Toyobo Co., Ltd.

National Research Institute of Brewing

Yuu Utashima

Takahide Kishimoto

Hiroshi Aiba

Shusaku Yanagidani

Yosuke Sumida

Rie Hirao

Hiroshi Kawaminami

Yasuhiro Yamazaki

Hiroaki Kitazawa

Kazuo Masaki

An object of the present invention is to construct a more excellent glucose sensor, and to provide GDH more suitable for the glucose sensor. Provided is FAD-dependent glucose dehydrogenase in which the range of molecular weight distribution observed by SDS-PAGE is within 50 kDa when viewed in a molecular weight distribution in which the relative value of band intensity exceeds 60% of the maximum value.

FAD-dependent glucose dehydrogenase

Provided is GDH with increased applicability to glucose sensors. A composition contains FADGDH, wherein when 0.1 mL of the composition is added to 2.9 mL of a solution containing 10 mM of trehalose and 1 mmol/L of potassium ferricyanide to give a glucose dehydrogenase activity of 500 U/mL and incubated at 37&#xb0; C., the decrease in absorbance at 405 nm resulting from reduction of the potassium ferricyanide is less than 20 mAbs per minute.

Enzyme preparation for use in measurement of glucose

The present invention provides a novel glucose dehydrogenase that has excellent substrate specificity and that is suitable for use in SMBG. The present invention provides a flavin-bound glucose dehydrogenase having the following characteristics (1) to (5): (1) Temperature stability: stable at a temperature of 45&#xb0; C. or lower; (2) stable at a pH range of 4.5 to 7.5; (3) substrate specificity: the reactivity to D-xylose, maltose, or D-galactose is 2% or less, based on the reactivity to D-glucose taken as 100%; (4) optimal activity temperature: 34 to 47&#xb0; C.; and (5) optimal activity pH: 6.3 to 6.7.

Glucose dehydrogenase

Provided is a flavin adenine dinucleotide-dependent glucose dehydrogenase comprising a polypeptide having an amino acid sequence with 78% or more identity to the amino acid sequence of SEQ ID NO: 3, and having glucose dehydrogenase activity.

The present invention provides a novel glucose dehydrogenase that has excellent substrate specificity, specific activity, thermal stability, and the like, and that is suitable for use in SMBG sensors. The present invention provides a purified polypeptide comprising an amino acid sequence having at least 80% identity to the sequence of SEQ ID NO: 1 and having glucose dehydrogenase activity.

An object of the present invention is to provide a novel glucose dehydrogenase, a method for producing the glucose dehydrogenase, and applications of the glucose dehydrogenase. An isolated flavin-binding glucose dehydrogenase comprising a polypeptide having an amino acid sequence with 80% or more identity to the amino acid sequence of SEQ ID NO: 1, and having glucose dehydrogenase activity is provided.

An object of the present invention is to provide a novel glucose dehydrogenase, a method for producing the glucose dehydrogenase, and applications of the glucose dehydrogenase. The flavin-binding glucose dehydrogenase of the invention has the following characteristics (1) and (4): (1) Molecular weight: the molecular weight of a polypeptide moiety in the enzyme is about 68 kDa as measured by SDS-polyacrylamide electrophoresis; (2) Km value: the Km value for D-glucose is about 15 mM or less; (3) Temperature stability: stable at a temperature of 55&#xb0; C. or less; and (4) pH stability: stable at a pH range of 3.0 to 8.5.

The present invention provides a host for genetic recombination useful in the production of heterologous proteins in a large scale, by suppressing the production of extracellular polysaccharides in the basidiomycetous yeasts.sp. S-2 D11 strain (FERM BP-11482) which is characterized in that the production of extracellular polysaccharides is suppressed as compared with the parent strain.

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Tsuruga, Japan

Yuu Utashima