Abstract: A method to efficiently extract and purify a target foreign protein expressed with the use of an E. coli strain from the periplasm is disclosed. The method for producing a protein comprising the steps (a) to (c) below: (a) a step of introducing a gene encoding a target foreign protein into E. coli having modification in a gene associated with maintenance of the outer membrane structure; (b) a step of culturing the E. coli of (a); and (c) a step of recovering a target foreign protein from the periplasmic fraction of the microbial cell after culture.

Abstract: A vector is provided that includes a nucleotide sequence selected from any one of (a) to (d). The selection of (a) to (d) includes: (a) the nucleotide sequence set forth in SEQ ID NO: 12, 15, 18, or 21, (b) the nucleotide sequence complementary to the nucleotide sequence set forth in SEQ ID NO: 12, 15, 18, or 21, where (a) and (b) hybridize under stringent conditions, (c) the nucleotide sequence having 90% or more sequence identity with the nucleotide sequence set forth in SEQ ID NO: 12, 15, 18, or 21, and (d) the nucleotide sequence set forth in SEQ ID NO: 12, 15, 18, or 21 in which a total of 1 to 50 nucleotides are substituted, deleted, or inserted.

Abstract: A polypeptide includes an amino acid sequence selected from the group consisting of: an amino acid sequence having at least 85% sequence identity to the amino acid sequence of SEQ ID NO: 1; and an amino acid sequence derived from the amino acid sequence of SEQ ID NO: 1 by substitution, deletion, and/or addition of one or more amino acid residues. A yeast host expressing the polypeptide in a secretory production system does not add an N-linked sugar chain to the polypeptide, and the polypeptide has endonuclease activity.

Abstract: It is an object of the present invention to provide a method for producing a low-molecular-weight antibody such as a Fab-type antibody, using yeast as a host, wherein the method is able to produce the low-molecular-weight antibody with high productivity. According to the present invention, there is provided a gene comprising a nucleotide sequence encoding an amino acid or an amino acid sequence capable of increasing the secretion amount of a Fab-type antibody at the 3?-terminus of a nucleotide sequence encoding the amino acid sequence of the Fd chain or L chain of an antibody.

Abstract: A polypeptide includes an amino acid sequence selected from the group consisting of: an amino acid sequence having at least 85% sequence identity to the amino acid sequence of SEQ ID NO: 1; and an amino acid sequence derived from the amino acid sequence of SEQ ID NO: 1 by substitution, deletion, and/or addition of one or more amino acid residues. A yeast host expressing the polypeptide in a secretory production system does not add an N-linked sugar chain to the polypeptide, and the polypeptide has endonuclease activity.

Abstract: The present invention has its object to provide a novel polypeptide having amidase activity to selectively hydrolyze S-enantiomer in racemic nipecotamide, a DNA encoding the polypeptide, a vector containing the DNA, a transformant transformed with the vector, and a method for producing an optically active carboxylic acid amide and an optically active carboxylic acid in which a racemic carboxylic acid amide is hydrolyzed with the polypeptide or the transformant.

Abstract: A vector includes a nucleotide sequence according to any one of the following (a) to (d): (a) a nucleotide sequence set forth in SEQ ID NO: 12, 15, 18, or 21; (b) a nucleotide sequence of a nucleic acid that hybridizes with a nucleic acid consisting of a nucleotide sequence complementary to the nucleotide sequence set forth in SEQ ID NO: 12, 15, 18, or 21, under stringent conditions; (c) a nucleotide sequence having 85% or more sequence identity with the nucleotide sequence set forth in SEQ ID NO: 12, 15, 18, or 21; (d) a nucleotide sequence set forth in SEQ ID NO: 12, 15, 18, or 21 in which one or more nucleotides are substituted, deleted, inserted, and/or added.

Abstract: The present invention relates to a method for producing an L-amino acid by reacting an enantiomeric mixture of an N-succinyl amino acid with L-succinylase in the presence of N-acylamino acid racemase to specifically hydrolyze the L-form. In particular, the present invention relates to a method for producing an L-amino acid in high yield by using an N-succinyl amino acid whose dissolved concentration is particularly low as a raw material to perform a reaction while precipitating the produced L-amino acid out of the reaction system. The present invention enables efficient production of an L-amino acid having high optical purity, particularly an L-amino acid useful as a raw material for products such as pharmaceutical products and agricultural chemicals.

Abstract: Water-forming NADH oxidase derived from Streptococcus mutans should be further improved in terms of stability for practical use in industrial production. An object of the present invention is to provide an enzyme that is obtained through modification of a water-forming NADH oxidase, which is useful as an NAD+ regeneration system for stereoselective oxidation catalyzed by an oxidoreductase, by protein engineering techniques so that the enzyme can withstand long-term use without exhibiting a reduction of its activity for the regeneration of NAD+, that is, an enzyme having improved stability, and to provide a method for efficiently producing a useful substance such as an optically active alcohol or amino acid. The present invention relates to an enzyme modification method that can improve the stability of water-forming NADH oxidase derived from Streptococcus mutans by appropriately introducing mutation.

Abstract: Water-forming NADH oxidase derived from Streptococcus mutans should be further improved in terms of stability for practical use in industrial production. An object of the present invention is to provide an enzyme that is obtained through modification of a water-forming NADH oxidase, which is useful as an NAD+ regeneration system for stereoselective oxidation catalyzed by an oxidoreductase, by protein engineering techniques so that the enzyme can withstand long-term use without exhibiting a reduction of its activity for the regeneration of NAD+, that is, an enzyme having improved stability, and to provide a method for efficiently producing a useful substance such as an optically active alcohol or amino acid. The present invention relates to an enzyme modification method that can improve the stability of water-forming NADH oxidase derived from Streptococcus mutans by appropriately introducing mutation.

