Penicillin G Amidase생산을 위한 재조합 대장균의 유가배양에 관한 연구
Fed-batch Culture of Recombinant E.coli for the Production of Penicillin G Amidase.
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Penicillin G amidase(PGA, benzylpenicillinaminohydrolase, EC 3.5.1.11) is industrially important enzyme which converts penicillinG 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.
Abstract
Penicillin G amidase(PGA, benzylpenicillinaminohydrolase, EC 3.5.1.11) is industrially important enzyme which converts penicillinG 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.
참고문헌 (24)
- Buttrick, P., "The regulation of heat shock protein expression: how, when and where", J. Mol. Cell. Cardiol, 41, pp.785~786 (2006)
- Carty, S. M., "Effect of cold shock on lipid A biosynthesis in Escherichia coli", J. Biol. Chem, 통권 : 274, pp.9677~9685 (1999)
- Cheng, S. W., "High-level production penicillin G acylase from Alcaligenes faecalis in recombinant Escherichia coli with optimization of carbon source", Enzyme. Microb. Technol, 41, pp.326~330 (2007)
- Choi, K. S., "Effects of sitedirected mutations on processing and activities of penicillin G acylase from Escherichia coli ATCC 11105", J. Bacteriol, 통권 : 174, pp.6270~6276 (1992)
- Deak, P. M., "Bioreactor cultivation of Escherichia coli for production of recombinant penicillin G amidase from Alcaligenes faecalis", Biotech. Lett, 25, pp.397~400 (2003)
- French, C., "Development of a simple method for the recovery of recombinant proteins from the Escherichia coli periplasm", Enzyme Microb. Technol, 19, pp.332~338 (1996)
- Fukushima, H., "Changes in membrane lipid composition during temperature adaptation by a thermotolerant strain of Tetrahymena pyriformis", Biochim. Biophys. Acta, 43, pp.165~179 (1976)
- Kheirolomoom, A., "Clarification of penicillin G acylase reaction mechanism", Process Biochem, 36, pp.1095~1101 (2001)
- KIM, JIN-HEE, "One-step Purification of Poly-His Tagged Penicillin G Acylase Expressed inE. coli", Journal of Microbiology and Biotechnology, 14(2), pp.231~236 (2004), 한국미생물·생명공학회
- Lamotte, D., "temperature profile in batch culture to increase the production of the recombinant UDPglucuronosyltransferase 2B4 in Escherichia coli", Process Biochem, 31, pp.235~241 (1996)
- John Chi-Wei Lan, "A Fermentation Strategy for Anti-MUC1 C595 Diabody Expression in Recombinant Escherichia coli", Biotechnology and Bioprocess Engineering, 11(5), pp.425~431 (2006), 한국생물공학회
- Oh, S. J., "Complete nucleotide sequence of the penicillin G acylase gene and the flanking regions, and its expression in Escherichia coli", Gene, 56, pp.87~97 (1987)
- Panoff, J. M., "Cold stress responces in mesophilic bacteria", Cryobiology, 36, pp.75~83 (1998)
- Park, Y. C., "Batch and fedbatch production of coenzyme Q(10) in recombinant Escherichia coli containing the decaprenyl diphosphate synthase gene from Gluconobacter suboxydans", Appl. Microbiol. Biotechnol, 67, pp.192~196 (2005)
- Pierce, J. J., "Factors determining more efficient large-scale release of a periplasmic enzyme from E. coli using lysozyme", J. Biotech, 58, pp.1~11 (1997)
- Rinas, U., "Glucose as a substrate in recombinant strain fermentation technology by-product formation, degradation and intracellular accumulation of recombinant protein", Appl. Microbiol. Biotechnol, 31, pp.163~167 (1989)
- Sambrook, J., "Molecular Cloning: A Laboratory Manual, 2nd ed" (1989), Cold Spring Harbor Laboratory Press
- Sobotková, L., "Development of a high-expression system for penicillin G acylase based on the recombinant Escherichia coli strain RE3(pKA18).", Enzyme and Microbial Technology, 19, pp.389~397 (1996)
- Strandberg, L,, "Batch and fed batch cultivations for the temperature induced production of a recombinant protein in Escherichia coli", Biotechnol. Lett, 13, pp.609~614 (1991)
- Vaara, M., "Outer membrane permeability barrier in Escherichia coli mutant that are defective in the late acyltransferase of lipid A biosynthsis", Antimicrob. Agents Chemother, 43, pp.1469~1462 (1999)
- Vorachek-Warren, M. K., "An Escherichia coli mutant lacking the cold shock-induced palmitoleoyltransferase of lipid A biosynthesis", J. Biol. Chem, 통권 : 277, pp.14186~14193 (2002)
- Wouters, J. A., "Changes in glycolytic activity of Lactococcus lactis induced by low temperature", Appl. Environ. Microbiol, 66, pp.3686~3691 (2000)
- Yang, Y., "High yield recombinant penicillin G amidase production and export into the growth medium using Bacillus megaterium", Microbial Cell Factories, 5, pp.36~50 (2006)
- Yim, S. C., "High-level secretory production of granulocyte-colony stimulating factor by fed-batch culture of recombinant Escherichia coli", Bioprocess and Biosystems Engineering, 24, pp.249~254 (2001)