Method for industrially producing grouper growth hormone recombination gene protein

A growth hormone and recombinant gene technology, applied in the field of industrial production of grouper growth hormone recombinant gene protein, can solve the problems of low yield, long time consumption, high cost, etc., achieve high yield, reduce production cost, and promote healthy growth

Inactive Publication Date: 2013-04-03
GUANGDONG HAID ANIMAL HUSBANDRY & VETERINARY RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The existing technology is to extract the growth hormone from the grouper, introduce the recombinant expression vector, transfer it into Pichia pastoris, and then manually cultivate the recombinant Pichia pastoris containing the growth hormone of the grouper in the laboratory Fermentation, the fermentation time is as long as 5 days, it takes a long time, the output is low, and the cost is high, so it cannot be applied and promoted in actual production

Method used

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  • Method for industrially producing grouper growth hormone recombination gene protein
  • Method for industrially producing grouper growth hormone recombination gene protein
  • Method for industrially producing grouper growth hormone recombination gene protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Obtain grouper growth hormone recombinant protein according to the above steps, using a 50L self-controlled tank, wherein the fermentation conditions are controlled as temperature 28°C, pH 5.5, dissolved oxygen 30%, and fermentation time 90h; the fermentation medium in the shake flask culture The volume is 30L, and the inoculum size is 1.5L; the methanol by volume accounts for 0.55% of the fermentation broth volume.

Embodiment 2

[0045] According to the above steps, the recombinant protein of grouper growth hormone was obtained, using a 50L self-controlled tank, wherein the fermentation conditions were controlled as temperature 29.5°C, pH 6.0, dissolved oxygen 40%, and fermentation time 80h; the fermentation medium in the shake flask culture The volume of the methanol is 30L, and the inoculum size is 1.8L; the volume of the methanol accounts for 0.7% of the fermentation broth volume.

Embodiment 3

[0047] According to the above steps, the recombinant protein of grouper growth hormone was obtained, using a 100L self-controlled tank, wherein the fermentation conditions were controlled as temperature 31.5°C, pH 6.5, dissolved oxygen 40%, and fermentation time 90h; the fermentation medium in the shake flask culture The volume of the methanol is 50L, and the inoculum size is 3.0L; the volume of the methanol accounts for 0.55% of the fermentation broth volume.

[0048] Test results:

[0049] The wet weight of the strains in the fermentation broth was detected, and the protein expression of the target protein in Examples 1-3 was detected by Western Blot and SDS-PAGE polyacrylamide gel electrophoresis. The results are shown in Table 1.

[0050] Table 1: Wet weight and gGH content of strains

[0051] group 1 2 3 Wet weight of bacteria ≤20.75% 59-59.75% ≤30.15% gGH (g / L) ≤0.5 1.5-2.0 ≤0.75

[0052] Qualitative and quantitative detection of reco...

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Abstract

The invention discloses a method for industrially producing grouper growth hormone recombination gene protein. The method comprises the steps of constructing a cloning vector pGEMT-gGH, constructing a yeast expression vector pPICZaA-gGH, acquiring grouper growth hormone recombination gene-containing pichia pastoris, conducting fermentation in self-control tank, conducting aftertreatment and the like. With the method for industrially producing grouper growth hormone recombination gene protein, the industrial production of the grouper growth hormone recombination gene protein can be realized, the production cost can be greatly lowered, and high-output and stable grouper growth hormone recombination gene protein products can be obtained, thereby providing practical products for truly realizing large-scale application in grouper cultivation production, and prompting the health growth of the groupers.

Description

[0001] technical field [0002] The invention relates to the field of bioengineering fermentation, in particular to a method for industrialized production of grouper growth hormone recombinant gene protein, which uses an automatic control tank to produce grouper growth hormone recombinant protein. [0003] Background technique [0004] Growth hormone (GH) is the most important endocrine hormone for fish growth and development, and plays an important role in regulating the growth and development of fish. Grouper growth hormone (gGH) is a protein polypeptide with a molecular weight of 22kD synthesized and secreted by the pituitary gland. Genetic engineering technology can be used to express fish growth hormone protein in engineering bacteria to obtain a large number of cheap growth hormone protein products , Through a special way, such as mixing gGH in the feed, the cultured fish can directly obtain the growth hormone from the outside, thereby increasing the growth rate and r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/81C12R1/84
Inventor 石和荣张勇赵会宏潘忠超
Owner GUANGDONG HAID ANIMAL HUSBANDRY & VETERINARY RES INST
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