The present invention provides members that produce on a large scale a coenzyme-linked glucose dehydrogenase which has excellent substrate-recognizing ability toward glucose while providing low action on maltose. The present invention relates to a polynucleotide encoding a soluble coenzyme-linked glucose dehydrogenase that catalyzes the oxidation of glucose in the presence of an electron acceptor and has an activity toward maltose of 5% or lower; a polypeptide encoded by the nucleotide sequence of the polynucleotide; a recombinant vector carrying the polynucleotide; a transformed cell produced using the recombinant vector; a method for producing a polypeptide comprising culturing the transformed cell and collecting from the cultivated products a polypeptide that links to FAD to exert the glucose dehydration activity; a method for determination of glucose using the polypeptide; a reagent composition for determination of glucose; and a biosensor.

The present invention provides members that produce on a large scale a coenzyme-linked glucose dehydrogenase which has excellent substrate-recognizing ability toward glucose while providing low action on maltose. The present invention relates to a polynucleotide encoding a soluble coenzyme-linked glucose dehydrogenase that catalyzes the oxidation of glucose in the presence of an electron acceptor and has an activity toward maltose of 5% or lower; a polypeptide encoded by the nucleotide sequence of the polynucleotide; a recombinant vector carrying the polynucleotide; a transformed cell produced using the recombinant vector; a method for producing a polypeptide comprising culturing the transformed cell and collecting from the cultivated products a polypeptide that links to FAD to exert the glucose dehydration activity; a method for determination of glucose using the polypeptide; a reagent composition for determination of glucose; and a biosensor.

The present invention provides members that produce on a large scale a coenzyme-linked glucose dehydrogenase which has excellent substrate-recognizing ability toward glucose while providing low action on maltose. The present invention relates to a polynucleotide encoding a soluble coenzyme-linked glucose dehydrogenase that catalyzes the oxidation of glucose in the presence of an electron acceptor and has an activity toward maltose of 5% or lower; a polypeptide encoded by the nucleotide sequence of the polynucleotide; a recombinant vector carrying the polynucleotide; a transformed cell produced using the recombinant vector; a method for producing a polypeptide comprising culturing the transformed cell and collecting from the cultivated products a polypeptide that links to FAD to exert the glucose dehydration activity; a method for determination of glucose using the polypeptide; a reagent composition for determination of glucose; and a biosensor.

Processes, systems and microorganisms are described herein for producing glycolate from ethylene glycol. The processes generally comprise supplying a fermentation broth into a fermentation vessel, wherein the fermentation broth comprises ethylene glycol and a microorganism having a functional metabolic pathway for utilizing ethylene glycol as a carbon source. In a growth phase, an oxygen-containing gas is injected into the fermentation broth to provide oxygen bio-availability conditions to promote cell growth of the microorganism and limit accumulation of glycolate in the fermentation broth. In a production phase, an oxygen-containing gas is injected into the fermentation broth to provide oxygen bio-availability conditions to promote production of glycolate from ethylene glycol by the microorganism and accumulation of the glycolate in the fermentation broth, to produce a glycolate enriched broth.

The present invention provides: a reagent composition having higher storage stability; a sensor involving the reagent composition; and others. According to the present invention, a specific heterocyclic compound is added to a reagent composition to improve the storage stability of the reagent composition and reduce the degree of fluctuation in current values in a sensor that utilizes reagent composition.

The present invention provides members that produce on a large scale a coenzyme-linked glucose dehydrogenase which has excellent substrate-recognizing ability toward glucose while providing low action on maltose. The present invention relates to a polynucleotide encoding a soluble coenzyme-linked glucose dehydrogenase that catalyzes the oxidation of glucose in the presence of an electron acceptor and has an activity toward maltose of 5% or lower; a polypeptide encoded by the nucleotide sequence of the polynucleotide; a recombinant vector carrying the polynucleotide; a transformed cell produced using the recombinant vector; a method for producing a polypeptide comprising culturing the transformed cell and collecting from the cultivated products a polypeptide that links to FAD to exert the glucose dehydration activity; a method for determination of glucose using the polypeptide; a reagent composition for determination of glucose; and a biosensor.

