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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| Published in | FEMS microbiology letters Vol. 206; no. 2; pp. 157 - 162 |
|---|---|
| Main Authors | , , |
| Format | Journal Article |
| Language | English |
| Published |
Oxford, UK
Blackwell Publishing Ltd
01.01.2002
Oxford University Press |
| Subjects | |
| Online Access | Get full text |
| ISSN | 0378-1097 1574-6968 |
| DOI | 10.1111/j.1574-6968.2002.tb11002.x |
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| Abstract | 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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| AbstractList | The dyatic symmetric element (DSE) present in the alpha-amylase gene promoter region of the thermophilic Streptomyces strain sp. TO1, and the whole alpha-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 alpha-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.The dyatic symmetric element (DSE) present in the alpha-amylase gene promoter region of the thermophilic Streptomyces strain sp. TO1, and the whole alpha-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 alpha-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. 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. 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. |
| Author | Bejar, Samir Karray-Rebai, Ines Mellouli, Lotfi |
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| DOI | 10.1111/j.1574-6968.2002.tb11002.x |
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| References_xml | – volume: 140 start-page: 1059 year: 1994 end-page: 1067 article-title: Sequences involved in growth‐phase dependent expression and glucose repression of a α‐amylase gene publication-title: Microbiology – volume: 31 start-page: 426 year: 1959 end-page: 428 article-title: Use of dinitrosalicylic acid reagent for determination of reducing sugars publication-title: Anal. Chem. – volume: 129 start-page: 1747 year: 1983 end-page: 1813 article-title: Numerical classification of and related genera publication-title: J. Gen. Microbiol. – volume: 23 start-page: 537 year: 1997 end-page: 549 article-title: Substrate induction and glucose repression of maltose utilization by A3 is controlled by , a member of the family of regulatory genes publication-title: Mol. Microbiol. – volume: 181 start-page: 31 year: 1999 end-page: 39 article-title: Regulation of the expression of TO1 encoding a thermostable α‐amylase from sp TO1, in its original host and in TK24 publication-title: FEMS Microbiol. Lett. – volume: 56 start-page: 100 year: 1992 end-page: 122 article-title: Cyclic AMP in prokaryotes publication-title: Microbiol. Rev. – volume: 18 start-page: 809 year: 1996 end-page: 814 article-title: Characterisation and molecular cloning of thermostable alpha‐amylase from sp TO1 publication-title: Biotechnol. Lett. – volume: 89 start-page: 1885 year: 1992 end-page: 1889 article-title: Direct repeat sequences are implicated in the regulation of two chitinase promoters that are subject to carbon catabolite control publication-title: Proc. Natl. Acad. Sci. USA – volume: 166 start-page: 557 year: 1983 end-page: 580 article-title: Studies on transformation of with plasmids publication-title: J. Mol. Biol. – volume: 72 start-page: 248 year: 1976 end-page: 254 article-title: A rapid and sensitive method for the quantitation of microgram quantities of protein utilising the principle of protein‐dyebinding publication-title: Anal. Biochem. – volume: 179 start-page: 6383 year: 1997 end-page: 6390 article-title: Amylase and chitinase genes in are regulated by , a pleitropic regulatory gene publication-title: J. Bacteriol. – volume: 104 start-page: 258 year: 1967 end-page: 262 article-title: Isolation of deoxyribonucleic acid and ribosomal ribonucleic acid from bacteria publication-title: J. Biochem. – volume: 28 start-page: 865 year: 1998 end-page: 874 article-title: PRD – a protein domain involved in PTS‐dependent induction and carbon catabolite repression of catabolic operons in bacteria publication-title: Mol. Microbiol. |
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The dyatic symmetric element (DSE) present in the α-amylase gene promoter region of the thermophilic Streptomyces strain sp. TO1, and the whole... 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... 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... The dyatic symmetric element (DSE) present in the alpha-amylase gene promoter region of the thermophilic Streptomyces strain sp. TO1, and the whole... |
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| SubjectTerms | Affinity Amylases Catabolite repression Cloning Cloning vectors Copy number Gene expression Glycerol High copy number LacI‐like regulator Microbiology Plasmids Transcription α-Amylase |
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| Title | Construction of α-amylase-producing strains not subject to carbon catabolite repression |
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