Kao Corporation

Katsuhisa Saeki

Mitsuyoshi Okuda

Katsutoshi Ara

Kazuaki Igarashi

Tsuyoshi Sato

Masafumi Nomura

Yasushi Kageyama

Susumu Ito

Nobuyuki Sumitomo

Mikio Takaiwa

Tohru Kobayashi

Shitsuw Shikata

Hiromi Kubota

Jun Hitomi

Katsuhiko Deguchi

Shuji Kawai

Masaki Tosaka

Masatoshi Tohata

Hiroyuki Araki

Yuji Hatada

Yoshifumi Izawa

Taeko Sone

Akinori Ogawa

Kazuhiro Saito

Eiichi Hoshino

Keiji Endo

Hiroshi Hagihara

Atsushi Tanaka

Yasunao Wada

Yumi Nishimura

Takaaki Uemura

Providing an alkaline protease exhibiting an improved solubility in a liquid detergent. A mutant alkaline protease consisting of the amino acid sequence represented by SEQ ID No: 2 or an amino acid sequence having 80% or more sequence identity therewith, in which at least one amino acid residue selected from the group consisting of the amino acid residues at predetermined positions of the amino acid sequence represented by SEQ ID No: 2 or corresponding positions thereto are substituted.

Improving the solubility of an alkaline protease in a liquid detergent by amino acid substitution

The invention is directed to an alkaline protease that has the amino acid sequence of SEQ ID NO: 2 or of an amino acid sequence having an identity of 90% or more thereto, except that one or more amino acid residues at positions selected from (a) position 6, (b) position 15, (c) position 16, (d) position 65, (e) position 66, (f) position 82, (g) position 83, (h) position 204, (i) position 319, and (j) position 337 of SEQ ID NO: 2, or at positions corresponding thereto, are substituted with certain other amino acid residues. The invention is also directed to detergent compositions and culture medium that contain the protease.

Alkaline protease variants

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 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.

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 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.

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.

DNA fragment containing gene for alkaline pullulanase

Liquefying alkaline .alpha.-amylase, process for producing the same, and

A novel detergent composition containing an alkaline pullulanase is disclosed. The alkaline pullulanase has its optimum pH in an alkaline range and not deactivated by surfactants. Further it has strong resistance to almost all detergent components such as chelating agents, protease, etc. The the detergent composition of this invention has excellent detergency especially against starch soils.

Detergent composition containing alkaline pullylanase enzyme

A detergent composition containing an alkaline pullulanase, a surfactant, alkaline agents and/or inorganic electrolytes, divalent metal ion scavengers and bleaching agents is disclosed. The alkaline pullulanase has an optimum pH range of 8.5-10.0 on pullulan, an optimum temperature of about 50.degree. C. and is not deactivated by surfactants. Further, the pullulanase has a strong resistance to almost all detergent components such as chelating agents, proteases, etc. The pullulanase is isolated from Bacillus sp. KSM-AP 1378 deposited as FERM BP-3048. The composition specifically contains 0.1-10 wt. % alkaline pullulanase B, 0.5-60 wt. % surfactant, 0-90 wt. % alkaline agents and/or inorganic electrolytes, 0-50 wt. % divalent metal ion scavengers, and 0-85 wt. % bleaching agents.

Detergent composition containing an alkaline pullulanase from bacillus

An isolated alkaline pullulanase Y having .alpha.-amylase activity; a microorganism producing the alkaline pullulanase Y; and a process for producing the alkaline pullulanase Y are disclosed. The alkaline pullulanase Y having .alpha.-amylase activity has its optimum pH at higher alkaline range than conventional alkaline pullulanases and exhibits excellent stability in a wide pH range. Further, the alkaline pullulanase Y has strong resistance to almost all detergent components such as surfactants, chelating agents, proteases for detergents, and the like. Thus the alkaline pullulanase Y can advantageously be used as a detergent component.

Alkaline pullulanase Y having .alpha.-amylase activity

A novel alkaline pullulanase; a microorganism producing the alkaline pullulanase; and a process for producing the alkaline pullulanase are disclosed. The alkaline pullulanase of the present invention has a higher optimum pH range (pH 9.5-11) than conventional alkaline pullulanases and shows excellent stability in a wider pH range. It has also an optimum temperature of higher than 50.degree. C. and is thermally stable up to 40.degree. C. The alkaline pullulanase has also strong reistance to almost all detergent components such as surfactants, chelating agents, proteases for detergents, and the like. The alkaline pullulanase can advantageously be used as a detergent component. Also, it can be utilized for producing many kinds of oligosaccharides and also together with .alpha.-amylase for producing various monosaccharides.

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Haga-gun, Japan

Katsuhisa Saeki