Isolation and biochemical characterization of an endo-1,3-beta-glucanase from Streptomyces sioyaensis containing a C-terminal family 6 carbohydrate-binding module that binds to 1,3-beta-glucan

Microbiology
Tang Yao HongMenghsiao Meng

Abstract

A gene encoding 1,3-beta-glucanase was isolated from Streptomyces sioyaensis based on an activity plate assay. Analysis of the deduced amino acid sequence of the gene revealed that the matured 1,3-beta-glucanase has two functional domains separated by a stretch of nine glycine residues. The N-terminal domain shares sequence similarity with bacterial endo-1,3-beta-glucanases classified in glycosyl hydrolase family 16 (GHF 16), while the C-terminal domain is a putative carbohydrate-binding module (CBM) grouped into CBM family 6. To characterize the function of each domain, both the full-length and the CBM-truncated versions of the protein were expressed in Escherichia coli and purified to homogeneity. Biochemical data suggest that the glycosyl hydrolase domain preferentially catalyses the hydrolysis of glucans with 1,3-beta linkage, and has an endolytic mode of action. Binding assay indicated that the C-terminal CBM binds to various insoluble beta-glucans (1,3-, 1,3-1,4- and 1,4- linkages) but not to xylan, a primary binding target for most members of CBM family 6. The full-length and the CBM-truncated proteins had similar specific activity (units per mol of hydrolase domain) on soluble 1,3-beta-glucan, whereas the former had muc...Continue Reading

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Citations

Sep 24, 2015·Applied Microbiology and Biotechnology·Tohru KobayashiShigeru Deguchi
Jan 19, 2010·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Jung Min SongYoun-Ho Lee
Aug 6, 2015·Applied Biochemistry and Biotechnology·Thabata Maria AlvarezFabio Marcio Squina
Feb 7, 2015·Protein Expression and Purification·Lorène BelvalJean-François Chich
Aug 26, 2014·Frontiers in Chemistry·Mohammad M RahmanTakao Ojima
Oct 20, 2012·Protein Engineering, Design & Selection : PEDS·Wei-Chun LiuLie-Fen Shyur
Dec 21, 2010·Applied and Environmental Microbiology·V BlättelH König
Sep 20, 2008·Applied Microbiology and Biotechnology·Yueh-Mei ChengMenghsiao Meng
Jun 30, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Qiulan WuXiangru Liao
Nov 23, 2019·Applied Microbiology and Biotechnology·Ahmet H BadurChristopher V Rao
Jul 15, 2020·International Journal of Biological Macromolecules·Roza V UsoltsevaSvetlana P Ermakova

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