Kaneka Corporation

Shinichi Yoshida

Dai Murata

Shunichi Taira

Naoaki Taoka

Keita Iguchi

Masayuki Takano

Tozo Nishiyama

Akira Iwasaki

Daisuke Moriyama

Yoshiyuki Nakano

Motohisa Washida

Shigeru Kawano

Shunsuke Sato

Shingo Kobayashi

Tomoyuki Nakaishi

Masutoshi Nojiri

Masahiro Hayashi

Fuminori Konoike

Kazunobu Minakuchi

Masakatsu Nishihachijyo

Junya Akagi

Yasuhiko Tabata

Yoshiyuki Tago

Kohji Ohno

Provided is a mutant PHA synthase which produces of a PHA copolymer with a high or low 3HH ratio while maintaining PHA productivity. The mutant PHA synthase is a mutant polyhydroxyalkanoate synthase having an amino acid sequence having 85% or more sequence identity with the amino acid sequence of SEQ ID NO: 1 and having at least one of the following mutations (a) to (c): (a) a substitution of serine at 389th position from N-terminus of the amino acid sequence of SEQ ID NO: 1 with an amino acid other than serine; (b) a substitution of leucine at 436th position from the N-terminus of the amino acid sequence of SEQ ID NO: 1 with an amino acid other than leucine; and (c) a deletion of 11 to 19 amino acid residues from the C-terminus of the amino acid sequence of SEQ ID NO: 1.

Mutant polyhydroxyalkanoate synthase, gene thereof and transformant, and method for producing polyhydroxyalkanoate

An affinity separation matrix includes a water-insoluble base material; and a ligand that is immobilized on the water-insoluble base material, wherein the ligand is an antibody &#x3ba; chain variable region-binding peptide comprising B5 domain of Protein L derived from312 strain or a part thereof.

Affinity separation matrix for purifying protein containing immunoglobulin K chain variable region

A method for purifying an antibody or an antibody fragment containing &#x3ba;-chain variable region includes adsorbing at least one of the antibody or the antibody fragment onto an affinity separation matrix by contacting a liquid sample with the affinity separation matrix, washing the affinity separation matrix to remove impurities, and separating the at least one of the antibody or the antibody fragment from the affinity separation matrix by using an acetate buffer. The liquid sample includes the at least one of the antibody or the antibody fragment. The affinity separation matrix includes a water-insoluble carrier and a ligand selected from the group consisting of Protein L, a variant of Protein L, a domain of Protein L, and a variant of the domain. The ligand is immobilized on the water-insoluble carrier.

Method for purifying antibody or antibody fragment containing &#x3ba;-chain variable region

A first immunoglobulin &#x3ba; chain variable region-binding peptide includes an amino acid sequence of SEQ ID NO: 21 with substitution of one or more amino acid residues at the 15position, the 16position, the 17position or the 18position, wherein an acid dissociation pH thereof is shifted to a neutral side. A second immunoglobulin &#x3ba; chain variable region-binding peptide further includes deletion, substitution and/or addition of 1-20 amino acid residues at positions other than the 15position, the 16position, the 17position and the 18position. A third immunoglobulin &#x3ba; chain variable region-binding peptide includes an amino acid sequence with a sequence identity of 80% or more to the amino acid sequence of the first immunoglobulin &#x3ba; chain variable region-binding peptide.

Mutant immunoglobulin K chain variable region-binding peptide

A first Fab region-binding peptide includes an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 5 with substitution of one or more amino acid residues at the 17position and the 36position, wherein an acid dissociation pH thereof is shifted to a neutral side. A second Fab region-binding peptide further includes deletion, substitution and/or addition of one or more amino acid residues at positions other than the 17position and the 36position. A third Fab region-binding peptide includes an amino acid sequence with a sequence identity of 80% or more to the amino acid sequence of the first Fab region-binding peptide.

Modified Fab region-binding peptide

An object of the present invention is to provide a Fab region-binding peptide having an excellent ability for binding to a Fab region of IgG, an affinity separation matrix having the peptide as a ligand, and a method for producing a Fab region-containing protein, the method using the affinity separation matrix. Further, another object of the present invention is to provide a DNA encoding for the peptide, a vector containing the DNA, and a transformant which has been transformed by the vector. The Fab region-binding peptide according to the present invention is characterized in having a mutation at a specific site in comparison with wild-type SpG-&#x3b2;1.

