期刊
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
卷 101, 期 18, 页码 6929-6939出版社
SPRINGER
DOI: 10.1007/s00253-017-8430-2
关键词
Xylanase; Cellulase; Fusion protein; Metagenome; Protein engineering
资金
- National Institute of Genetic Engineering and Biotechnology (NIGEB) [518, 578]
The camel rumen metagenome is an untapped source of glycoside hydrolases. In this study, novel genes encoding for a modular xylanase (XylC) and a cellulase (CelC) were isolated from a camel rumen metagenome and expressed in Escherichia coli BL21 (DE3). XylC with xylanase (Xyn), CBM, and carbohydrate esterase (CE) domains was characterized as a beta-1,4-endoxylanase with remarkable catalytic activity on oat-spelt xylan (K-cat = 2919 +/- 57 s(-1)). The implication of XylC's modular structure in its high catalytic activity was analyzed by truncation and fusion construction with CelC. The resulting fusions including Cel-CBM, Cel-CBM-CE, and Xyn-CBM-Cel showed remarkable enhancement in CMCase activity with K (cat) values of 742 +/- 12, 1289 +/- 34.5, and 2799 +/- 51 s(-1) compared to CelC with a K (cat) of 422 +/- 3.5 s(-1). It was also shown that the bifunctional Xyn-CBM-Cel with synergistic xylanase/cellulase activities was more efficient than XylC and CelC in hydrolysis of rice and barley straws.
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