Novel glucose dehydrogenase from Mucor prainii: Purification, characterization, molecular cloning and gene expression in Aspergillus sojae.
Glucose dehydrogenase (GDH) is of interest for its potential applications in the field of glucose sensors. To improve the performance of glucose sensors, GDH is required to have strict substrate specificity. A novel flavin adenine dinucleotide (FAD)-dependent GDH was isolated from Mucor prainii NISL0103 and its enzymatic properties were characterized. This FAD-dependent GDH (MpGDH) exhibited high specificity toward glucose. High specificity for glucose was also observed even in the presence of saccharides such as maltose, galactose and xylose. The molecular masses of the glycoforms of GDH ranged from 90 to 130 kDa. After deglycosylation, a single 80 kDa band was observed. The gene encoding MpGDH was cloned and expressed in Aspergillus sojae. The apparent kcat and Km values of recombinant enzyme for glucose were found to be 749.7 s(-1) and 28.3 mM, respectively. The results indicated that the characteristics of MpGDH were suitable for assaying blood glucose levels.
Research and Development Division, Kikkoman Corporation, 399 Noda, Chiba 278-0037, Japan. Electronic address: rsatake@mail.kikkoman.co.jp.
Ryoko
R
Satake
Research and Development Division, Kikkoman Corporation, 399 Noda, Chiba 278-0037, Japan.
Atsushi
A
Ichiyanagi
Research and Development Division, Kikkoman Corporation, 399 Noda, Chiba 278-0037, Japan.
Keiichi
K
Ichikawa
Research and Development Division, Kikkoman Corporation, 399 Noda, Chiba 278-0037, Japan.
Kozo
K
Hirokawa
Research and Development Division, Kikkoman Corporation, 399 Noda, Chiba 278-0037, Japan.
Yasuko
Y
Araki
Research and Development Division, Kikkoman Corporation, 399 Noda, Chiba 278-0037, Japan.
Taro
T
Yoshimura
Research and Development Division, Kikkoman Corporation, 399 Noda, Chiba 278-0037, Japan.
Keiko
K
Gomi
D001230:Aspergillus / Q000235:genetics*; D003001:Cloning, Molecular; D005182:Flavin-Adenine Dinucleotide / Q000378:metabolism; D005690:Galactose / Q000378:metabolism / Q000494:pharmacology; D015870:Gene Expression; D005947:Glucose / Q000378:metabolism* / Q000494:pharmacology; D042843:Glucose 1-Dehydrogenase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D006031:Glycosylation; D008320:Maltose / Q000378:metabolism / Q000494:pharmacology; D008970:Molecular Weight; D009089:Mucor / Q000201:enzymology*; D013379:Substrate Specificity / Q000187:drug effects; D014994:Xylose / Q000378:metabolism / Q000494:pharmacology
D005182:Flavin-Adenine Dinucleotide; D008320:Maltose; D014994:Xylose; D042843:Glucose 1-Dehydrogenase; D005947:Glucose; D005690:Galactose
Diabetes; Flavin adenine dinucleotide; Glucose dehydrogenase; Glucose sensor; Substrate specificity