Open AccessGene cloning and characterization of a second alanine racemase from Bacillus subtilis encoded by yncD
Author(s) -
Pierce Kara J.,
Salifu Samson P.,
Tangney Martin
Publication year - 2008
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.1574-6968.2008.01151.x
Subject(s) - bacillus subtilis , auxotrophy , escherichia coli , biology , alanine , plasmid , gene , biochemistry , mutant , microbiology and biotechnology , molecular cloning , bacteria , amino acid , genetics , peptide sequence
Alanine racemase activity was investigated in Bacillus subtilis . A putative second alanine racemase gene ( yncD ) was cloned in parallel with the previously identified alanine racemase gene, dal . Each of the B. subtilis genes, dal and yncD complemented the Escherichia coli Alr − DadX − double mutant alanine auxotrophic strain MB2159 in vivo , restoring the prototrophic phenotype. Alanine racemase activity was also detected in vitro in cell‐free extracts prepared from cultures of E. coli MB2159 harboring plasmids expressing either of the cloned B. subtilis genes and preliminary characterization of enzyme activity is presented.