Abstract: It is an object of the present invention to provide a method for producing a low-molecular-weight antibody such as a Fab-type antibody, using yeast as a host, wherein the method is able to produce the low-molecular-weight antibody with high productivity. According to the present invention, there is provided a gene comprising a nucleotide sequence encoding an amino acid or an amino acid sequence capable of increasing the secretion amount of a Fab-type antibody at the 3?-terminus of a nucleotide sequence encoding the amino acid sequence of the Fd chain or L chain of an antibody.

Abstract: Water-forming NADH oxidase derived from Streptococcus mutans should be further improved in terms of stability for practical use in industrial production. An object of the present invention is to provide an enzyme that is obtained through modification of a water-forming NADH oxidase, which is useful as an NAD+ regeneration system for stereoselective oxidation catalyzed by an oxidoreductase, by protein engineering techniques so that the enzyme can withstand long-term use without exhibiting a reduction of its activity for the regeneration of NAD+, that is, an enzyme having improved stability, and to provide a method for efficiently producing a useful substance such as an optically active alcohol or amino acid. The present invention relates to an enzyme modification method that can improve the stability of water-forming NADH oxidase derived from Streptococcus mutans by appropriately introducing mutation.

Abstract: An object of the present invention it to improve the secretion productivity of a heterologous protein in a methanol-assimilating yeast. The present invention provides a method for producing a heterologous protein, characterized by comprising using a methanol-assimilating yeast with disruption of the VPS gene as a host for secretory production of the heterologous protein.

Abstract: An object of the present invention it to improve the secretion productivity of a heterologous protein in a methanol-assimilating yeast. The present invention provides a method for producing a heterologous protein, characterized by comprising using a methanol-assimilating yeast with disruption of the VPS gene as a host for secretory production of the heterologous protein.

Abstract: The present invention has its object to provide a novel polypeptide having amidase activity to selectively hydrolyze S-enantiomer in racemic nipecotamide, a DNA encoding the polypeptide, a vector containing the DNA, a transformant transformed with the vector, and a method for producing an optically active carboxylic acid amide and an optically active carboxylic acid in which a racemic carboxylic acid amide is hydrolyzed with the polypeptide or the transformant.

Abstract: Water-forming NADH oxidase derived from Streptococcus mutans should be further improved in terms of stability for practical use in industrial production. An object of the present invention is to provide an enzyme that is obtained through modification of a water-forming NADH oxidase, which is useful as an NAD+ regeneration system for stereoselective oxidation catalyzed by an oxidoreductase, by protein engineering techniques so that the enzyme can withstand long-term use without exhibiting a reduction of its activity for the regeneration of NAD+, that is, an enzyme having improved stability, and to provide a method for efficiently producing a useful substance such as an optically active alcohol or amino acid. The present invention relates to an enzyme modification method that can improve the stability of water-forming NADH oxidase derived from Streptococcus mutans by appropriately introducing mutation.

Abstract: The present invention has its object to provide a method of producing an erythro- or threo-2-amino-3-hydroxypropionic acid ester, and so forth. The present invention relates to: a method of asymmetrically reducing an N-2-amino-3-oxopropionic acid ester by allowing cells of a microorganism to act thereon; a polypeptide having an activity of asymmetrically reducing a carbonyl compound to give an optically active alcohol, which is isolated from a microorganism belonging to genus Brevundimonas; a DNA coding for the polypeptide; and a transformant producing the polypeptide. The invention also relates to a method of producing an optically active alcohol by reducing a carbonyl compound with the help of the polypeptide or the transformant.

Abstract: An object of the present invention is to provide a novel alcohol dehydrogenase, a gene for the alcohol dehydrogenase, a vector including the gene, a transformant transformed with the vector, and a method for producing an optically active alcohol by utilizing them. A feature of the present invention directs to a novel polypeptide isolated from Candida maltosa, a DNA coding for the polypeptide, and a transformant producing the polypeptide. Another feature of the present invention directs to a method for producing an optically-active alcohol by reducing a carbonyl compound with the polypeptide or the transformant.

Abstract: The present invention relates to a method for producing an L-amino acid by reacting an enantiomeric mixture of an N-succinyl amino acid with L-succinylase in the presence of N-acylamino acid racemase to specifically hydrolyze the L-form. In particular, the present invention relates to a method for producing an L-amino acid in high yield by using an N-succinyl amino acid whose dissolved concentration is particularly low as a raw material to perform a reaction while precipitating the produced L-amino acid out of the reaction system. The present invention enables efficient production of an L-amino acid having high optical purity, particularly an L-amino acid useful as a raw material for products such as pharmaceutical products and agricultural chemicals.

Abstract: An object of the present invention is to provide a novel alcohol dehydrogenase, a gene for the alcohol dehydrogenase, a vector including the gene, a transformant transformed with the vector, and a method for producing an optically active alcohol by utilizing them. A feature of the present invention directs to a novel polypeptide isolated from Candida maltosa, a DNA coding for the polypeptide, and a transformant producing the polypeptide. Another feature of the present invention directs to a method for producing an optically-active alcohol by reducing a carbonyl compound with the polypeptide or the transformant.