|
KMID : 1234420080360040314
|
|
Korean Journal of Microbiololgy and Biotechnology
2008 Volume.36 No. 4 p.314 ~ p.319
|
|
|
Fed-batch Culture of Recombinant E.coli for the Production of Penicillin G Amidase
|
|
Lee Sang-Mahn
|
|
|
Abstract
|
|
|
|
Penicillin G amidase (PGA, benzylpenicillinaminohydrolase, EC 3.5.1.11) is industrially important enzyme which converts penicillin G to 6-aminopenicillanic acid (6-APA) and phenylacetic acid (PAA). The PGA in E. coli ATCC 11105 is secreted into the periplasm after removing signal sequences and becomes heterodimer which composed of two subunits, small subunit (24 kDa) and large subunit (65 kDa). In this study, the PGA gene was obtained from E. coli ATCC 11105 using PCR (polymerase chain reaction) technique. The active PGA was successfully secreated into periplasm in E. coli BL2 1(DE3) harboring pET-pga plasmid. The optimized fed-batch fermentation, consisting of a three-step shift of culture temperature from 37oC to 22oC, gave a productivity of 19.6 U/mL with a cell growth of 62 O.D. at 600 nm.
|
|
|
KEYWORD
|
|
|
Penicillin G amidase, recombinant E.coli, fed-batch culture
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|