AMANO ENZYME INC.

An object of the present invention is to find a new mutation effective for improving transglutaminase, and to provide a highly useful modified transglutaminase. Disclosed is a highly useful modified transglutaminase having an amino acid substitution that results in a reduction in temperature stability, an improvement in heat resistance, an improvement in oxidation resistance, an improvement in reactivity, or conversion into deamidase.

The present invention provides any one of the following modified esterases (i) to (iii): (i) a modified esterase comprising an amino acid sequence in which at least one amino acid at a site selected from the group consisting of (1) to (24) below in the amino acid sequence of SEQ ID NO: 1 or 3 is substituted with another amino acid, (1) V64 (2) S65 (3) A102 (4) A109 (5) L151 (6) I153 (7) Y173 (8) Y197 (9) L200 (10) I211 (11) F217 (12) S220 (13) D222 (14) D227 (15) L229 (16) L242 (17) A245 (18) V257 (19) L298 (20) T301 (21) V326 (22) S329 (23) A350 (24) T371; (ii) a modified esterase in which one or more amino acids are further substituted (except for the amino acid site substituted in (i)), added, inserted, or deleted (except for the amino acid site substituted in (i)) in the modified esterase of (i), and the reactivity to (1R,3R)-methyl 3-(2-chloro-3,3,3-trifluoroprop-1-en-1-yl)-2,2-dimethylcyclopropane-1-carboxylate is improved compared to the reactivity of an esterase consisting of the amino acid sequence of SEQ ID NO: 1 or 3; (iii) a modified esterase in which an amino acid(s) other than the amino acid(s) substituted in (i) is further substituted with another amino acid(s), wherein the modified esterase has 70% or more identity with the modified esterase of (i) and has improved reactivity to (1R,3R)-methyl 3-(2-chloro-3,3,3-trifluoroprop-1-en-1-yl)-2,2-dimethylcyclopropane-1-carboxylate compared to the reactivity of the esterase consisting of the amino acid sequence of SEQ ID NO: 1 or 3.

It is an object of the present invention to provide a modified oxygenase that specifically epoxidizes only the position-ω3 of highly unsaturated fatty acid. The present invention relates to: a polypeptide consisting of an amino acid sequence containing one or two or more amino acid substitutions selected from the group consisting of F87K/I/H, A330V, P25L and T438M, in the amino acid sequence as set forth in SEQ ID No: 2; and a polypeptide substantially identical to the aforementioned polypeptide.

The purpose of the present invention is to provide a modified transglutaminase that has improved reactivity in a heating temperature range. A transglutaminase, which includes a polypeptide having an amino acid sequence derived from an amino acid sequence represented by SEQ ID NO: 1 by: substitution of the combination of the three amino acid residues at the 249 to 251 positions by a combination of three predetermined amino acid residues; substitution of the combination of the three amino acid residues at the 243 to 245 positions by a combination of three predetermined amino acid residues; and/or substitution of the amino acid residue at the 2 position by a predetermined amino acid, has improved reactivity in a heating temperature range compared to the polypeptide having the amino acid sequence represented by SEQ ID NO: 1.

The object of the present invention is to provide a processing method that makes it possible to improve various characteristics related to a fermented food or beverage that uses a protein ingredient (and, in particular, characteristics of the fermented food or beverage itself such as stress, water retention, suppression of water release, and the like or characteristics of the ingredients of the fermented food or beverage related to fermentation speed during production of the fermented food or beverage). The present invention improves various characteristics of a fermented food or beverage that uses a protein ingredient using the action of a protein deamidase and a multicopper oxidase during production of the fermented food or beverage.

The present invention provides a liquid enzyme preparation of an enzyme having excellent activity stability. Stability of enzymatic activity is improved by accommodating a liquid enzyme preparation that includes an enzyme into a container the internal space of which has been replaced by an inert gas, or by adding, to a liquid enzyme preparation containing a protein deamidase, an additive selected from the group consisting of sugar alcohols, glycerin, ethanol, sulfites, lecithins, citrates, polyphosphates, tartrates, phytic acid, metaphosphoric acid, sodium chloride, tocotrienols, methionine, pectins, beet pigment, capsicum pigment, rosemary extract, tocopherols, tea extract, and butylated hydroxyanisole.

The present invention has a purpose of providing a β-galactosidase enzyme useful for the production of oligosaccharides. Disclosed is a β-galactosidase enzyme comprising the amino acid sequence of any one of SEQ ID NOs: 1 to 4 or an amino acid sequence that is 80% or more identical to said amino acid sequence.

Provided is a heat-resistant D-allulose-3-epimerase. A polypeptide shown in any one of the following (1) to (3): (1) a polypeptide consisting of an amino acid sequence having one or two or more amino acid substitutions selected from the group consisting of A56E, N57H, R58G and F111W in SEQ ID NO:1; (2) a polypeptide consisting of the amino acid sequence having substitution, addition, insertion or deletion of one or several amino acid residues in the amino acid sequence described in (1) above, having D-allulose-3-epimerase activity and retaining one or two or more amino acid substitutions selected from the group consisting of A56E, N57H, R58G, and F111W; and (3) a polypeptide having a 90% or more sequence identity to the amino acid sequence described in (1) above, having D-allulose-3-epimerase activity and retaining one or two or more amino acid substitutions selected from the group consisting of A56E, N57H, R58G, and F111W.

It is an object of the present invention to provide an enzyme agent for transesterification, including, as an active ingredient, a lipase having excellent heat resistance and high specificity for the 1,3-positions. According to the present invention, provided is an enzyme agent for transesterification, including, as an active ingredient, a lipase consisting of an amino acid sequence having a sequence identity of 90% or more to the amino acid sequence as set forth in SEQ ID NO: 2 or SEQ ID NO: 4.

The usefulness of β-galactosidases derived from Bacillus circulans is further enhanced. A modified β-galactosidase in which one or more amino acids selected from the group consisting of proline 182 (P182), tyrosine 187 (Y187), serine 188 (S188), tryptophan 405 (W405), alanine 406 (A406), glutamine 407 (Q407), tyrosine 449 (Y449), threonine 483 (T483), serine 512 (S512), serine 531 (S531), threonine 533 (T533), serine 534 (S534), asparagine 550 (N550), glutamine 551 (Q551), tryptophan 593 (W593), tyrosine 598 (Y598), proline 602 (P602), proline 604 (P604), tyrosine 609 (Y609), lysine 612 (K612), and tyrosine 615 (Y615), or an amino acid(s) corresponding thereto, has/have been substituted by other amino acid in a β-galactosidase consisting of the amino acid sequence of any of SEQ ID NOs. 1 to 4 or an amino acid sequence having 90% or more identity to the amino acid sequence set forth in any of SEQ ID NOs. 1 to 4.