Activation of a peroxisomal Pichia pastoris d-amino acid oxidase, which uses d-alanine as a preferred substrate, depends on pyruvate carboxylase

Sandra H. Klompmaker, Aysun Kilic, Richard J. Baerends, Marten Veenhuis, Ida J. van der Klei*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

d-Amino acid oxidase (DAO) is an important flavo-enzyme that catalyzes the oxidative deamination of d-amino acids into the corresponding alpha-keto acid, ammonia and H(2)O(2). We identified two amino acid oxidases in the methylotrophic yeast Pichia pastoris: Dao1p, which preferentially uses d-alanine as a substrate, and Dao2p, which uses d-aspartate as a preferred substrate. Dao1p has a molecular mass of 38.2 kDa and a pH optimum of 9.6. This enzyme was localized to peroxisomes, albeit a typical peroxisomal targeting signal is lacking. Interestingly, P. pastoris mutant strains, defective in the enzyme pyruvate carboxylase, showed a pronounced growth defect on d-alanine, concomitant with a significant reduction in Dao1p activity relative to the wild-type control. This indicates that pyruvate carboxylase functions in import and/or activation of P. pastoris Dao1p.

Original languageEnglish
Pages (from-to)708-716
Number of pages9
JournalFems Yeast Research
Volume10
Issue number6
DOIs
Publication statusPublished - Sep-2010

Keywords

  • d-amino acid oxidase
  • peroxisome
  • pyruvate carboxylase
  • Pichia pastoris
  • moonlighting
  • HANSENULA-POLYMORPHA
  • SACCHAROMYCES-CEREVISIAE
  • TARGETING SIGNAL
  • CANDIDA-BOIDINII
  • ALCOHOL OXIDASE
  • METHANOL OXIDASE
  • YEAST
  • GENE
  • PROTEIN
  • GROWTH

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