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KMID : 0545120040140030547
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Journal of Microbiology and Biotechnology
2004 Volume.14 No. 3 p.547 ~ p.552
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Engineering Recombinant Streptomyces coelicolor Malate Synthase with Improved Thermal Properties by Directed Mutagenesis
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Koh R
Goh LL/Sim TS
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Abstract
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Streptomyces thermovulgaris malate synthase (stMS) is known to be more thermostable and thermoactive than S. coelicolor malate synthase (scMS). Therefore based on the amino acid sequence of stMS 3 scMS mutants namely P186R T8PL9P and T8PL9PP186R were created by site-directed mutagenesis in an attempt to engineer a more thermoactive and thermostable enzyme. An enzymatic analysis of the wild-type and mutant MS revealed that P186R and T8PL9PP186R were more thermoactive than the wild-type scMS and T8PL9P. Furthermore all 3 mutants exhibited a greater thermostability than scMS thereby suggesting that both R186 and P8P9 can cause increased thermostability in scMS.
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KEYWORD
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