一种新型酪氨酸酶的克隆、鉴定及其在卡氏链霉菌SC-1中的过表达以提高黑色素产量
Cloning and identification of a novel tyrosinase and its overexpression in Streptomyces kathirae SC-1 for enhancing melanin production.
作者信息
Guo Jing, Rao Zhiming, Yang Taowei, Man Zaiwei, Xu Meijuan, Zhang Xian, Yang Shang-Tian
机构信息
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu Province 214122, P. R. China The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, P. R. China.
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu Province 214122, P. R. China The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, P. R. China
出版信息
FEMS Microbiol Lett. 2015 Apr;362(8):fnv041. doi: 10.1093/femsle/fnv041. Epub 2015 Mar 10.
A 30-kDa novel tyrosinase was purified to homogeneity. The Km for L-Dopa and L-tyrosine were determined as 0.42 and 0.25 mM. The 1231 bp (base pair) melC gene and its 167 bp promoter Pskmel were obtained by thermal asymmetric interlaced polymerase chain reaction based on the amino acids fragment obtained from MS results of the purified enzyme. The protein sequence of tyrosinase shows maximum identity (84%) to tyrosinase from Streptomyces galbus. The melC was introduced into S. kathirae. The melanin production and the transcriptional level of melC in recombinant S. kathirae [pIJPskmelmelC] were about 2.1-fold and 2-fold higher than the wild-type strain, respectively. The melanin concentration was maximized at 28.8 g L(-1).
一种30 kDa的新型酪氨酸酶被纯化至同质。L-多巴和L-酪氨酸的米氏常数分别测定为0.42和0.25 mM。基于纯化酶的质谱结果获得的氨基酸片段，通过热不对称交错聚合酶链反应获得了1231 bp（碱基对）的melC基因及其167 bp的启动子Pskmel。酪氨酸酶的蛋白质序列与来自加尔布斯链霉菌的酪氨酸酶具有最高84%的同一性。将melC导入卡氏链霉菌。重组卡氏链霉菌[pIJPskmelmelC]中黑色素的产生和melC的转录水平分别比野生型菌株高约2.1倍和2倍。黑色素浓度在28.8 g L(-1)时达到最大值。
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