Development of a selection system for the detection of L-ribose isomerase expressing mutants of Escherichia coli.

Development of a selection system for the detection of L-ribose isomerase expressing mutants of Escherichia coli. Research Abstract Details 

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Development of a selection system for the detection of L-ribose isomerase expressing mutants of Escherichia coli. Abstract Text:

Cassandra De Muynck,Jef Van der Borght,Marjan De Mey,Sofie L De Maeseneire,Inge N A Van Bogaert,Joeri Beauprez,Wim Soetaert,Erick Vandamme,

L-Arabinose isomerase (E.C. 5.3.1.14) catalyzes the reversible isomerization between L-arabinose and L-ribulose and is highly selective towards L-arabinose. By using a directed evolution approach, enzyme variants with altered substrate specificity were created and screened in this research. More specifically, the screening was directed towards the identification of isomerase mutants with L-ribose isomerizing activity. Random mutagenesis was performed on the Escherichia coli L-arabinose isomerase gene (araA) by error-prone polymerase chain reaction to construct a mutant library. To enable screening of this library, a selection host was first constructed in which the mutant genes were transformed. In this selection host, the genes encoding for L-ribulokinase and L-ribulose-5-phosphate-4-epimerase were brought to constitutive expression and the gene encoding for the native L-arabinose isomerase was knocked out. L-Ribulokinase and L-ribulose-5-phosphate-4-epimerase are necessary to ensure the channeling of the formed product, L-ribulose, to the pentose phosphate pathway. Hence, the mutant clones could be screened on a minimal medium with L-ribose as the sole carbon source. Through the screening, two first-generation mutants were isolated, which expressed a small amount of L-ribose isomerase activity.

Development of a selection system for the detection of L-ribose isomerase expressing mutants of Escherichia coli. Publishing Authors By Initials

C De Muynck,J Van der Borght,M De Mey,SL De Maeseneire,IN Van Bogaert,J Beauprez,W Soetaert,E Vandamme,

For similar genetic processes: selection (genetics) research abstracts see: genetic processes: selection (genetics) research

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Development of a selection system for the detection of L-ribose isomerase expressing mutants of Escherichia coli. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Applied microbiology and biotechnology

VOLUME: 76

Page Numbers: 1051-7

Journal Abbreviation: Appl. Microbiol. Biotechnol.

ISSN: 0175-7598

DAY: 10

MONTH: 07

YEAR: 2007

Development of a selection system for the detection of L-ribose isomerase expressing mutants of Escherichia coli. Information

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LANGUAGE: eng

NlmUniqueID: 8406612

Development of a selection system for the detection of L-ribose isomerase expressing mutants of Escherichia coli. Keywords Mesh Terms:

KEYWORDS: Selection (Genetics)

MESH TERMS: growth & development

Chemical & Substance for Abstract: Development of a selection system for the detection of L-ribose isomerase expressing mutants of Escherichia coli. Information

Substance Name: ribose isomerase

Registry Number: EC 5.3.1.20

Grant and Affiliation Information for Development of a selection system for the detection of L-ribose isomerase expressing mutants of Escherichia coli.

AFFILIATION: Pharmadvize, Kleimoer 4, B-9030, Mariakerke, Belgium. cdmuynck@hotmail.com

Country: Germany

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MEDLINETA: Appl Microbiol Biotechnol

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