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Title: PCR-mediated screening and cloning of a malate synthase gene from Streptomyces griseus NCIMB 9001
Authors: Loke, P. 
Wee, J.
Keng Ing Seah
Sim, T.-S. 
Keywords: Acetate
Malate synthase
PCR cloning
Streptomyces griseus
Issue Date: 2001
Citation: Loke, P., Wee, J., Keng Ing Seah, Sim, T.-S. (2001). PCR-mediated screening and cloning of a malate synthase gene from Streptomyces griseus NCIMB 9001. World Journal of Microbiology and Biotechnology 17 (6) : 645-649. ScholarBank@NUS Repository.
Abstract: Malate synthase is an essential metabolic enzyme of the glyoxylate bypass that makes possible the replenishment of carbon intermediates to cells grown on acetate. A polymerase chain reaction (PCR)-based molecular screening investigation of full-length malate synthase genes from Streptomyces spp. was initiated by our group. To this end, consensus primers were designed based on known streptomycete malate synthase sequences and successful amplification was obtained for Streptomyces griseus, S. fimbriatus and S. lipmanii. The putative full-length malate synthase gene from S. griseus was subsequently cloned, sequenced and expressed. Sequence analysis of this gene showed very high identity with other streptomycete malate synthase genes. Furthermore, high malate synthase activity was detected after heterologous expression in Escherichia coli, thus demonstrating successfully the rapid cloning and functional verification of a streptomycete malate synthase gene. Growth studies of S. griseus revealed that malate synthase activity was induced by the presence of acetate, which is a two-carbon source. Interestingly, the activity peaked during late growth phase when the biomass was declining, suggesting that the enzyme may have a late role in metabolism.
Source Title: World Journal of Microbiology and Biotechnology
ISSN: 09593993
DOI: 10.1023/A:1012419705929
Appears in Collections:Staff Publications

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