Physiological role of the D-amino acid oxidase gene, DAO1, in carbon and nitrogen metabolism in the methylotrophic yeast Candida boidinii
✍ Scribed by Hiroya Yurimoto; Tetsuya Hasegawa; Yasuyoshi Sakai; Nobuo Kato
- Publisher
- John Wiley and Sons
- Year
- 2000
- Tongue
- English
- Weight
- 376 KB
- Volume
- 16
- Category
- Article
- ISSN
- 0749-503X
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✦ Synopsis
A methylotrophic yeast, Candida boidinii, exhibits D-amino acid oxidase activity (DAO, EC 1.4.3.3) during its growth on D-alanine as a sole carbon or a nitrogen source. The structural gene (DAO1), encoding DAO, was cloned from a genomic library of C. boidinii.
The 1035-bp gene encoded 345 amino acids and the predicted amino acid sequence showed signi®cant similarity to those of DAOs from other organisms. The DAO1 gene was disrupted in the C. boidinii genome by one-step gene disruption. The DAO1-deleted strain did not grow on D-alanine as a carbon source but did grow on D-alanine as a sole nitrogen source (with glucose as the carbon source). These results suggested that, while DAO is critically involved in growth on D-alanine as a carbon source, there should be another enzyme system which metabolizes D-alanine as a nitrogen source in C. boidinii. We also showed that the three C-terminal amino acid sequence of DAO, xAKL was necessary and suf®cient for the import of DAO into peroxisomes.