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Immobilization method and application of pichia pastoris

A technology of Pichia pastoris and immobilization method, which is applied in the field of Pichia pastoris immobilization, can solve problems affecting fermentation performance, etc., and achieve the effects of stable process, uniform distribution and high porosity

Active Publication Date: 2020-04-28
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using porous fiber materials as fixing materials not only has stable process, non-toxic, cheap and easy to obtain, good adsorption effect, high reuse rate, but also has a good protective effect on cell activity, but because the surface of Pichia pastoris is relatively hydrophilic, and With a negative charge, it is not easy to be fixed by the fiber medium during the fermentation process, which seriously affects the fermentation performance, so few people use the adsorption method as the fixation method for Pichia pastoris

Method used

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  • Immobilization method and application of pichia pastoris
  • Immobilization method and application of pichia pastoris
  • Immobilization method and application of pichia pastoris

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Experimental program
Comparison scheme
Effect test

Embodiment approach

[0053] (1) Wash the fiber material in deionized water for 1 to 3 hours, then soak the cleaned material in 10 to 50% (mass fraction) NaOH aqueous solution for 1 to 6 hours, and then wash the material in deionized water until 1 to 3 hours, until the NaOH is cleaned and the surface of the carrier is neutral;

[0054] (2) Place the porous fiber material treated in step (1) in 0.1 to 10% (volume fraction) vegetable glue aqueous solution, soak for 10 to 60 minutes, then place the porous fiber material in deionized water after fully rinsing Dry at 120°C and store for future use. Wherein, the vegetable gum is any one or more of squash gum, guar gum, flax gum, and cougar gum.

Embodiment 1

[0055] Embodiment 1: the preparation of porous fibrous material immobilization medium

[0056] (1) Wash the polyester fiber in deionized water for 1 h, then soak it in 25% (mass fraction) NaOH aqueous solution for 3 h, then wash the polyester fiber in deionized water for 3 h until the NaOH is cleaned, and the carrier surface neutral

[0057] (2) Soak the polyester fibers in 6% (volume fraction) water solution of squash gum for 30 minutes, then rinse the polyester fibers in deionized water, dry them at 120°C, and store them at 4°C for later use.

Embodiment 2

[0058] Embodiment 2: the preparation of porous fibrous material immobilization medium

[0059] (1) Wash the polyethylene carrier in deionized water for 1.5h, then place it in 25% (mass fraction) NaOH aqueous solution and soak it for 2h, then wash the polyethylene carrier in deionized water for 2h until the NaOH is cleaned, and the carrier surface neutral

[0060] (2) Soak the polyethylene in 3% (volume fraction) coumarin aqueous solution for 20 minutes, then fully rinse the polyethylene in deionized water, dry it at 120°C, and store it at 4°C for later use.

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Abstract

The invention discloses an immobilization method and application of pichia pastoris. Fibrous materials after being pretreated by plant glue are used as an immobilization carrier, the pichia pastoris is activated, then immobilization fermentation culture mediums are inoculated with the activated pichia pastoris, culture is performed, strains are adsorbed on the carrier in the culture process. In addition, the fibrous materials after being pretreated by the plant glue can be soaked in a methyl acryloxyethyl trimethyl ammonium chloride aqueous solution for secondary pretreatment, then washing anddrying are performed, so that the pichia pastoris is obtained. The immobilization method has the beneficial effects that through treatment with methyl acryloxyethyl trimethyl ammonium chloride, the surface of the carrier is provided with positive charges; the surfaces of materials are accessed with the positive charges of appropriate density, so that adsorption of proteins adhered on cell surfaces can be effectively strengthened, cell adhesion can be promoted, and besides, negative influence cannot be produced on the activity of cells; the enzyme activity of products is increased, the enzymeactivity of phytase produced through a fermenter is increased from 12326.85U / mL to 15221.85U / mL; and the whole fermentation cycle can be shortened.

Description

technical field [0001] The invention belongs to the technical field of industrial biology, and in particular relates to an immobilization method of Pichia pastoris and its application. Background technique [0002] The chemical name of phytic acid is inositol hexaphosphate (myo-inositolhexakisdihydrogenph-oshate), which is the main form of inositol and phosphoric acid in plant seeds. 40% to 70% of the total phosphorus. Phosphorus is one of the most functional inorganic nutrients in animals. In the process of animal feeding, sufficient phosphorus must be supplied in the feed to maintain the normal life and production activities of poultry and livestock. Therefore, the amount of phosphorus added in the feed must be considered during the feeding process. Phytase is a monomeric protein discovered in corn bran by Suzuki et al. in 1907, and the presence of phytase has been successively detected in various animals, plants and microorganisms. The optimum pH range of plant-derived...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/096C12N11/082C12N11/089C12N11/087C12N11/093C12N11/02C12N9/16C12R1/84
CPCC12N11/08C12N11/02C12N9/16
Inventor 应汉杰丁赛陈勇王芳娟张德力俞莹梁偲策
Owner NANJING UNIV OF TECH
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