Toyobo Co., Ltd.

Toyo Boseki Kabushiki Kaisha

National Research Institute of Brewing

Takahide Kishimoto

Hiroshi Aiba

Shusaku Yanagidani

Yuu Utashima

Yosuke Sumida

Yoshiaki Nishiya

Hiroshi Kawaminami

Rie Hirao

Masao Kitabayashi

Shizuo Hattori

Masanori Oka

Yuji Tsuji

Yasuhiro Yamazaki

Atsushi Sogabe

Shigenori Emi

Yukihiro Sogabe

Seiji Takeshima

Hiroaki Kitazawa

Yoshihisa Kawamura

Noriaki Hirayama

Sachio Iwai

Tadanobu Matsumura

Atsushi Kawai

Kazuo Masaki

The invention provides streptolysin O containing a polypeptide comprising an amino acid sequence having deletion of a polypeptide extending from a lysine residue at position 2 of SEQ ID NO: 1 to any of an alanine residue at position 31, a glutamate residue at position 32, a serine residue at position 33, and an asparagine residue at position 34, and also having deletion of any of an isoleucine residue at position 465 to an alanine residue at position 472 of SEQ ID NO: 1 and all of the subsequent amino acid residues, as well as variants thereof and related DNA, vectors, transformants, and methods of use thereof.

Modified streptolysin O

A glucose monitoring method and a glucose sensor, both of which use glucose dehydrogenase having a Michaelis constant (Km) for xylose of 600 mM or more and 3000 mM or less, and a Km for glucose of 0.1 mM or more and 100 mM or less, which provide for evaluating FADGDH in an aqueous system while reducing the practical influence of FADGDH on D-xylose.

Glucose monitoring method and glucose sensor

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

Object An object of the present invention is to provide a highly stable FVHO preparation and a low-hygroscopicity dried FVHO preparation. Means for achieving the object A method for producing a FVHO preparation comprising a step of allowing at least one member selected from phosphoric acid, casein peptone, D-glucosamine hydrochloride, melibiose, sorbose, lactose, fructose, melezitose, glucono-1,5-lactone, and ribitol; and a method for producing a dried FVHO preparation, comprising a step of allowing Bicine to coexist.

Method for producing fructosyl valyl histidine oxidase preparation

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.

A novel diaphorase; a method for producing the diaphorase; and use of the diaphorase are provided. The diaphorase comprises any one of the following polypeptides (a) to (c): (a) a polypeptide having the amino acid sequence of SEQ ID NO: 1, (b) a polypeptide having the amino acid sequence of SEQ ID NO: 1 in which one or several amino acid residues are substituted, deleted, inserted, added, and/or inverted, and having diaphorase activity, and (c) a polypeptide having an amino acid sequence with 80% or more identity to the amino acid sequence of SEQ ID NO: 1, and having diaphorase activity.

Diaphorase

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 AP that has a high specific activity and preferably has superior reactivity with respect to various luminescent substrates generally used for high-sensitivity immunoassay analysis. Further preferably, the present invention provides AP having a thermal stability higher than that of CIAP. The alkaline phosphatase of the present invention is derived from the genusand has the following characteristics: (A) molecular weight: about 104,000; (B) optimum reaction pH: about 9.5; (C) stable pH range: 5.5 to 10.4; (D) thermal stability: 65&#xb0; C., and (E) specific activity: 5,000 U/mg or more.

Alkaline phosphatase

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.

Basidiomycetous yeast mutant

The present invention provides glucose dehydrogenase which is excellent in heat resistance and substrate specificity and is not affected by dissolved oxygen. Specifically, the present invention relates to glucose dehydrogenase characterized by being derived from an eukaryotic organism and keeping 90% or more activity after being treated with heat at 55&#xb0; C. for 15 minutes in a liquid form compared with the activity before being treated.

The present invention provides a method for highly expressing a recombinant FAD-GDH protein derived from filamentous fungi, protein obtained by the method, and a regent for measuring glucose using the protein. According to the invention, the FAD-GDH can be highly expressed by altering DNA sequence coding for a signal peptide of FAD-GDH gene isolated from. FAD-GDH can be stably produced by adjusting pH of 7.1 to 7.3 during culture production.

Growing community of inventors

Tsuruga, Japan

Takahide Kishimoto