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PMID: 3930467 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Cloning and sequencing of the metallothioprotein beta-lactamase II gene of Bacillus cereus 569/H in Escherichia coli.

Journal of bacteriology ·Vol. 164 ·No. 1 ·1985-10-00 ·Pages 223-9

Hussain M, Carlino A, Madonna MJ, Lampen JO

Abstract

The structural gene for beta-lactamase II (EC 3.5.2.6), a metallothioenzyme, from Bacillus cereus 569/H (constitutive for high production of the enzyme) was cloned in Escherichia coli, and the nucleotide sequence was determined. This is the first class B beta-lactamase whose primary structure has been reported. The amino acid sequence of the exoenzyme form, deduced from the DNA, indicates that beta-lactamase II, like other secreted proteins, is synthesized as a precursor with a 30-amino acid N-terminal signal peptide. The pre-beta-lactamase II (Mr, 28,060) is processed in E. coli and in B. cereus to a single mature protein (Mr, 24,932) which is totally secreted by B. cereus but in E. coli remains intracellular, probably in the periplasm. The expression of the gene in E. coli RR1 on the multicopy plasmid pRWHO12 was comparable to that in B. cereus, where it is presumably present as a single copy. The three histidine residues that are involved (along with the sole cysteine of the mature protein) in Zn(II) binding and hence in enzymatic activity against beta-lactams were identified. These findings will help to define the secondary structure, mechanism of action, and evolutionary lineage of B. cereus beta-lactamase II and other class B beta-lactamases.

MeSH Terms
Amino Acid Sequence Bacillus cereus/enzymology,genetics Base Sequence Cephalosporinase/analysis,genetics Cloning, Molecular Escherichia coli/enzymology,genetics Genes, Bacterial beta-Lactamases/genetics
Chemicals
Cephalosporinase beta-Lactamases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Hussain M
Carlino A
Madonna M J
Lampen J O
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42 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.164.1.223-229.1985
Published
1985-10-00
Pages
223-9
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC214233
Subset
IM
Databases
GENBANK
M11189
External Links
PubMed source DOI: 10.1128/jb.164.1.223-229.1985 PMC Full Text View journal details
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