Production of fungal fructosyl amino acid oxidase useful for diabetic diagnosis in the peroxisome of Candida boidinii.
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 aod1Delta) 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.
Sakai
Y
Y
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake, Sakyo-ku, Kyoto, Japan. ysakai@kais.kyoto-u.ac.jp
Yoshida
H
H
Yurimoto
H
H
Yoshida
N
N
Fukuya
H
H
Takabe
K
K
Kato
N
N
D000594:Amino Acid Oxidoreductases / Q000096:biosynthesis* / Q000235:genetics; D002175:Candida / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D003920:Diabetes Mellitus / Q000175:diagnosis; D010407:Penicillium / Q000201:enzymology; D020675:Peroxisomes / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis
D011994:Recombinant Proteins; D000594:Amino Acid Oxidoreductases; C103012:amadoriase