Construction of α-amylase-producing strains not subject to carbon catabolite repression

Abstract The dyatic symmetric element (DSE) present in the α-amylase gene promoter region of the thermophilic Streptomyces strain sp. TO1, and the whole α-amylase gene (amy TO1) with a 3-bp change in the DSE, were cloned in the high copy number replicative cloning vector pIJ702 giving pLM10 and pLM1...

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Bibliographic Details
Published inFEMS microbiology letters Vol. 206; no. 2; pp. 157 - 162
Main Authors Mellouli, Lotfi, Karray-Rebai, Ines, Bejar, Samir
Format Journal Article
LanguageEnglish
Published Oxford, UK Blackwell Publishing Ltd 01.01.2002
Oxford University Press
Subjects
Affinity
Amylases
Catabolite repression
Cloning
Cloning vectors
Copy number
Gene expression
Glycerol
High copy number
LacI‐like regulator
Microbiology
Plasmids
Transcription
α-Amylase
Affinity
α-Amylase
Gene expression
LacI-like regulator
High copy number
Online AccessGet full text
ISSN0378-1097
1574-6968
DOI10.1111/j.1574-6968.2002.tb11002.x

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Summary:Abstract The dyatic symmetric element (DSE) present in the α-amylase gene promoter region of the thermophilic Streptomyces strain sp. TO1, and the whole α-amylase gene (amy TO1) with a 3-bp change in the DSE, were cloned in the high copy number replicative cloning vector pIJ702 giving pLM10 and pLM11 plasmids respectively. In TO1/pLM10 and Streptomyces lividans TK24/pLM11 strains, the expression of α-amylase TO1 gene became insensible to the negative effect of glucose and glycerol. These results strongly suggest that, in a high copy number system, the negative transcriptional regulator was titrated out by the DSE and the repression of the expression of amy TO1 gene is abolished.
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ISSN:0378-1097
1574-6968
DOI:10.1111/j.1574-6968.2002.tb11002.x