A new locus affects cell motility, cellulose binding, and degradation by Cytophaga hutchinsonii

被引:0
作者
Xiaofei Ji
Yuanxi Xu
Cong Zhang
Ning Chen
Xuemei Lu
机构
[1] Shandong University,State Key Laboratory of Microbial Technology, College of Life Science
来源
Applied Microbiology and Biotechnology | 2012年 / 96卷
关键词
Gene complementation; Cellulose binding protein; Cellulose degradation;
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摘要
Cytophaga hutchinsonii is a Gram-negative gliding bacterium, which can rapidly degrade crystalline cellulose via a novel strategy without any recognizable processive cellulases. Its mechanism of cellulose binding and degradation is still a mystery. In this study, the mutagenesis of C. hutchinsonii with the mariner-based transposon HimarEm3 and gene complementation with the oriC-based plasmid carrying the antibiotic resistance gene cfxA or tetQ were reported for the first time to provide valuable tools for mutagenesis and genetic manipulation of the bacterium. Mutant A-4 with a transposon mutation in gene CHU_0134, which encodes a putative thiol-disulfide isomerase exhibits defects in cell motility and cellulose degradation. The cellulose binding ability of A-4 was only half of that of the wild-type strain, while the endo-cellulase activity of the cell-free supernatants and on the intact cell surface of A-4 decreased by 40 %. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of proteins binding to cellulose in the outer membrane showed that most of them were significantly decreased or disappeared in A-4 including some Gld proteins and hypothetical proteins, indicating that these proteins might play an important role in cell motility and cellulose binding and degradation by the bacterium.
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页码:161 / 170
页数:9
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