A small periplasmic protein essential for Cytophaga hutchinsonii cellulose digestion
A small periplasmic protein essential for Cytophaga hutchinsonii cellulose digestion
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哈钦森噬细胞纤维素消化所必需的小周质蛋白
DOI：
10.1007/s00253-015-7204-y
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发表时间：
2016
影响因子：
5
通讯作者：
Liu Weifeng
中科院分区：
文献类型：
--
作者：
Yang Tengteng;Bu Xuliang;Han Qingqing;Wang Xia;Zhou Hong;Chen Guanjun;Zhang Weixin;Liu Weifeng
哈氏细胞噬菌是一种广泛分布于土壤中的降解纤维素的细菌。c。然而，hutchinsoni实现纤维素消化的机制在很大程度上仍然未知。本研究通过转座子诱变获得了以滤纸或Avicel为唯一碳源存在缺陷的aC. hutchinson1突变体。中断的基因位点CHU_2981编码一种假设的蛋白质，只有130个氨基酸。与c端绿色荧光蛋白（GFP）融合的CHU_2981细胞分离和western blot检测表明，CHU_2981位于外周质。hu_2981突变体细胞在Avicel上表现出明显的生长缺陷，而在葡萄糖和纤维素二糖上则没有。与野生型细胞相比，CHU_2981的缺失也导致了集落扩展和单个细胞运动的显著缺陷。进一步分析表明，hu_2981突变体细胞的纤维素吸收外膜蛋白谱与野生型细胞不同，蛋白的种类和数量明显减少。结果表明，外质非纤维素水解蛋白CHU_2981在纤维素利用和细胞运动中都起着重要作用，可能参与了外膜蛋白的适当产生。
Cytophaga hutchinsoniiis a gliding cellulolytic bacterium that is ubiquitously distributed in soil. The mechanism by whichC. hutchinsoniiachieves cellulose digestion, however, is still largely unknown. In this study, we obtained aC. hutchinsoniimutant that was defective in utilizing filter paper or Avicel as the sole carbon source by transposon mutagenesis. The interrupted gene locus,CHU_2981, encodes a hypothetical protein with only 130 amino acids. Cell fractionation and western blot detection of CHU_2981 fused with a C-terminal green fluorescence protein (GFP) indicated that CHU_2981 is located in the periplasm. TheCHU_2981-disrupted mutant cells exhibited a significant growth defect on Avicel but not on glucose and cellobiose. The absence of CHU_2981 also resulted in a significant defect in colony spreading and individual cell motility compared to wild-type cells. Further analysis demonstrated that theCHU_2981-disrupted mutant cells exhibited a different profile of cellulose-absorbed outer membrane proteins from that of wild-type cells, in which protein varieties and amounts were markedly decreased. Our results showed that CHU_2981, the periplasmic non-cellulolytic protein, plays an important role in both cellulose utilization and cell motility probably by being involved in the appropriate production of outer membrane proteins.