Kao Corporation

Shinshu University

Katsuya Ozaki

Susumu Ito

Keiji Endo

Kazuaki Igarashi

Shuji Kawai

Hiroshi Hagihara

Yasuhiro Hayashi

Yuji Hatada

Katsutoshi Ara

Kikuhiko Okamoto

Shitsuw Shikata

Hiroyuki Araki

Kenzo Koike

Kazushi Oshino

Masatoshi Tohata

Kazuhisa Sawada

Naotake Ogasawara

Kaori Kitayama

Junichi Sekiguchi

Akira Takei

Shigeo Inoue

Tadahiro Ozawa

Tadashi Yoshimatsu

Hajime Mori

Hiromi Okoshi

Kaori Ikawa

Takeko Kodama

Tomokazu Sato

Yuichi Ota

Kazuo Kobayashi

The present invention provides a modified promoter DNA capable of enhancing transcription of genes encoding proteins or polypeptides, and a method for producing proteins or polypeptides efficiently by use of the modified promoter DNA. A promoter DNA recognized by SigA and SigE, which is produced by modifying a nucleotide sequence including a promoter recognized by SigA and bases in the vicinity thereof; an expression vector harboring the promoter DNA; a recombinant microorganism containing the expression vector; and a method for producing proteins or polypeptides characterized by culturing the recombinant microorganism.

Modified promoter

The present invention provides a mutantbacterium capable of enhancing productivity of proteins or polypeptides, a recombinant microorganism produced by introducing genes encoding heterologous proteins or polypeptides into the mutantbacterium, and a method for producing proteins or polypeptides by use of the recombinant microorganism. A mutantbacterium which has, on the genome or plasmid thereof, DNA having a promoter sequence which is recognized and transcribed specifically during the sporulation stage and which is ligated to an upstream end of a sigA gene or a gene equivalent thereto, a recombinant microorganism produced by introducing genes encoding heterologous proteins or polypeptides into the mutantbacterium, and a method for producing proteins or polypeptides by use of the recombinant microorganism.

[object Object]

A host microorganism capable of increasing productivity of a protein or polypeptide, a recombinant microorganism obtained by introducing a gene encoding a protein or polypeptide into the host microorganism, and a method for producing a protein or polypeptide using the recombinant microorganism are provided. Also provided is a recombinant microorganism obtained by introducing into a host microorganism a gene encoding a heterologous protein or polypeptide, wherein in said host microorganism theaprX gene or a gene corresponding to the aprX gene has been deleted or knocked out, and a method for producing a protein or polypeptide using the recombinant microorganism.

Recombinant microorganism comprising inactivation of the AprX gene

A microorganism wherein one or more genes selected from the group of genes participating in sporulation in the middle to late stages of sporulation have been deleted or inactivated; and a process for producing a target product (a protein) by use the microorganism. No spore is formed when this microorganism is employed, thereby enabling production of a target product (a protein) while decreasing energy loss, production of a by-product and specific production speed to decrease unnecessary consumption of a medium. Moreover, the production period can be prolonged, whereby the target product (the protein) can be produced efficiently.

Host microorganisms

The present invention provides a recombinant microorganism obtained by transferring, into a host microorganism which is capable of producing protein or polypeptide with increased productivity and which was identified by the present inventors, a gene encoding a protein or polypeptide, and a method for producing a protein or polypeptide by use of the recombinant microorganism.

Recombinant microorganism

The invention relates to mutant &#x3b1;-amylases that may be produced at high yield from recombinant microorganisms.

Highly productive &#x3b1;-amylases

The invention relates to a mutant &#x3b1;-amylase obtained by making replacement or deletion of at least one of amino acid residues such as the 167th Gln, 169th Tyr and 178th Ala in the amino acid sequence set forth in SEQ ID NO:1 in an &#x3b1;-amylase having said amino acid sequence, or an &#x3b1;-amylase having a homology of at least 70% to said amino acid sequence, a gene encoding the mutant &#x3b1;-amylase, a vector, transformed cells, a process for producing a mutant &#x3b1;-amylase, comprising culturing the transformed cells, and a detergent composition comprising the mutant &#x3b1;-amylase. The mutant &#x3b1;-amylase of the invention has excellent properties of high resistance to chelating agents, high specific activity in an alkaline region and excellent stability to heat, and is hence useful for detergents for automatic dish washer, laundry detergents and the like.

Mutant &#x3b1;-amylases

The present invention is directed to a liquefying alkaline alpha-amylase, and a DNA encoding for the same and functional fragments thereof.

Gene encoding alkaline liquefying &#x3b1;-amylase

Liquefying alkaline amylases, each having residual activity not less than 70% when treated at pH 10 and 45&#xb0; C. for 30 minutes in the presence of 1 to 100 mM of EDTA or EGTA, are described. Also described are detergents comprising these amylases. In comparison with conventional amylases for detergents, the liquefying alkaline amylases of this invention have a high chelating-agent resisting performance.

Amylases

Production process of glucose-1-phosphate

The invention relates to mutant &agr;-amylases that may be produced at high yield from recombinant microorganisms.

Highly productive alpha-amylases

The present invention provides: a nucleotide sequence encoding alkaline pullulanase and exhibiting both alkaline &agr;-amylase and alkaline pullulanase activity; a nucleotide sequence encoding alkaline &agr;-amylase possessing an amino acid sequence described in SEQ ID NO:3; a nucleotide sequence encoding alkaline pullulanase possessing an amino acid sequence described in SEQ ID NO:4; recombinant DNAs containing these nucleotide sequences, and transformed microorganisms harboring the recombinant DNAs. The technique of the present invention enables mass production of alkaline pullulanase exhibiting both alkaline &agr;-amylase and alkaline pullulanase activity.

Gene for enzyme having both alkaline pullulanase and alkaline alpha-amylase activities

Gene encoding alkaline liquifying alpha-amylase

The invention relates to a mutant &agr;-amylase having an amino acid sequence obtained by making deletion or replacement by another arbitrary amino acid residue of at least a methionine residue at the 202-position or a position homologous thereto among amino acid residues set forth in SEQ ID NO:1, which constitute a liquefying alkaline &agr;-amylase, a gene thereof, and a detergent composition comprising the mutant &agr;-amylase. The mutant &agr;-amylase has the optimum pH in an alkaline range, an excellent &agr;-amylase activity, and high and lasting resistance to oxidizing agents, and is hence particularly useful as a component of detergent compositions containing a bleaching agent and an oxidizing agent.

Mutant &agr;-amylases

Described are liquefying alkaline amylases each having residual activity not less than 70% when treated at pH 10 and 45&deg; C. for 30 minutes in the presence of 1 to 100 mM of EDTA or EGTA; and a detergent comprising the same. Compared with the conventional amylases for a detergent, they have high chelating-agent resisting performance.

Growing community of inventors

Tochigi, Japan

Katsuya Ozaki