Cloning, Expression, and Transglycosylation Reaction of Paenibacillus sp. Strain W-61 Xylanase 1
Cloning, Expression, and Transglycosylation Reaction of Paenibacillus sp. Strain W-61 Xylanase 1
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DOI：
10.1271/bbb.70622
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发表时间：
2008-04
期刊：
影响因子：
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通讯作者：
Seiji Watanabe;D. N. Viet;J. Kaneko;Y. Kamio;S. Yoshida
中科院分区：
文献类型：
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作者：
Seiji Watanabe;D. N. Viet;J. Kaneko;Y. Kamio;S. Yoshida
从大肠杆菌中克隆了类芽孢杆菌（Paenibacillus sp.）W-61菌株木聚糖酶基因xyn 1。xyn 1编码211个氨基酸残基，其中信号肽28个氨基酸残基。Xyn 1成熟蛋白的氨基酸序列与AeromonascausME-1、Paenibacillussp.和嗜热脂肪芽孢杆菌。重组Xyn 1的理化性质与完整的Xyn 1非常相似，除了分子量。利用荧光辅助糖电泳（FACE）研究了rXyn 1产生的低聚木糖的模式。与木五糖相比，rXyn 1对木六糖的降解率显著提高。木六糖有一个单一的优先点裂解rXyn 1。根据低聚木糖的作用模式，亚位点的数量估计为6个。催化中心位于非还原端第3和第4个亚中心之间。
A xylanase gene, xyn1, which encodes Paenibacillus sp. strain W-61 xylanase 1 (Xyn1), was cloned in Escherichia coli. xyn1 encodes 211 amino acid residues, including 28 amino acid residues of a signal peptide. The deduced amino acid sequence of the mature Xyn1 showed 95.7%, 84.0%, and 83.7% identity to family 11 xylanases of Aeromonas caviae ME-1, Paenibacillus sp., and Bacillus stearothermophilus respectively. The physico-chemical properties of recombinant Xyn1 were very similar to those of intact Xyn1, except for the molecular mass. The pattern of xylooligosaccharides generated by rXyn1 was investigated by fluorophore-assisted carbohydrate electrophoresis (FACE). The degradation rate of xylohexaose by rXyn1 increased markedly as compared with that of xylopentaose. Xylohexaose had a single preferential point of cleavage by rXyn1. On the basis of the pattern of action of xylooligosaccharides, the number of subsites was estimated to be six. The catalytic site was located between the third and the fourth subsites from non-reducing end.