High expression of the penicillin G acylase gene (pac) from Bacillus megaterium UN1 in its own pac minus mutant.
By marker exchange mutagenesis, Bacillus megaterium strain UN-1 (Bm-UN1) was used to prepare a mutant strain B. megaterium UN-cat (Bm-UNcat) lacking the penicillin G acylase gene (pac). The pac gene from Bm-UN1 was subcloned into pTF6 and the resultant plasmid, pBA402, was introduced into Bm-UNcat and Bacillus subtilis. Bm-UNcat harbouring pBA402 produced high penicillin G acylase (PAC) activity of 13.7, 19.5 and 20.4 U ml(-1) at 24, 36 and 48 h of culture, respectively. This was two- to fivefold higher than PAC produced by B. subtilis harbouring pBA402 and about 20-fold higher than PAC produced by the parent strain, Bm-UN1.
Panbangred
W
W
Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok, Thailand. grwpb@mahidol.ac.th
Weeradechapon
K
K
Udomvaraphant
S
S
Fujiyama
K
K
Meevootisom
V
V
D000595:Amino Acid Sequence; D001410:Bacillus megaterium / Q000201:enzymology / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D015870:Gene Expression; D005798:Genes, Bacterial; D009154:Mutation; D010399:Penicillin Amidase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014169:Transformation, Bacterial
D010399:Penicillin Amidase