Efficacious bioconversion of alginate/cellulose to value-added oligosaccharides by alginate-degrading GH5 endoglucanase from Trichoderma asperellum

被引：1

作者：

Zhuang, Huan ^{[1
]}

Zheng, Fengzhen ^{[2
]}

Zhang, Hengbin ^{[2
]}

Wang, Jiaqiang ^{[2
]}

Chen, Jun ^{[3
]}

机构：

[1] Zhejiang Univ, Childrens Hosp, Sch Med, Dept ENT & Head & Neck Surg, Hangzhou 310051, Zhejiang, Peoples R China

[2] Zhejiang Shuren Univ, Coll Biol & Environm Engn, Hangzhou 310021, Peoples R China

[3] Zhejiang Shuren Univ, Interdisciplinary Res Acad, Hangzhou 310021, Peoples R China

来源：

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES | 2024年 / 270卷

基金：

中国国家自然科学基金;

关键词：

Trichoderma asperellum; Endoglucanase; Bifunctional; Catalytic sites; Cellobiose; Unsaturated oligosaccharides; PICHIA-PASTORIS; PROCESSIVE ENDOGLUCANASE; ALPHA-GALACTOSIDASE; BROWN SEAWEED; PURIFICATION; LYASE; CELLULASE; ENZYME; SACCHARIFICATION; STRAIN;

D O I：

10.1016/j.ijbiomac.2024.131968

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 degradation of lignocellulosic biomass provides an eco-friendly approach to produce value-added macromolecules, e.g., bioactive polysaccharides. A novel acidophilic GH5 8-1,4-endoglucanase (termed TaCel5) from Trichoderma asperellum ND-1 was efficiently expressed in Komagataella phaffii (similar to 1.5-fold increase, 38.42 U/mL). TaCel5 displayed both endoglucanase (486.3 U/mg) and alginate lyase (359.5 U/mg) enzyme activities. It had optimal pH 3.0 and strong pH stability (exceed 86 % activity retained over pH range 3.0-5.0). 80 % activity (both endoglucanase and alginate lyase) was retained in the presence of 15 % ethanol or 3.42 M NaCl. Analysis of action mode revealed that hydrolytic activity of TaCel5 required at least three glucose (cellotriose) residues, yielding mainly cellobiose. Glu(241) and Glu(352) are essential catalytic residues, while Asp(106), Asp(277) and Asp(317) play auxiliary roles in cellulose degradation. TaCel5 displayed high hydrolysis efficiency for glucan and alginate substrates. ESI-MS analysis indicated that the enzymatic hydrolysates of alginate mainly contained disaccharides and heptasaccharides. This is the first detailed report of a bifunctional GH5 endoglucanase/alginate lyase enzyme from T. asperellum. Thus TaCel5 has strong potential in food and feed industries as a catalyst for bioconversion of cellulose- and alginate-containing waste materials into value-added products oligosaccharides, which was of great benefit both for the economy and environment.

引用

页数：12