Overproduction of bacterial amylases in recombinant Escherichia coli systems

Seo, Jin‐Ho; Lee, Woo‐Jong; Kim, Myoung-Dong; Kim, Chang-Sup; Park, Yong-Chul

doi:10.1016/s0921-0423(96)80370-4

Cited by 1 publication

(1 citation statement)

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“…A wide variety of organisms including plants, fungi and bacteria are able to represent this enzymatic activity (Janecek 1997). Due to industrial importance, extended types of enzymes from the a-amylase family were subjected to overproduction in different recombinant systems (Sibakov 1986;Suominen et al 1995;Marco et al 1996;Seo et al 1996;Jogensen et al 1997;Lin and Hsu 1997;Park et al 1997). Bacillus licheniformis a-amylase (BLA) is among the most extensively thermostable natural enzymes used in starch technology (Aehle and Misset 1999).…”

Section: Introductionmentioning

confidence: 99%

Expression and secretion of an alpha-amylase gene from a native strain of Bacillus licheniformis in Escherichia coli by T7 promoter and putative signal peptide of the gene

Shahhoseini¹,

Ziaee²,

Ghaemi

2003

J Appl Microbiol

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The gene encoding a hyperthermostable a-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 a-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.

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