Secretion of active-form Streptoverticillium mobaraense transglutaminase by Corynebacterium glutamicum:: Processing of the pro-transglutaminase by a cosecreted subtilisin-like protease from Streptomyces albogriseolus
Secretion of active-form Streptoverticillium mobaraense transglutaminase by Corynebacterium glutamicum:: Processing of the pro-transglutaminase by a cosecreted subtilisin-like protease from Streptomyces albogriseolus
复制标题
DOI：
10.1128/aem.69.1.358-366.2003
复制
发表时间：
2003-01-01
影响因子：
4.4
通讯作者：
Matsui, H
中科院分区：
文献类型：
--
作者：
Kikuchi, Y;Date, M;Matsui, H
莫巴拉链霉菌分泌的转谷氨酰胺酶是一种在食品工业中非常有用的酶。谷氨酸棒杆菌将谷氨酰胺转氨酶与谷氨酸杆菌细胞表面蛋白的启动子和信号肽偶联后，分泌出谷氨酰胺转氨酶的一个片段。我们分析了转谷氨酰胺酶基因的信号肽和原结构域，发现信号肽由31个氨基酸残基组成，原结构域由45个残基组成。利用转谷氨酰胺酶原结构域，谷氨酰胺转氨酶原能被谷氨酰胺转氨酶高效分泌，但没有酶活性。然而，当携带莫巴拉链霉菌转谷氨酰胺酶的质粒也编码SAM-P45时，白链霉菌的一种类似枯草杆菌的丝氨酸蛋白酶，与转谷氨酰胺酶原酶41-Ser的C侧结合的多肽被水解，转谷氨酰胺转氨酶原酶被转化为活性形式。我们的发现表明，谷氨酸杆菌具有作为工业规模蛋白质生产宿主的潜力。
The transglutaminase secreted by Streptoverticillium mobaraense is a useful enzyme in the food industry. A fragment of transglutaminase was secreted by Corynebacterium glutamicum when it was coupled on a plasmid to the promoter and signal peptide of a cell surface protein from C. glutamicum. We analyzed the signal peptide and the pro-domain of the transglutaminase gene and found that the signal peptide consists of 31 amino acid residues and the pro-domain consists of 45 residues. When the pro-domain of the transglutaminase was used, the pro-transglutaminase was secreted efficiently by C. glutamicum but had no enzymatic activity. However, when the plasmid carrying the S. mobaraense transglutaminase also encoded SAM-P45, a subtilisin-like serine protease derived from Streptomyces albogriseolus, the peptide bond to the C side of 41-Ser of the pro-transgiutaminase was hydrolyzed, and the pro-transglutaminase was converted to an active form. Our findings suggest that C. glutamicum has potential as a host for industrial-scale protein production.