned from Talaromyces leycettanus JCM12802 and expressed in Pichia pastoris. Purified recombinant TlXyn10A was acidic and hyperthermophilic, and retained stable over the pH range of 2.0-6.0 and at 90°C. Sequence analysis of TlXyn10A identified seven residues probably involved in substrate contacting. Three mutants (TlXyn10A_P, _N and _C) were then constructed by substituting some or all of the residues with corresponding ones of hyperthermal Xyl10C from Bispora sp. MEY-1. TlXyn10A_P with mutations at subsites +2 to +4 exhibited improved specific activity (by 0.44-fold) and pH stability (2.0-10.0). Molecular dynamics simulation analysis indicated that mutations E229I and F232E probably weaken the substrate affinity at subsites +3 to +4, and G149D may introduce a new hydrogen bond. These modifications altogether account for the improved performance of TlXyn10A_P. Moreover, TlXyn10A_P was able to hydrolyze wheat straw persistently, and has the application potentials in various industries.
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China; College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, People's Republic of China.
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Wang
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
authors.forename: Huoqing
authors.initials: H
authors.lastname: Huang
authors.affiliation: College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, People's Republic of China.
authors.forename: Xiangming
authors.initials: X
authors.lastname: Xie
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
authors.forename: Rui
authors.initials: R
authors.lastname: Ma
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
authors.forename: Yingguo
authors.initials: Y
authors.lastname: Bai
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China; College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, People's Republic of China.
authors.forename: Fei
authors.initials: F
authors.lastname: Zheng
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
authors.forename: Shuai
authors.initials: S
authors.lastname: You
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
authors.forename: Bingyu
authors.initials: B
authors.lastname: Zhang
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
authors.forename: Huifang
authors.initials: H
authors.lastname: Xie
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China.
authors.forename: Bin
authors.initials: B
authors.lastname: Yao
authors.affiliation: Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, People's Republic of China. Electronic address: luohuiying@caas.cn.
authors.forename: Huiying
authors.initials: H
authors.lastname: Luo
mesh_terms: D000595:Amino Acid Sequence; D055162:Biocatalysis / Q000187:drug effects; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D043364:Endo-1,4-beta Xylanases / Q000378:metabolism*; D004795:Enzyme Stability / Q000187:drug effects; D006031:Glycosylation; D006868:Hydrolysis; D007477:Ions; D007700:Kinetics; D008670:Metals / Q000494:pharmacology; D056004:Molecular Dynamics Simulation; D009154:Mutation / Q000235:genetics; D011994:Recombinant Proteins / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D040681:Structural Homology, Protein; D013379:Substrate Specificity / Q000187:drug effects; D032901:Talaromyces / Q000201:enzymology* / Q000235:genetics; D013696:Temperature; D014908:Triticum / Q000737:chemistry; D014866:Waste Products; D014990:Xylans / Q000378:metabolism
chemical_list: D007477:Ions; D008670:Metals; D011994:Recombinant Proteins; D014866:Waste Products; D014990:Xylans; D043364:Endo-1,4-beta Xylanases
keywords: Catalysis improvement; Glycoside hydrolase family 10 (GH10); Hyperthermostable; Talaromyces leycettanus; Xylanase