Kao Corporation

Nobuyuki Sumitomo

Tsuyoshi Sato

Mitsuyoshi Okuda

Tohru Kobayashi

Yasushi Takimura

Katsuhisa Saeki

Masafumi Nomura

Yoshifumi Izawa

Hiroyuki Araki

Yuji Hatada

Yasushi Kageyama

Kazuhiro Saito

Akinori Ogawa

To provide a plasmid vector having high productivity which can be used for large scale production of an industrially valuable and useful protein, and a transformant using the plasmid vector. A plasmid vector having DNA which encodes a plasmid replication protein in which one or more amino acid residues that are selected from (a) position 48, (b) position 262, (c) position 149 and (d) position 198 in the amino acid sequence represented by SEQ ID NO: 2 are substituted with (a) Ala, Gly, Thr, Arg, Glu, Asn or Gln, (b) Gly, Ser, Thr, Cys or Val, (c) Asn, and (d) Glu, respectively.

Vector encoding a plasmid replication protein and use thereof

Provided in the present invention is an alkaline protease wherein an amino acid residue at (a) position 65, (b) position 101, (c) position 163, (d) position 170, (e) position 171, (f) position 273, (g) position 320, (h) position 359 or (i) position 387 of SEQ. ID NO:1 or at a position corresponding thereto has been selected from the following amino acid residues: position (a): proline, position (b): asparagine, position (c): histidine, aspartic acid, phenylalanine, lysine, asparagine, serine, isoleucine, leucine, glutamine, threonine and valine, position (d): valine and leucine, position (e): alanine, glutamic acid, glycine and threonine, position (f): isoleucine, glycine and threonine, position (g): phenylalanine, valine, threonine, leucine, isoleucine and glycine, position (h): serine, leucine, valine, isoleucine and glutamine, position (i): alanine, lysine, glutamine, glutamic acid, arginine and histidine. The present invention makes it possible to efficiently produce and provide alkaline proteases having activity even in the presence of a highly concentrated fatty acid, and exhibiting excellent detergency for the removal of a complex stain containing protein, sebum and the like, and therefore being useful as an enzyme to be incorporated in a detergent.

Alkaline protease

The present invention provides an alkaline protease exhibiting high detergency and productivity, which is obtained by further enhancing the detergency of an alkaline protease effective against complex dirt and enhancing the specific activity.

An alkaline protease having an amino acid with one or more amino acid residues selected from among those located at (a) position 15, (b) position 16, (c) position 166, (d) position 167, (e) position 187, (f) position 346, and (g) position 405 of the amino acid sequence of SEQ No; 1, or at positions corresponding to these positions, are the following amino acid residues, respectively: (a): histidine, (b): threonine or glutamine, (c): glycine, (d): valine, (e): serine, (f): arginine, and (g); and a gene encoding the alkaline protease.

The present invention relates to alkaline proteases having high specific activity and strong oxidant resistance. The present invention also relates to alkaline proteases having excellent detergency that are to be added to a detergent.

Alkaline proteases

An alkaline protease wherein an amino acid residue at (a) position 65, (b) position 101, (c) position 163, (d) position 17-0, (e) position 171, (f) position 273, (g) position 320, (h) position 359 or (i) position 387 of SEQ. ID NO: 2 or at a position corresponding thereto has been selected from the following amino acid residues: position (a): proline, position (b): asparagine, position (e): histidine, aspartic acid, phenylalanine, lysine, asparagine, seine, isoleucine, leucine, glutamine, threonine and valine, position (d): valine and leucine, position (e): alanine, glutamic acid, glycine and threonine, position (K): isoleucine, glycine and threonine, position (g): phenylalanine, valine, threonine, leucine, isoleucine and glycine, position (h): seine, leucine, valine, isoleucine and glutamine, position (i): alanine, lysine, glutamine, glutamic acid, arginine and histidine. A method to an alkaline protease having activity even in the presence of a highly concentrated fatty acid, and exhibiting excellent detergency for the removal of a complex stain containing protein, sebum and the like, and therefore useful as an ingredient in a detergent.

The present invention relates to genes expressing an alkaline protease having a prepro sequence, vectors containing the same, and transformants containing said vector. The present invention also relates to a method for producing an alkaline protease.

Nucleic acid sequences encoding modified prepro peptides of an alkaline protease

The present invention relates to an alkaline protease having a prepro sequence, wherein the prepro sequence is a mutant sequence of the amino acid sequence of SEQ ID NO: 1 or an amino acid sequence having 80% or higher homology to the amino acid sequence of SEQ ID NO: 1, in which amino acid residues at (a) position 52, (b) position 75, and (c) position 142 of SEQ ID NO: 1, or amino acid residues at positions corresponding to these positions are substituted by the following amino acid residues: According to the present invention, an alkaline protease increased in production can be produced. In particular, there can be efficiently produced an alkaline protease having an activity even in the presence of a fatty acid of high concentration and exhibiting excellent detergency against complex soils containing proteins and sebum.

The present invention relates to alkaline proteases having high specific activity and strong oxidant resistance. The present invention also relates to alkaline proteases having excellent detergency that is to be added to a detergent.

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Haga-gun, Japan

Nobuyuki Sumitomo