Purification and molecular characterization of subtilisin-like alkaline protease BPP-A from Bacillus pumilus strain MS-1
Purification and molecular characterization of subtilisin-like alkaline protease BPP-A from Bacillus pumilus strain MS-1
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DOI：
10.1111/j.1472-765x.2005.01851.x
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发表时间：
2006-03-01
影响因子：
2.4
通讯作者：
Tomizuka, N
中科院分区：
文献类型：
--
作者：
Miyaji, T;Otta, Y;Tomizuka, N
目标：本研究通过筛选玉米醇溶蛋白降解菌，以获得玉米醇溶蛋白降解蛋白酶。方法和结果：通过在玉米醇溶蛋白平板上形成透明带的方法筛选土壤细菌。对一株玉米醇溶蛋白降解菌的生物学特性分析表明，该菌属于短小芽孢杆菌（Bacilluspumilus），命名为MS-1.该菌株产生两种不同类型的胞外蛋白酶，BPP-A和BPP-B。在这项研究中，我们纯化和表征BPP-A，因为它表现出比BPP-B更高的水解玉米醇溶蛋白的能力。以酪蛋白为底物时，BPP-A的最适pH为11。0.在BPP-A中，玉米醇溶蛋白比酪蛋白更适合作为底物。0，而在pH 11（.）0. bppA基因编码一个由383个氨基酸组成的BPP-A前体，成熟BPP-A由275个氨基酸组成。结论：BPP-A属于枯草杆菌蛋白酶家族。pumilus产生两种不同类型的胞外蛋白酶BPP-A和BPP-B。BPP-A具有在极端碱性条件下水解玉米醇溶蛋白的能力。本研究的意义和影响：本研究首次报道了玉米醇溶蛋白降解微生物的筛选。枯草杆菌蛋白酶样蛋白酶BPP-A有可能用作生产玉米醇溶蛋白水解产物的工业酶。
Aims: The present study was conducted by screening zein-degrading bacteria in an attempt to obtain zein- degrading protease.Methods and Results: Soil bacteria were screened by formation of a clear zone on zein plates. Characterization of a zein- degrading bacterium indicated a taxonomic affiliation to Bacillus pumilus, and was named MS-1 strain. The strain produced two different types of extracellular proteases, BPP-A and BPP-B. In this study, we purified and characterized BPP-A because it exhibited a higher ability to hydrolyze zein than BPP-B. When casein was used as the substrate, the optimal pH for BPP-A was 11(.)0. In BPP-A, zein was better substrate than casein at pH 13(.)0, whereas casein was better one than zein at pH 11(.)0. The bppA gene encoded a 383-amino acid pre-pro form of BPP-A, and mature BPP-A contained 275 amino acid residues. It was concluded that BPP-A belonged to the subtilisin family.Conclusion: A zein-degrading bacterium assigned to B. pumilus produced two different types of extracellular proteases, BPP-A and BPP-B. BPP-A exhibited an ability to hydrolyze zein in an extreme alkaline condition.Significance and Impact of the Study: This is a first report on screening for zein-degrading micro-organisms. The subtilisin-like protease BPP-A is possible to utilize as an industrial enzyme for the production of zein hydrolysates.