Expression and characterization of a novel halophilic GH10 β-1,4-xylanase from Trichoderma asperellum ND-1 and its synergism with a commercial α-L-arabinofuranosidase on arabinoxylan degradation

被引:5
作者
Zheng, Fengzhen [1 ]
Zhang, Hengbin [1 ]
Wang, Jiaqiang [1 ]
Chen, Jun [2 ]
Zhuang, Huan [3 ]
Basit, Abdul [4 ]
机构
[1] Zhejiang Shuren Univ, Coll Biol & Environm Engn, Hangzhou 310021, Peoples R China
[2] Zhejiang Shuren Univ, Interdisciplinary Res Acad, Hangzhou 310021, Peoples R China
[3] Zhejiang Univ, Dept ENT & Head & Neck Surg, Sch Med, Childrens Hosp, Hangzhou 310051, Zhejiang, Peoples R China
[4] Univ Jhang, Dept Microbiol, Jhang 35200, Pakistan
基金
中国国家自然科学基金;
关键词
Trichoderma asperellum; Halophilic xylanase; Cleavage pattern; Catalytic sites; Synergistic action; PICHIA-PASTORIS; BIOCHEMICAL-CHARACTERIZATION; ENZYMATIC CHARACTERIZATION; THERMOSTABLE XYLANASE; ACTIVE-SITE; PURIFICATION; CLONING; HYDROLYSIS; TOLERANT; PROTEIN;
D O I
10.1016/j.ijbiomac.2024.136885
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Enzymatic hydrolysis of arabinoxylan is of cost-effective strategy to yield valuable macromolecules, e.g., xylooligosaccharides (XOS). A novel halophilic GH10 xylanase (TaXYL10) from Trichoderma asperellum ND-1 was over-expressed in Pichia pastoris and migrated as a single band (similar to 36 kDa) in SDS-PAGE. TaXYL10 displayed >80 % activity in the presence of 4.28 M NaCl and 10 % ethanol. Moreover, TaXYL10 exhibited optimal activity at pH 6.0 and 55 degrees C, and remarkable pH stability (>80 % activity at pH 4.0-6.0). K+ and Al3+ could remarkably promote TaXYL10 activity, while the presence of 10 mM Fe2+, Zn2+, Cu2+ and Fe3+ decreased its activity. TaXYL10 possesses the highest catalytic activity towards beechwood xylan. TLC analysis revealed that it could rapidly degrade xylan and XOS with DP >= 3, yielding xylotriose and xylobiose. Site-directed mutagenesis indicated that Glu(154) and Glu(259) are crucial active residues for TaXYL10, while Asp(295) and Glu(69) played auxiliary roles in xylan hydrolysis. Additionally, TaXYL10 acted cooperatively with a commercial alpha-L-arabinofuranosidase (AnAra) towards arabinoxylan degradation (583.5 mu g/mL), a greater synergy degree of 1.79 was obtained after optimizing enzymatic ratios. This work not only expands the diversity of Trichoderma GH10 xylanases, but also reveals the promising potential of TaXYL10 in various industrial applications.
引用
收藏
页数:9
相关论文
共 57 条
[1]   From lignocellulosic residues to market: Production and commercial potential of xylooligosaccharides [J].
Amorim, Claudia ;
Silverio, Sara C. ;
Prather, Kristala L. J. ;
Rodrigues, Ligia R. .
BIOTECHNOLOGY ADVANCES, 2019, 37 (07)
[2]   Cloning, expression and characterization of a novel salt-tolerant xylanase from Bacillus sp SN5 [J].
Bai, Wenqin ;
Xue, Yanfen ;
Zhou, Cheng ;
Ma, Yanhe .
BIOTECHNOLOGY LETTERS, 2012, 34 (11) :2093-2099
[3]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[4]   A novel thermostable and halotolerant xylanase from Colletotrichum graminicola [J].
Carli S. ;
Meleiro L.P. ;
Rosa J.C. ;
Moraes L.A.B. ;
Jorge J.A. ;
Masui D.C. ;
Furriel R.P.M. .
Journal of Molecular Catalysis B: Enzymatic, 2016, 133 :S508-S517
[5]   Molecular characterization of a cold-active recombinant xylanase from Flavobacterium johnsoniae and its applicability in xylan hydrolysis [J].
Chen, Shicheng ;
Kaufman, Michael G. ;
Miazgowicz, Kerri L. ;
Bagdasarian, Michael ;
Walker, Edward D. .
BIORESOURCE TECHNOLOGY, 2013, 128 :145-155
[6]   Biochemical analyses of a novel thermostable GH5 endo β-1,4-mannanase with minor β-1,4-glucosidic cleavage activity from Bacillus sp. KW1 and its synergism with a commercial α-galactosidase on galactomannan hydrolysis [J].
Chen, Xi ;
Wang, Xinhai ;
Liu, Yun ;
Zhang, Ruiqin ;
Zhang, Liang ;
Zhan, Ruoting ;
Wang, Sidi ;
Wang, Kui .
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES, 2021, 166 :778-788
[7]   Insights into the roles of non-catalytic residues in the active site of a GH10 xylanase with activity on cellulose [J].
Chu, Yindi ;
Tu, Tao ;
Penttinen, Leena ;
Xue, Xianli ;
Wang, Xiaoyu ;
Yi, Zhuolin ;
Gong, Li ;
Rouvinen, Juha ;
Luo, Huiying ;
Hakulinen, Nina ;
Yao, Bin ;
Su, Xiaoyun .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2017, 292 (47) :19315-19327
[8]   Xylanases, xylanase families and extremophilic xylanases [J].
Collins, T ;
Gerday, C ;
Feller, G .
FEMS MICROBIOLOGY REVIEWS, 2005, 29 (01) :3-23
[9]   Cloning and constitutive expression of His-tagged xylanase GH 11 from Penicillium occitanis Pol6 in Pichia pastoris X33: Purification and characterization [J].
Driss, Dorra ;
Bhiri, Fatma ;
ghorbel, Raoudha ;
Chaabouni, Semia Ellouz .
PROTEIN EXPRESSION AND PURIFICATION, 2012, 83 (01) :8-14
[10]   Cloning and expression of a xylanase xynB from Aspergillus niger IA-001 in Pichia pastoris [J].
Fang, Wei ;
Gao, He ;
Cao, Yunhe ;
Shan, Anshan .
JOURNAL OF BASIC MICROBIOLOGY, 2014, 54 :S190-S199