Characterization of two aminotransferases from Candida albicans

• Authors: Kamila Rząd , Iwona Gabriel
• Languages of publication: EN

Abstracts

EN

Aminoadipate aminotransferase (AmAA) is an enzyme of α-aminoadipate pathway (AAP) for l-lysine biosynthesis. AmAA may also participated in biosynthesis or degradation of aromatic amino acids and in d-tryptophan based pigment production. The AAP is unique for fungal microorganisms. Enzymes involved in this pathway have specific structures and properties. These features can be used as potential molecular markers. Enzymes catalyzing reactions of l-lysine biosynthesis in Candida albicans may also become new targets for antifungal chemotherapy. Search of the NCBI database resulted in identification of two putative aminoadipate aminotransferase genes from Candida albicans: ARO8 (ORFs 19.2098 and 19.9645) and YER152C (ORFs 19.1180 and 19.8771). ARO8 from C. albicans exhibits 53% identity to ARO8 from S. cerevisiae, while YER152C exhibits 30% identity to ARO8 and 45% to YER152C from S. cerevisiae. We amplified two genes from the C. albicans genome: ARO8 and YER152C. Both were cloned and expressed as His-tagged fusion proteins in E. coli. The purified Aro8CHp gene product revealed aromatic and α-aminoadipate aminotransferase activity. Basic molecular properties of the purified protein were determined. We obtained catalytic parameters of Aro8CHp with aromatic amino acids and aminoadipate (AA) (Km(L-Phe) 0.05±0.003 mM, Km(L-Tyr) 0.1±0.008 mM, Km(L-AA) 0.02±0.006 mM) and confirmed the enzyme broad substrate spectrum. The assays also demonstrated that this enzyme may use 2-oxoadipate and 2-oxoglutarate (2-OG) as amino acceptors. Aro8-CHp exhibited pH optima range of 8, which is similar to AmAA from S. cerevisiae. Our results also indicate that CaYer152Cp has a possible role only in aromatic amino acids degradation, in contrast to CaAro8CHp.

Keywords

EN

l-lysine biosynthesis; aminoadipate aminotransferase; aromatic aminotransferase; Candida albicans

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2015
• Volume: 62
• Issue: 4
• Pages: 903-912

Dates

published

2015

received

2015-07-31

revised

2015-10-09

accepted

2015-10-29

(unknown)

2015-11-30

Contributors

author

Kamila Rząd

• Department of Pharmaceutical Technology and Biochemistry, Gdansk University of Technology, Gdańsk, Poland

author

Iwona Gabriel

• Department of Pharmaceutical Technology and Biochemistry, Gdansk University of Technology, Gdańsk, Poland

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Identifiers

DOI

10.18388/abp.2015_1158