枯草芽孢杆菌cwlA基因编码的一种细胞壁水解酶的纯化与特性分析
Purification and characterization of a cell wall hydrolase encoded by the cwlA gene of Bacillus subtilis.
作者信息
Kuroda A, Imazeki M, Sekiguchi J
机构信息
Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, Nagano, Japan.
出版信息
FEMS Microbiol Lett. 1991 Jun 1;65(1):9-13. doi: 10.1016/0378-1097(91)90462-j.
A cell wall hydrolase of Bacillus subtilis was prepared from Escherichia coli cells harboring a plasmid containing the B. subtilis cwlA gene and purified by hydroxyapatite column chromatography and HPLC through TSK-gel G3000SWXL. In contrast to the molecular mass of 29,919 Da deduced from its nucleotide sequence, the purified CWLA is a 23 kDa protein. Characterization of the specific substrate bond cleaved by CWLA indicated the enzyme is an N-acetylmuramyl-L-alanine amidase. A 32-kDa precursor protein was detected on zymography of a crude cell homogenate. Some of the enzymatic properties of CWLA are also described.
从携带含有枯草芽孢杆菌cwlA基因的质粒的大肠杆菌细胞中制备了枯草芽孢杆菌的一种细胞壁水解酶，并通过羟基磷灰石柱色谱法和TSK-gel G3000SWXL高效液相色谱法进行纯化。与根据其核苷酸序列推导的29,919 Da的分子量相反，纯化后的CWLA是一种23 kDa的蛋白质。对CWLA切割的特定底物键的表征表明该酶是一种N-乙酰胞壁酰-L-丙氨酸酰胺酶。在粗细胞匀浆的酶谱分析中检测到一种32 kDa的前体蛋白。还描述了CWLA的一些酶学性质。
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