Broad-spectrum, high-temperatur-resistant, high-specific-activity phytase, and its coding gene and expression

A technology of phytase and acid enzyme, which is applied in the direction of plant genetic improvement, genetic engineering, botany equipment and methods, etc., can solve the problems of waste of phosphorus sources, increase of feed cost, pollution of the environment, etc.

Inactive Publication Date: 2003-03-26
FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES +1
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0004] Phytate phosphorus is the basic storage form of phosphorus and inositol in the seeds of crops such as cereals, beans and oilseeds, with a content of up to 1-5% (Lolas M.Et al. Food Sci.42:1094-1097, 1977). It accounts for 60-80% of total phosphorus in plants (Nelson T.S.Poultry Sci.47:862-871, 1967). However, phosphorus in the form of phytate phosphorus is difficult to be utilized due to the lack of enzymes that can decompose phytic acid in monogastric animals (Cromwell G.L.Biotechnology in the Feed Industry, Lyons T.P.ed.AlltechTechnical Publication, Nicholasville, KY, 133-145), its utilization rate is only in 0-40%
[0005] Phytate phosphorus cannot be effectively utilized by monogastric animals, which causes many problems in the feeding process. First, it causes waste of phosphorus sources
On the one hand, the phosphorus source in the feed cannot be effectively utilized; on the other hand, in order to meet the animal’s demand for phosphorus, additional inorganic phosphorus must be added to the feed, which increases the feed co

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  • Broad-spectrum, high-temperatur-resistant, high-specific-activity phytase, and its coding gene and expression
  • Broad-spectrum, high-temperatur-resistant, high-specific-activity phytase, and its coding gene and expression
  • Broad-spectrum, high-temperatur-resistant, high-specific-activity phytase, and its coding gene and expression

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Embodiment Construction

[0047] experiment one

[0048] 1. Strain and carrier Escherichia coli strain E.coli DH 5a , plasmid pUC18, etc. were purchased from Promega, and yeast strain Pichia pastoris GS115 (His - Mut + ), plasmid pPIC9 was kindly donated by Dr. D.Luo, University of Alberta, Canada.

[0049] 2. Enzymes and kits Restriction enzymes, ligases, Taq enzymes, and Mung bean enzymes are products of Boehringer. T 7 DNA sequence kit was purchased from Pharmacia. In vitromutagenesis systems Kit, random primer marker Kit and PCR Kit were purchased from Promega.

[0050] 3. Biochemical reagents DNA synthesis reagents are products of Milipore Company. Primers were synthesized with ABI Cyclone DNA synthesizer. IPTG, X-Gal, SDS and sodium phytate are products of Sigma. TEMED, ammonium persulfate, acrylamide and methylene bisacrylamide are products of Promega.

[0051] 4. Substratum Aspergillus niger growth medium is PDA (20% potato, 2% sucrose, 2% agar); Aspergillus nige...

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Abstract

The invention provides a kind of broad-spectrum embedded acid enzyme and its coding gene. The invention also provides the expressing carrier and the recomposing yeast cell which contains the carrier. In the invention, the expressing quantity of embedded acid enzyme in recomposing yeast reaches to 520000U/ml ferment liquor.

Description

technical field [0001] The invention relates to a phytase and its coding gene. The invention also relates to an expression vector containing the modified phytase and a recombinant yeast cell containing the expression vector. Background technique [0002] Phosphorus is an essential mineral that all animals need for growth. Phosphorus accounts for more than a quarter of the total minerals in an animal's body, and 80% of it exists in the bones in the form of hydroxyapatite. Bones serve not only as the skeleton of the body, but also as a storehouse of calcium, phosphorus, and other minerals that can be mobilized to other tissues when needed. The other 20% of phosphorus exists in muscle, blood and other cartilage tissues. They are the constituents of many organic compounds and participate in almost every physiological and biochemical reaction required by life in the body. Because phosphorus has such important biological functions, it is very important to provide adequate phosp...

Claims

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Application Information

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IPC IPC(8): A23K10/16C12N1/19C12N9/16C12N15/55C12N15/63C12N15/81
Inventor 姚斌范云六王亚茹操时树袁铁铮史秀云
Owner FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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