Phytase and gene encoding said phytase
AssigneesKyowa Nakko Kogyo Co., Ltd., Shin Nihon Chemical Co., Ltd.
InventorsHidemasa Kondo, Hideharu Anazawa, Syunichi Kaneko, Tadashi Nagashima, Tatsuya Tange
Life of this patent
Estimating the term…
Abstract
This invention relates to an inexpensive phytase with a low Km value for phytic acid, DNA coding for the phytase, recombinant DNA having the DNA introduced thereinto, a transformant carrying the recombinant DNA, a process for preparing a phytase by use of the transformant, and an animal feed comprising the phytase.
Claims23
An isolated or purified gene coding for a phytase, from a microorganism belonging to the genus Aspergillus, having the following properties:
- (1) a Michaelis constant of 10 to 30 μM when phytic acid used as the substrate;
- (2) molecular weight as determined by SDS-PAGE:
- about 60 kDa after treatment with endoglycosidase H;
- (3) optimum pH:
- 5.0 to 6.5;
- (4) optimum temperature:
- 45° C. to 65° C;
- (5) acting on the substrates:
- phytic acid, p-nitrophenylphosphate, D-glucose 6-phosphate, fructose 6-phosphate, D-myo-inositol 1,4,5-tris-phosphate, glycerol phosphate, adenosine triphosphate;
- and
- (6) isoelectric focusing:
- pI 4.7 to 5.4.
Recombinant DNA obtained by introducing into a vector a DNA fragment containing a gene coding for a phytase, from a microorganism belonging to the genus Aspergillus, having the following properties:
- (1) a Michaelis constant of 10 to 30 μM when phytic acid used as the substrate;
- (2) molecular weight as determined by SDS-PAGE:
- about 60 kDa after treatment with endoglycosidase H;
- (3) optimum pH:
- 5.0 to 6.5;
- (4) optimum temperature:
- 45° C. to 65° C;
- (5) acting on the substrates:
- phytic acid, p-nitrophenylphosphate, D-glucose 6-phosphate, fructose 6-phosphate, D-myo-inositol 1,4,5-tris-phosphate, glycerol phosphate, adenosine triphosphate;
- and
- (6) isoelectric focusing:
- pI 4.7 to 5.4.
A transformant carrying recombinant DNA obtained by introducing into a vector a DNA fragment containing a gene coding for a phytase, from a microorganism belonging to the genus Aspergillus, having the following properties:
- (1) a Michaelis constant of 10 to 30 μM when phytic acid used as the substrate;
- (2) molecular weight as determined by SDS-PAGE:
- about 60 kDa after treatment with endoglycosidase H;
- (3) optimum pH:
- 5.0 to 6.5;
- (4) optimum temperature:
- 45° C. to 65° C;
- (5) acting on the substrates:
- phytic acid, p-nitrophenylphosphate, D-glucose 6-phosphate, fructose 6-phosphate, D-myo-inositol 1,4,5-tris-phosphate, glycerol phosphate, adenosine triphosphate;
- and
- (6) isoelectric focusing:
- pI 4.7 to 5.4.
A process for preparing a phytase from a microorganism belonging to the genus Aspergillus, wherein the microorganism is a transformant carrying recombinant DNA obtained by introducing into a vector a DNA fragment containing a gene coding for a phytase, from said microorganism, having the following properties:
- (1) a Michaelis constant of 10 to 30 μM when phytic acid used as the substrate;
- (2) molecular weight as determined by SDS-PAGE:
- about 60 kDa after treatment with endoglycosidase H;
- (3) optimum pH:
- 5.0 to 6.5;
- (4) optimum temperature:
- 45° C. to 65° C;
- (5) acting on the substrates:
- phytic acid, p-nitrophenylphosphate, D-glucose 6-phosphate, fructose 6-phosphate, D-myo-inositol 1,4,5-tris-phosphate, glycerol phosphate, adenosine triphosphate;
- and
- (6) isoelectric focusing:
- pI 4.7 to 5.4,
- wherein said process comprises culturing in a medium said microorganism, producing and accumulating phytase in said medium, and then recovering phytase from said medium.
Dependencies are inferred from the claim text.