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. 1989 Jun;55(6):1351–1356. doi: 10.1128/aem.55.6.1351-1356.1989

Expression of the Arthrobacter viscosus penicillin G acylase gene in Escherichia coli and Bacillus subtilis.

H Ohashi 1, Y Katsuta 1, M Nagashima 1, T Kamei 1, M Yano 1
PMCID: PMC202870  PMID: 2504107

Abstract

The penicillin G acylase gene cloned from Arthrobacter viscosus 8895GU was subcloned into vectors, and the recombinant plasmids were transferred into Escherichia coli or Bacillus subtilis. Both E. coli and B. subtilis transformants expressed the A. viscosus penicillin G acylase. The enzyme activity was found in the intracellular portion of the E. coli transformants or in the cultured medium of the B. subtilis transformants. Penicillin G acylase production in the B. subtilis transformants was 7.2 times higher than that in the parent A. viscosus. The A. viscosus penicillin G acylase was induced by phenylacetic acid in A. viscosus, whereas the enzyme was produced constitutively in both the E. coli and B. subtilis transformants carrying the A. viscosus penicillin G acylase gene.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Appleyard R K. Segregation of New Lysogenic Types during Growth of a Doubly Lysogenic Strain Derived from Escherichia Coli K12. Genetics. 1954 Jul;39(4):440–452. doi: 10.1093/genetics/39.4.440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  4. Daumy G. O., McColl A. S., Apostolakos D. Repression of penicillin G acylase of Proteus rettgeri by tricarboxylic acid cycle intermediates. J Bacteriol. 1982 Oct;152(1):104–110. doi: 10.1128/jb.152.1.104-110.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Daumy G. O., Williams J. A., McColl A. S., Zuzel T. J., Danley D. Expression and regulation of the penicillin G acylase gene from Proteus rettgeri cloned in Escherichia coli. J Bacteriol. 1986 Oct;168(1):431–433. doi: 10.1128/jb.168.1.431-433.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. GAREN A., LEVINTHAL C. A fine-structure genetic and chemical study of the enzyme alkaline phosphatase of E. coli. I. Purification and characterization of alkaline phosphatase. Biochim Biophys Acta. 1960 Mar 11;38:470–483. doi: 10.1016/0006-3002(60)91282-8. [DOI] [PubMed] [Google Scholar]
  7. Gang D. M., Shaikh K. Regulation of penicillin acylase in Escherichia coli by cyclic AMP. Biochim Biophys Acta. 1976 Feb 18;425(1):110–114. doi: 10.1016/0005-2787(76)90220-3. [DOI] [PubMed] [Google Scholar]
  8. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  9. Inoue M., Itoh J., Mitsuhashi S. pMS76, a plasmid capable of amplification by treatment with chloramphenicol. Plasmid. 1983 Jan;9(1):86–97. doi: 10.1016/0147-619x(83)90033-1. [DOI] [PubMed] [Google Scholar]
  10. Levitov M. M., Klapovskaia K. I., Kleiner G. I. Indutsirovannyi biosintez atsilazy Escherichia coli. Mikrobiologiia. 1967 Sep-Oct;36(5):912–917. [PubMed] [Google Scholar]
  11. Marrelli L. P. Colorimetric method for determination of 7-aminocephalosporanic acid (7-ACA) and related compounds. J Pharm Sci. 1968 Dec;57(12):2172–2173. doi: 10.1002/jps.2600571234. [DOI] [PubMed] [Google Scholar]
  12. Matsuda A., Komatsu K. I. Molecular cloning and structure of the gene for 7 beta-(4-carboxybutanamido)cephalosporanic acid acylase from a Pseudomonas strain. J Bacteriol. 1985 Sep;163(3):1222–1228. doi: 10.1128/jb.163.3.1222-1228.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Meevootisom V., Somsuk P., Prachaktam R., Flegel T. W. Simple screening method for isolation of penicillin acylase-producing bacteria. Appl Environ Microbiol. 1983 Nov;46(5):1227–1229. doi: 10.1128/aem.46.5.1227-1229.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Morrison D. A. Transformation and preservation of competent bacterial cells by freezing. Methods Enzymol. 1979;68:326–331. doi: 10.1016/0076-6879(79)68023-0. [DOI] [PubMed] [Google Scholar]
  15. Ohashi H., Katsuta Y., Hashizume T., Abe S. N., Kajiura H., Hattori H., Kamei T., Yano M. Molecular cloning of the penicillin G acylase gene from Arthrobacter viscosus. Appl Environ Microbiol. 1988 Nov;54(11):2603–2607. doi: 10.1128/aem.54.11.2603-2607.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Olsson A., Hagström T., Nilsson B., Uhlén M., Gatenbeck S. Molecular cloning of Bacillus sphaericus penicillin V amidase gene and its expression in Escherichia coli and Bacillus subtilis. Appl Environ Microbiol. 1985 May;49(5):1084–1089. doi: 10.1128/aem.49.5.1084-1089.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sadaie Y., Kada T. Formation of competent Bacillus subtilis cells. J Bacteriol. 1983 Feb;153(2):813–821. doi: 10.1128/jb.153.2.813-821.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Savidge T. A., Cole M. Penicillin acylase (bacterial). Methods Enzymol. 1975;43:705–721. doi: 10.1016/0076-6879(75)43136-6. [DOI] [PubMed] [Google Scholar]
  19. Schumacher G., Sizmann D., Haug H., Buckel P., Böck A. Penicillin acylase from E. coli: unique gene-protein relation. Nucleic Acids Res. 1986 Jul 25;14(14):5713–5727. doi: 10.1093/nar/14.14.5713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Vandamme E. J., Voets J. P. Microbial penicillin acylases. Adv Appl Microbiol. 1974;17(0):311–369. doi: 10.1016/s0065-2164(08)70563-x. [DOI] [PubMed] [Google Scholar]
  21. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

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