Purification and characterization of a cell wall hydrolase encoded by the cw1A gene of Bacillus subtilis |
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| Authors: | Akio Kuroda Masafumi Imazeki Junichi Sekiguchi |
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| Institution: | Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, Nagano, Japan. |
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| Abstract: | 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. |
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| Keywords: | Cell wall hydroxylase Peptidoglycan Bacillus subtilis l-alanine amidase |
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