Fab region-binding peptide

The objective of the present invention is to provide a peptide of which both of binding forces to a Fc region and a Fab region are superior. In addition, the objective of the present invention is to provide a DNA which encodes the peptide, a vector which contains the DNA, and a transformant which is transformed by the vector. The problem can be solved by providing the peptide having the specific sequence.

Immunoglobulin G-binding peptide

An object of the present invention is to create a novel engineered Protein A ligand having better antibody dissociation properties in the acidic condition compared with known engineered Protein A ligands. The present invention provides a protein having an affinity for an immunoglobulin, including an amino acid sequence obtained by introducing, into an amino acid sequence derived from any of E, D, A, B and C domains of Protein A, at least one amino acid substitution at any one or more of amino acid residues corresponding to positions 31 to 37 of the A, B and C domains (positions 29 to 35 of the E domain, positions 34 to 40 of the D domain), which are conserved in all the domains, the protein having a lower affinity for an Fab region of an immunoglobulin than a protein having the amino acid sequence before introduction of the substitution.

Protein capable of binding specifically to immunoglobulin, and immunoglobulin-binding affinity ligand

An object of the present invention is to provide a technique to create novel engineered protein ligands that, when immobilized through a lysine residue (its side chain &#x3b5;-amino group) which allows for efficient immobilization to a carrier, show the optimum binding capacity and binding efficiency to a target molecule. The present invention provides an engineered protein having a sequence obtained by replacing all the lysine residues in Protein A, which is the most typical protein ligand, with other amino acids, and adding lysine at a terminal; and an affinity separation matrix in which such an engineered protein is immobilized on a water-insoluble carrier by reductive amination or the like. This affinity separation matrix is characterized by its high binding capacity to a target molecule even when the immobilized amount of the ligand is small.

Protein for affinity-separation matrix

An object of the present invention is to develop techniques to create novel engineered protein ligands that maximize the binding capacity and binding efficiency to a target molecule of affinity separation matrices on which the protein ligands are immobilized. The present invention provides protein ligands (variants) that can be immobilized on carriers in a manner shown in schematic FIG.()-(), as well as antibody affinity separation matrices obtained by immobilizing such a protein ligand on a water-insoluble carrier. The affinity separation matrices are characterized by their excellent binding capacity and binding efficiency to a target molecule.

Protein ligand for affinity isolation matrix

An object of the present invention is to provide a method for converting the coenzyme dependency of enzymes of the medium-chain dehydrogenase/reductase (MDR) family. A further object of the present invention is to provide enzyme variants of the MDR family whose coenzyme dependency is converted by the conversion method and a method for enzymatically producing optically active alcohols using the enzymes. The present inventors developed a novel enzyme conversion method for converting the coenzyme dependency of enzymes of the MDR family, rationally designed enzyme variants that are altered by the enzyme conversion method to be able to use NADPH as a coenzyme from a useful enzyme of the MDR family that uses NADH as a coenzyme, and actually provide variants having such an ability.

Enzyme function modification method and enzyme variant thereof

An object of the present invention is to create a novel engineered Protein A ligand having better antibody dissociation properties in the presence of an acid than conventional engineered Protein A ligands and a further object of the present invention is to create a novel engineered Protein A ligand having higher alkali resistance. The present invention is to provide a protein having an affinity for an immunoglobulin, including an amino acid sequence derived from any of E, D, A, B and C domains of Protein A, wherein at least one Gly residue in the amino acid sequence is replaced with an amino acid other than Ala, and the protein has a lower affinity for an Fab region of an immunoglobulin than a protein including an amino acid sequence in which the Gly residue is replaced with Ala. Also, the present invention is to provide the protein having an affinity for an immunoglobulin, which has improved chemical stability in an alkaline condition compared to the corresponding domain.

Protein having affinity for immunoglobulin, and immunoglobulin-binding affinity ligand

Water-forming NADH oxidase derived fromshould 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 fromby appropriately introducing mutation.

Protein having NADH and/or NADPH oxidase activity

Isolated DNA encoding protein having improved stability

An object of the present invention is to provide novel polypeptides that are capable of binding to an immunoglobulin and have high stability against alkali. The present invention relates to proteins having the amino acid sequence of SEQ ID No:1 or 2.

Growing community of inventors

Takasago, Japan

Shinichi Yoshida