Purification and characterization of a cell wall hydrolase encoded by the cwlA gene of Bacillus subtilis.
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.
Kuroda
A
A
Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, Nagano, Japan.
Imazeki
M
M
Sekiguchi
J
J
D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D002473:Cell Wall / Q000201:enzymology; D004792:Enzyme Precursors / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D006867:Hydrolases / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification*; D009994:Osmolar Concentration
D004792:Enzyme Precursors; D006867:Hydrolases