Production of fungal fructosyl amino acid oxidase useful for diabetic diagnosis in the peroxisome of Candida boidinii

FEBS Letters

Kato, Nobuo¹  Sakai, Yasuyoshi¹  Yoshida, Hiroyuki¹  Takabe, Keiji²  Fukuya, Hiroshi¹  Yoshida, Nobuyuki¹  Yurimoto, Hiroya¹ 

[1] Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku, Kyoto 606-8502, Japan;Division of Forest and Biomaterials Science, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku, Kyoto 606-8502, Japan

关键词: Glycation;    Diabetes;    Diagnosis;    Peroxisome;    Methylotrophic yeast;    Gene expression;    FAOD;    fructosyl amino acid oxidase;    FAD;    flavin adenine dinucleotide;    strain aod1Δ;    the alcohol oxidase-depleted strain of the methylotrophic yeast Candida boidinii;    ODC;    orotidylate decarboxylase;    SDS-PAGE;    sodium dodecyl sulfate-polyacrylamide gel electrophoresis;   

DOI  :  10.1016/S0014-5793(99)01245-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A high-level production of fructosyl amino acid oxidase (FAOD), whose production was toxic in Escherichia coli, was investigated through attempts to utilize the peroxisome of Candida boidinii as the place for protein accumulation. The alcohol oxidase-depleted strain (strain aod1Δ) produced FAOD at a four to five times higher level than the wild type strain in terms of protein amount and enzyme activity, although the transcriptional level was similar. As a result of this study, we could improve FAOD productivity approximately 47-fold from the original transformant, and FAOD accumulated within membrane-bound peroxisomes up to 18% of the total soluble proteins.

【 授权许可】

Unknown