School of Pharmaceutical Science, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, China.
State Key Lab of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, No. 1 West Beichen Road, Chaoyang District, Beijing, 100101, China.
Bioprocess Biosyst Eng. 2018 Oct;41(10):1437-1447. doi: 10.1007/s00449-018-1972-6. Epub 2018 Jun 23.
A novel alkaline protease (named AprV) gene from Vibrio sp. DA1-1 was cloned and expressed in Escherichia coli BL21 (DE3) pLysS. The sequence analysis showed the highest homology of 68% with the characterized protease from Alkalimonas collagenimarina AC40. The recombinant AprV was purified with the molecular weight of 28 kDa. The optimum temperature and pH were determined to be 55 °C and 10.0, respectively. The enzyme activity was slightly enhanced by Ca, Mg, Zn, Ba, and, however, was highly inhibited by Sn and EDTA. The AprV was stable in the presence of some surfactants and oxidizing agents, such as 1% Tween 20-80, 1% JFC-2, and 5% JFC-2. Casein was found to be the ideal substrate with specific activity of 1139 U/mg. Moreover, we found that AprV (10,000 U), together with commercial detergent, could completely remove the blood on the cotton. Furthermore, AprV also demonstrated dehairing activity on goat and bull skin. These results indicated that the alkaline protease AprV might be a potential candidate for applications in the detergent and leather industries.
从 Vibrio sp. DA1-1 中克隆并在 Escherichia coli BL21 (DE3) pLysS 中表达了一种新型碱性蛋白酶（命名为 AprV）基因。序列分析表明，其与 Alkalimonas collagenimarina AC40 中鉴定的蛋白酶的同源性最高，为 68%。重组 AprV 的分子量为 28 kDa，通过亲和层析进行纯化。确定最佳温度和 pH 值分别为 55°C 和 10.0。该酶的活性在一定程度上受到 Ca、Mg、Zn、Ba 的增强，但受到 Sn 和 EDTA 的强烈抑制。AprV 在存在一些表面活性剂和氧化剂（如 1%Tween 20-80、1%JFC-2 和 5%JFC-2）时稳定。酪蛋白被发现是具有 1139 U/mg 比活的理想底物。此外，我们发现 10000 U AprV 与商业洗涤剂一起可以完全去除棉上的血迹。此外，AprV 对山羊皮和牛皮也具有脱毛活性。这些结果表明，碱性蛋白酶 AprV 可能是在洗涤剂和制革工业中应用的潜在候选物。
