Open AccessNucleotide sequence of the gene encoding the Streptomyces albus G β‐lactamase precursor
Author(s) -
DEHOTTAY Philippe,
DUSART Jean,
MEESTER Fabien,
JORIS Bernard,
BEEUMEN Jozef,
ERPICUM Thomas,
FRÈRE JeanMarie,
GHUYSEN JeanMarie
Publication year - 1987
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1987.tb13521.x
Subject(s) - streptomyces albus , biology , serine , nucleic acid sequence , ribosomal binding site , biochemistry , peptide sequence , microbiology and biotechnology , gene , amino acid , structural gene , streptomyces , mutant , ribosome , genetics , enzyme , bacteria , rna
A 1400‐base DNA fragment, which contains the gene encoding the extracellular active‐site serine β‐lactamase of Streptomyces albus G previously cloned into Streptomyces lividans [Dehottay et al. (1986) Gene 42 , 31–36], was sequenced. The gene codes for a 314‐amino‐acid precursor, the N‐terminal region of which has the characteristics of a signal peptide. The β‐lactamase as excreted by the host strain S. lividans PD6 has a ragged N‐terminus, indicating either the presence of a leader peptidase of poor specificity or the action of an aminopeptidase. The primary structure (as deduced from the nucleotide sequence) was confirmed by amino acid sequencing of a 16‐residue stretch at the amino terminus of the protein, a 12‐residue stretch containing the active‐site serine [De Meester et al. (1987) Biochem. J. 244 , 427–432] and a 23‐residue stretch obtained by trypsin digestion of the protein. The β‐lactamase belongs to class A, has three half‐cystine residues (one of which occurs on the amino side of the active‐site serine) and is inactivated by thiol reagents. Pultative ribosome binding site and terminator region were identified.