In some embodiments, the present invention provides a protein comprising amino acids in the following sequence L(X).sub.n=14X.sup.3(X).sub.n=1X.sup.1(X).sub.n=1E(X).sub.n=4P(X).sub.n=1NR(X).sub.n=3S(X).sub.n=4D(X).sub.n=2G(X).sub.n=7Y(X).sub.n=4Y (X).sub.n=32-34X.sup.2 (SEQ ID NO: 128), wherein each X independently represents any naturally occurring amino acid residue and n indicates the number of amino acid residues represented by the respective paranthetical at that position, wherein: a) X.sup.1 is selected from the group consisting of S, C, T, M, V, Y, N, P, L, G, Q, A, I, D, W, H, or E, wherein if X.sup.1 is L, H or V, then X.sup.3 is D; and/or b) X.sup.2 is selected from the group consisting of H, L, S or V. In some embodiments, the present invention also provides a protein comprising amino acids in the sequence set forth by SEQ ID NO: 38 or SEQ ID NO: 39, except that: the amino acid at position 406 is an amino acid other than F; and/or the amino acid at position 474 is an amino acid other than N.

The present invention provides members that produce on a large scale a coenzyme-linked glucose dehydrogenase which has excellent substrate-recognizing ability toward glucose while providing low action on maltose. The present invention relates to a polynucleotide encoding a soluble coenzyme-linked glucose dehydrogenase that catalyzes the oxidation of glucose in the presence of an electron acceptor and has an activity toward maltose of 5% or lower; a polypeptide encoded by the nucleotide sequence of the polynucleotide; a recombinant vector carrying the polynucleotide; a transformed cell produced using the recombinant vector; a method for producing a polypeptide comprising culturing the transformed cell and collecting from the cultivated products a polypeptide that links to FAD to exert the glucose dehydration activity; a method for determination of glucose using the polypeptide; a reagent composition for determination of glucose; and a biosensor.

In some embodiments, the present invention a mutated FAD-GDH protein, wherein the mutated FAD-GDH protein is mutated from a wild-type first species to contain at least one point mutation, wherein the mutated FAD-GDH protein comprises: P(X).sub.n=8X.sup.4(X).sub.n=16V(X).sub.n=6RN(X).sub.n=3YDXRPXCXGX.sup.3NNCMP(X).sub.n=1CP(X).sub.n=2A(X).sub.n=1Y(X).sub.n=1G(X).sub.n=6A(X).sub.n=2AG(X).sub.n=6AVV(X).sub.n=3E(X).sub.n=8-9A(X).sub.n=2Y(X).sub.n=1D(X).sub.n=5HRV(X).sub.n=5V(X).sub.n=2A(X).sub.n=3E(X).sub.n=2K(X).sub.n=4S(X).sub.n=5P(X).sub.n=1G(X).sub.n=2N(X).sub.n=4GRN(X).sub.n=1MDH(X).sub.n=4V(X).sub.n=1F(X).sub.n=6-7W(X).sub.n=1GRGP(X).sub.n=9RDGXX.sup.5R(X).sub.n=19T(X).sub.n=14L(X).sub.n=14X.sup.2(X).sub.n=1X.sup.1(X).sub.n=1E(X).sub.n=4P(X).sub.n=1NR(X).sub.n=3S(X).sub.n=4D(X).sub.n=2G(X).sub.n=7Y(X).sub.n=4Y(X).sub.n=32-35, wherein each X represents a wild-type amino acid residue of the first species and n indicates the number of the wild-type amino acid residues of the first species represented by a respective parenthetical at that position, wherein: a) X.sup.1 is selected from the group consisting of X, S, C, T, M, V, Y, N, P, L, G, Q, A, I, D, W, H, and E, wherein if X.sup.1 is L, H or V, then X.sup.2 is D; b) X.sup.3 is selected from the group consisting of G, H, D, Y, S, and X; c) X.sup.4 is selected from the group consisting of S and X; and d) X.sup.5 is selected from the group consisting of L and X.

A recombinant protein, including: (a) alpha subunit of an FAD-GDH; and (b) a minimal cytochrome c peptide is provided. Additionally, an electrode coupled to a recombinant protein, the recombinant protein made of: (a) a cofactor of a redox enzyme; (b) a redox enzyme; (c) a linker moiety configured to link any one of: the cofactor or the enzyme to an electron transfer (ET) domain; and (d) an ET domain, is also provided. Methods for: (a) transferring an electron to an electrode, by coupling the recombinant protein an electrode; and (b) quantifying the amount of an analyte e.g., glucose are also provided.