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Cloning and Functional Characterization of the Cis-Aconitic Acid Decarboxylase (CAD) Gene From Aspergillus Terreus

Cloning and Functional Characterization of the Cis-Aconitic Acid Decarboxylase (CAD) Gene From Aspergillus Terreus

Shin Kanamasa, Lies Dwiarti, Mitsuyasu Okabe, Enoch Y Park

Appl Microbiol Biotechnol. 2008 Aug;80(2):223-9.

PMID: 18584171

Abstract:

A filamentous fungus Aspergillus terreus produces itaconic acid, which is predicted to be derived from cis-aconitic acid via catalysis by cis-aconitic acid decarboxylase (CAD) in the carbon metabolism of the fungus. To clarify the enzyme's function and a pathway for itaconic acid biosynthesis, we cloned a novel gene encoding the enzyme. The open reading frame of this gene (CAD1) consists of 1,529 bp encoding 490 amino acids and is interrupted by a single intron. Among the identified proteins in the database, the primary structure of the protein encoded by CAD1 shared high identity with the MmgE/PrpD family of proteins, including a number of 2-methylcitrate dehydratases of bacteria. The cloned gene excluding an intron was introduced into the expression plasmid pAUR-CAD1 controlled by the ADH1 promoter. The CAD activity in Saccharomyces cerevisiae was confirmed by directly detecting itaconic acid as a product from cis-aconitic acid as a substrate. This result reveals for the first time that this gene encodes CAD, which is essential for itaconic acid production in A. terreus.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP585842	cis-Aconitic acid		585-84-2	Price

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