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Process of producing heat resistant xylanase with genetically recombinant Pichia yeast

A technology for xylanase and Pichia pastoris, which is applied in the field of gene recombination Pichia pastoris to produce high temperature resistant xylanase, can solve the problems of lack of heat resistance properties and the like, and achieve the effect of good heat resistance properties

Inactive Publication Date: 2007-11-07
YUNNAN NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

During this period, many domestic research institutes and companies began to carry out research work in this area, and achieved a large number of research results. Some products have entered the market, but the current products, whether domestic or foreign, do not have heat resistance.

Method used

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  • Process of producing heat resistant xylanase with genetically recombinant Pichia yeast
  • Process of producing heat resistant xylanase with genetically recombinant Pichia yeast
  • Process of producing heat resistant xylanase with genetically recombinant Pichia yeast

Examples

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Embodiment 1

[0054] 1. Construction of engineering bacteria

[0055] (1) Extraction of Aspergillus niger genomic DNA: For the extraction method, see Yao Bin et al. Screening of Aspergillus niger strains producing phytase and cloning of phytase gene, Journal of Agricultural Biotechnology, 1998, 6(1): 1-6

[0056] (2) Using PCR reaction to clone DNA gene fragments from Aspergillus niger genomic DNA: For the specific PCR method, see "CW Dieffenbach, PCR Technology Experiment Guide, Science Press" and , primers and reaction conditions are as follows:

[0057] Primer 1 (F): 5′-cggaattcgctcctgtgccggaacctg-3′

[0058] Primer 2 (R): 5′-cggaattcagaggagatcgtgacactggcgcg-3′

[0059] The reaction conditions were: denaturation at 94°C for 1 min, renaturation at 50°C for 1 min and 30 s, extension at 72°C for 2 min and 30 s, and after 30 cycles, incubation at 72°C for 20 min.

[0060] It is also possible to synthesize the whole gene by TaKaRa Dalian Company, the Aspergillus niger xylanase gene (as show...

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Abstract

The present invention is process of producing heat resistant xylanase with genetically recombinant Pichia yeast. Xylanase gene is separated from Aspergillus niger var niger strain N402 and expressed effectively in a eukaryon expression system Pichia yeast. The genetic engineering yeast constructed with Aspergillus niger var niger strain N402 xylanase gene, Pichia yeast expressing vector PGAP Z alpha A and host strain Pichia pastoris SMD1168 and GS115 has capacity of producing heat resistant xylanase. The heat resistant xylanase produced with the genetically recombinant Pichia yeast has excellent heat resistance optimal acting temperature of 37-50 deg.c and excellent catalytic effect in the stomach of animal, and is especially suitable for use in feed industry.

Description

technical field [0001] The invention relates to a method for producing high-temperature resistant xylanase by genetically recombined Pichia pastoris. Background technique [0002] Xylan is an important component of plant hemicellulose. It mainly exists in the cell wall of plants, accounting for about 35% of the dry weight of cells. It is the most abundant polysaccharide in nature except cellulose, and it is also a huge renewable resource. . However, xylan in wheat (such as wheat, rye, rice, bran, rice bran, etc.) feed, as a non-starch polysaccharide anti-nutritional factor, has a strong anti-nutritional effect on the digestion and absorption of animals. The main manifestations are: viscous combination with nutrients in the gastrointestinal tract of animals, reducing the absorption of nutrients; combining with digestive enzymes and bile acids, affecting digestion, increasing the amount of defecation, increasing sticky feces, and polluting the environment; leading to the prol...

Claims

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Application Information

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IPC IPC(8): C12N1/19C12N9/24C12N15/56C12R1/84
Inventor 黄遵锡许波唐湘华
Owner YUNNAN NORMAL UNIV
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