The physical role of the proteases from a marine bacterium, Alteromonas sp. Strain 0-7

海洋细菌交替单胞菌蛋白酶的物理作用。

• 批准号: 15580069
• 负责人: MIYAMOTO Katsushiro
• 金额: $ 2.3万
• 依托单位: Osaka University of Pharmaceutical Sciences
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15580069/
• 关键词: marine bacteria; response regulator; histidine kinase; chitin; protease; ArcA; ArcB

Within the extracellularproteases from the marine bacterium, Alteromonassp.strain O-7, AprIV is involved in the chitinolytic system. AprIV and chitinases (ChiA.ChiB,ChiC, and ChiD) were induced in the presence of N-acetylglucosamine (GlcNAc). To clarify the regulation of AprIV at the gene level, N-terminal amino acid sequence of the protein binding the upstream region of aprIV gene was determined. The result suggest that the regulatory protein is an anoxic redox control protein (ArcA), which is classified into the response regulator of ArcB/ArcA two-component signal transduction system. Thus, we cloned the areA and arcB gene of the strain, and expression plasmids ArcA and ArcB lacking the N-terminal membrane binding region (ArcBAN) were constructed by using the GST fusion protein expression vector pGEX-6P-1. The ArcA protein was transphosphorylated by the GstArcBΔN protein as a sensor kinase. Gel retardation assay and BIAcore analysis demonstrated that phosphorylated ArcA (ArcA-P) more tightly bound to the upstream region of aprIV and chiD genes compared to those of chiA,chiB,chiC, and cpb1 genes. These results suggest that ArcB/ArcA two-component signal transduction system regulates the expression of genes involved in chitin degradation system of the strain.

在来自海洋细菌的细胞外propece中，在存在N-乙酰基葡萄糖（GlcNAC）的情况下，诱导了Alteromonasp.Strain O-7 SES（Chia.chib，Chic和Chid）。调节蛋白是一种缺氧的氧化还原对照蛋白（ARCA），该蛋白被分类为组成型信号转导系统，因此，我们将菌株SARCA和ARCB的ARCB基因克隆为ARCB。使用GSTT融合蛋白表达载体PGEX-6P-1构建（Arcban）。对于CHIA，CHIB和CPB1基因的结果。