Kao Corporation

Masafumi Nomura

Mitsuyoshi Okuda

Katsuhisa Saeki

Shitsuw Shikata

Tsuyoshi Sato

Tohru Kobayashi

Nobuyuki Sumitomo

Hiromi Kubota

Jun Hitomi

Yasushi Kageyama

Mikio Takaiwa

Nobuyuki Ogura

Yasushi Takimura

Hitoshi Tanimoto

Yoshifumi Izawa

Toshihiro Oki

Kazuhiro Saito

Tsutomu Tokumoto

Eiichi Hoshino

Atsushi Tanaka

Shitsuu Shikata

Yoko Ishii

The invention provides an activation method of &#x3b1;-amylase. &#x3b1;-Amylase or particles containing the same are contacted with an oxidizing agent or particles containing the same. After contacting, the activated &#x3b1;-amylase can be separated from the oxidizing agent. After contacting, &#x3b1;-amylase and the oxidizing agent can be incorporated in the form of a mixture into a detergent base to give a detergent composition.

Method of activating &#x3b1;-amylase with oxidizing agents

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

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.

An alkaline protease having the following properties; a gene encoding the same; a microorganism producing the same; and washing compositions containing the same; (i) acting over a broad pH value range of 4 to 13 and achieving, at pH 6 to 12, 80% or more the activity at the optimum pH value; (ii) when treated at 40&#xb0; C. for 30 minutes, being stable over a pH value range of 6 to 11; (iii) having an isoelectric point of about 8.9 to 9.1; and (iv) having casein digesting activity that is not inhibited by oleic acid. The alkaline protease of the present invention is highly stable to various surface active agents and fatty acids, and exhibits high stability to oxidizing agents, and is therefore useful as an enzyme to be used in detergents for automatic dishwashers and laundry detergents, both containing bleaching components.

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.

An alkaline protease having the following properties; a gene encoding the same; a microorganism producing the same; and washing compositions containing the same; (i) acting over a broad pH value range of 4 to 13 and achieving, at pH 6 to 12, 80% or more the activity at the optimum pH value; (ii) when treated at 40&deg; C. for 30 minutes, being stable over a pH value range of 6 to 11; (iii) having an isoelectric point of about 8.9 to 9.1; and (iv) having casein digesting activity that is not inhibited by oleic acid. The alkaline protease of the present invention is highly stable to various surface active agents and fatty acids, and exhibits high stability to oxidizing agents, and is therefore useful as an enzyme to be used in detergents for automatic dishwashers and laundry detergents, both containing bleaching components.

The present invention provides a detergent composition which is excellent in enzyme stability and exhibits excellent detergency particularly to protein-related dirt of socks and other items even under laundering conditions at a lower temperature. That is, the present invention provides a detergent composition comprising specific proportions of (a) an anionic surfactant, (b) a chlorine scavenger, (c) a protease whose &agr;-keratin-hydrolyzing activity at 10&deg; C. is not less than 0.09&times;10 &mgr;g/mPU&middot;min and (d) a protease whose &agr;-keratin-hydrolyzing activity at 10&deg; C. is less than 0.09&times;10 &mgr;g/mPU&middot;min.

Detergent composition

The present invention provides a detergent composition which is excellent in enzyme stability and exhibits excellent detergency particularly to protein-related dirt of socks and other items even under laundering conditions at a lower temperature. That is, the present invention provides a detergent composition comprising specific proportions of (a) an anionic surfactant, (b) a chlorine scavenger, (c) a protease whose &agr;-keratin-hydrolyzing activity at 10&deg; C. is not less than 0.09&times;10,&mgr;g/mPU&middot;min and (d) a protease whose &agr;-keratin-hydrolyzing activity at 10&deg; C. is less than 0.09&times;10,&mgr;g/mPU&middot;min.

A powdered detergent composition comprising a nonionic surfactant as the main surfactant, which comprises (a) a polyoxyethylene alkyl ether, wherein the alkyl group has 10 to 20 carbon atoms on the average and the average molar number of ethylene oxide added is 5 to 15, in an amount of 12 to 35% by weight based on the total weight of the composition, (b) a lipolytic enzyme in such an amount that the lipolytic activity per gram of component (a) would be 0.1 to 500 LU, (c) an anionic surfactant in an amount of 1.0 to 7.0% by weight based on the total weight of the composition and (d) an alkaline material in an amount of 5 to 35% by weight based on the total weight of the composition, and of which a 0.1% by weight aqueous solution has a pH exceeding 9.0, is excellent in detergency on oily soils such as subum soil.

Growing community of inventors

Wakayama, Japan

Masafumi Nomura