Kao Corporation

Kazuaki Igarashi

Katsuya Ozaki

Hiroshi Hagihara

Katsutoshi Ara

Susumu Ito

Katsuhisa Saeki

Yasuhiro Hayashi

Keiji Endo

Shuji Kawai

Norihiko Higaki

Hiroyuki Araki

Yuji Hatada

Shuichi Takizawa

Katsuhiko Deguchi

Masaki Tosaka

Jun Hitomi

Taeko Sone

Kaori Kitayama

Eiichi Hoshino

Tohru Kobayashi

Mikio Takaiwa

Atsushi Tanaka

Kaori Ikawa

Shuuichi Takizawa

Takaaki Uemura

This invention relates to a novel microorganism that efficiently produces a dodecahydro-3a,6,6,9a-tetramethylnaphtho[2,1-b]furan intermediate using sclareol as a substrate. As a result of concentrated studies, a plurality of novel microorganisms having properties of interest that are not classified as conventional microorganisms were isolated and identified. The novel microorganism of the present invention belongs toand has the ability of producing a dodecahydro-3a,6,6,9a-tetramethylnaphtho[2,1-b]furan intermediate using sclareol as a substrate. Such microorganism ofrepresents a new finding and it can be effective for producing dodecahydro-3a,6,6,9a-tetramethylnaphtho[2,1-b]furan and an intermediate thereof.

Microorganism and method for producing dodecahydro-3a,6,6,9a-tetramethylnaphtho[2,1-b]furan intermediate using the novel microorganism

This invention relates to a novel microorganism that efficiently produces a dodecahydro-3a,6,6,9a-tetramethylnaphtho[2,1-b]furan intermediate using sclareol as a substrate. As a result of concentrated studies, a plurality of novel microorganisms having properties of interest that are not classified as conventional microorganisms were isolated and identified. The novel microorganism of the present invention belongs to Ascomycetes and has the ability of producing a dodecahydro-3a,6,6,9a-tetramethylnaphtho[2,1-b]furan intermediate using sclareol as a substrate. Such microorganism of Ascomycetes represents a new finding and it can be effective for producing dodecahydro-3a,6,6,9a-tetramethylnaphtho[2,1-b]furan and an intermediate thereof.

It is intended to provide a process for conveniently producing phosphorylase at a high purity using a phosphorylase-producing microorganism. Namely, a process for producing phosphorylase characterized by culturing a phosphorylase-producing microorganism in a medium containing phosphoric acid or its salt at a concentration of 50 mM or above and collecting the phosphorylase thus produced in the medium.

Process for producing phosphorylase

The present invention provides an M-1-P-producing enzyme which employs, as starting materials, an oligosaccharide or polysaccharide having a glucose polymerization degree of 5 or more and containing an &#x3b1;-1,4-glycosidic bond such as maltooligosaccharide, dextrin, or starch, and which enables production of a large amount of M-1-P. The M-1-P-producing enzyme can produce M-1-P from a phosphoric acid or a salt thereof and an oligosaccharide or polysaccharide having a glucose polymerization degree of 5 or more and containing an &#x3b1;-1,4-glycosidic bond.

Maltose-1-phosphate-producing enzyme

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

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

DNA fragment containing gene for alkaline pullulanase

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.

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

Growing community of inventors

Tochigi, Japan

Kazuaki Igarashi