Publication: Cloning and expression of penicillin acylase genes from overproducing strains of Escherichia coli and Bacillus megaterium
Issued Date
1987-01-01
Resource Type
ISSN
14320614
01757598
01757598
Other identifier(s)
2-s2.0-0023149799
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Mahidol University
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SCOPUS
Bibliographic Citation
Applied Microbiology and Biotechnology. Vol.25, No.4 (1987), 372-378
Suggested Citation
V. Meevootisom, J. R. Saunders Cloning and expression of penicillin acylase genes from overproducing strains of Escherichia coli and Bacillus megaterium. Applied Microbiology and Biotechnology. Vol.25, No.4 (1987), 372-378. doi:10.1007/BF00252550 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/15325
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Title
Cloning and expression of penicillin acylase genes from overproducing strains of Escherichia coli and Bacillus megaterium
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Abstract
Penicillin acylase genes from Escherichia coli 194, of an overproducing mutant (194-3) of this strain, and of a similar overproducing mutant of Bacillus megaterium UN1 were cloned in E. coli DH1 on the plasmid vector pACYC184. The sizes of chromosomal DNA fragments essential for penicillin acylase production were found by Tn 1000 mutagenesis and in vitro deletions to be between 2.2 and 2.5 kb in the case of both E. coli genes and between 2.3 and 2.7 kb in the case of the mutant Bacillus gene. Restriction mapping indicated substantial sequence differences between the E. coli and B. megaterium penicillin acylase genes. Enzyme production in E. coli recombinants from both overproducing mutants was found to be constitutive and higher than in the original strains. The Bacillus penicillin acylase was produced intracellularly in E. coli recombinants, which is in contrast to the normal extracellular production of this enzyme in B. megaterium. Recombinant plasmids containing penicillin acylase genes from either source were found to be unstable in the absence of selection pressure for retention of the vector. © 1987 Springer-Verlag.