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Expression and secretion of an α‐amylase gene from a native strain of Bacillus licheniformis in Escherichia coli by T7 promoter and putative signal peptide of the gene
Author(s) -
Shahhoseini M.,
Ziaee A.A.,
Ghaemi N.
Publication year - 2003
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1046/j.1365-2672.2003.02082.x
Subject(s) - bacillus licheniformis , signal peptide , escherichia coli , biology , signal peptidase , microbiology and biotechnology , biochemistry , extracellular , recombinant dna , peptide , amylase , alpha amylase , expression vector , gene , enzyme , bacillus subtilis , bacteria , genetics
The gene encoding a hyperthermostable α ‐amylase from a Bacillus licheniformis native strain was cloned in pET24d transcription vector containing T7 promoter, and expressed in Escherichia coli BL21(DE3) cells. Having confirmed the α ‐amylase activity through activity staining method on SDS–PAGE gel, the yields of production were determined in two separated intra and inter‐cellular phases and compared using enzymatic assay methods. Extracellular production of the active recombinant enzyme implies the recognition of the putative signal peptide of this Bacillus sp. by E. coli secretory system. This may be because of the amino acid sequence of this signal peptide which covers all the structural parameters of a standard signal peptide processed by Lep B, the major signal peptidase in E. coli secretory system. This study recommends the use of this signal peptide for extracellular production of other foreign proteins in E. coli .