Open AccessPurification and characterization of a cell wall hydrolase encoded by the cw1A gene of Bacillus subtilis
Author(s) -
Kuroda Akio,
Imazeki Masafumi,
Sekiguchi Junichi
Publication year - 1991
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1991.tb04703.x
Subject(s) - bacillus subtilis , biochemistry , hydrolase , escherichia coli , amidase , microbiology and biotechnology , molecular mass , enzyme , plasmid , cell wall , chemistry , zymography , biology , gene , bacteria , genetics
A cell wall hydrolase of Bacillus subtilis was prepared from Escherichia coli cells harboring a plasmid containing the B. subtilis cw1A gene and purified by hydroxyapatite column chromatography and HPLC through TSK‐gel G3000SW XL . 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.