Cloning, characterisation, and heterologous expression of the Candida utilis malic enzyme gene

Date
2006
Authors
Saayman M.
van Zyl W.H.
Viljoen-Bloom M.
Journal Title
Journal ISSN
Volume Title
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Abstract
The Candida utilis malic enzyme gene, CME1, was isolated from a cDNA library and characterised on a molecular and biochemical level. Sequence analysis revealed an open reading frame of 1,926 bp, encoding a 641 amino acid polypeptide with a predicted molecular weight of approximately 70.2 kDa. The inferred amino acid sequence suggested a cytosolic localisation for the malic enzyme, as well as 37 and 68% homologies with the malic enzymes of Schizosaccharomyces pombe and Saccharomyces cerevisiae, respectively. Expression of the CME1 gene was subject to carbon catabolite repression and substrate induction, similar to the regulatory mechanisms observed for the C. utilis dicarboxylic acid permease. The CME1 gene was successfully expressed in S. cerevisiae under control of the S. cerevisiae PGK1 promoter and terminator. When coexpressed with the S. pombe malate permease gene (mae1), it resulted in a recombinant S. cerevisiae strain able to completely degrade 90% of the extracellular L-malate within 24 h. © Springer-Verlag 2006.
Description
Keywords
amino acid, carbon, complementary DNA, dicarboxylic acid, malate dehydrogenase (decarboxylating), malic acid, permease, polypeptide, fungal protein, malate dehydrogenase, recombinant protein, amino acid sequence, article, Candida utilis, catabolite repression, chemical analysis, controlled study, cytosol, DNA library, enzyme induction, enzyme isolation, enzyme localization, enzyme substrate, extracellular space, fungal strain, genetic analysis, genetic code, genetic regulation, heterologous expression, molecular cloning, molecular weight, nonhuman, nucleotide sequence, open reading frame, prediction, priority journal, promoter region, regulatory mechanism, Saccharomyces cerevisiae, Schizosaccharomyces pombe, sequence analysis, sequence homology, biosynthesis, Candida, enzymology, gene expression, genetics, molecular genetics, Schizosaccharomyces, Pichia jadinii, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Amino Acid Sequence, Candida, Cloning, Molecular, Fungal Proteins, Gene Expression, Malate Dehydrogenase, Molecular Sequence Data, Recombinant Proteins, Saccharomyces cerevisiae, Schizosaccharomyces, Sequence Homology, Amino Acid
Citation
Current Genetics
49
4