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PMID: 3124817 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

beta-lactamase I from Bacillus cereus. Structure and site-directed mutagenesis.

The Biochemical journal ·Vol. 248 ·No. 3 ·1987-12-15 ·Pages 657-62

Madgwick PJ, Waley SG

Abstract

The sequence of the gene for beta-lactamase I from Bacillus cereus 569/H has been redetermined. Oligonucleotide-directed mutagenesis has been carried out, and the effects of the changes on the ampicillin-resistance of Escherichia coli TG1 expressing the mutant genes have been studied. Lysine-73, close to the active-site serine-70 and a highly-conserved residue, has been converted into arginine. This change had a large effect on activity, but did not abolish it. An even larger effect was found in the mutant in which glutamate-166 had been converted into glutamine; this had little or no activity. On the other hand, the conversion of glutamate-168 into aspartate gave fully active enzyme. Glutamate-166 is an invariant residue, but glutamate-168 is not. Alanine-123 has been replaced by cysteine, to give active enzyme; this change forms part of the plan to introduce a disulphide bond into the enzyme.

MeSH Terms
Ampicillin Resistance/genetics Bacillus cereus/enzymology,genetics Base Sequence Binding Sites Escherichia coli/genetics Molecular Sequence Data Mutation Oligosaccharides/biosynthesis Penicillinase/genetics
Chemicals
Oligosaccharides Penicillinase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Madgwick P J
Sir William Dunn School of Pathology, University of Oxford, U.K.
Waley S G
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2480657
Published
1987-12-15
Pages
657-62
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1148599
Subset
IM
Databases
GENBANK
X06599
External Links
PubMed source DOI: 10.1042/bj2480657 PMC Full Text View journal details
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