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Pichia pastoris with endotoxin removal activity and application of pichia pastoris

A Pichia pastoris and endotoxin technology, applied in the direction of microorganism-based methods, microorganisms, fungi, etc., can solve the problems of endangering the health of livestock and poultry, insufficient attention, and affecting the economic benefits of breeding, etc., and achieve high-efficiency endotoxin detoxification performance , the effect of extensive application value

Inactive Publication Date: 2018-11-16
SHANDONG BIO SUNKEEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Endotoxin is the outer wall component of Gram-negative bacteria and one of the main pathogenic factors. After the bacteria are contaminated in the feed, the endotoxin released can endanger the health of livestock and poultry and affect the economic benefits of breeding
The endotoxin released by Gram bacteria in food can cause human poisoning, but people have not paid enough attention to it

Method used

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  • Pichia pastoris with endotoxin removal activity and application of pichia pastoris
  • Pichia pastoris with endotoxin removal activity and application of pichia pastoris
  • Pichia pastoris with endotoxin removal activity and application of pichia pastoris

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Screening of endotoxin-detoxified strains

[0026] The soil was collected from the factory of Shandong Shengqi Biological Co., Ltd., diluted with sterile water, spread and isolate a series of single colonies, cultured in liquid until the early logarithmic phase, washed with PBS, and made into OD 600 The PBS bacterial suspension equal to 6 is for use, selects Sigma company E. coli The endotoxin derived from O111:B4 was made into a PBS solution with a concentration of 4 mg / ml for use. Mix the PBS bacterial suspension with the endotoxin-removing solution, incubate at 30 degrees, take samples at 10 minutes, 20 minutes, 30 minutes, 40 minutes, and 50 minutes, and centrifuge to get the supernatant for KDO quantitative detection, and select the strain with the greatest detoxification effect. in P. pastoris Sunkeen-66 detoxification kinetic curve. like figure 1 As shown, the endotoxin detoxification characteristics of the Pichia pastoris strains, using the k...

Embodiment 2

[0027] Example 2 Investigation of the detoxification performance of Pichia pastoris Sunkeen-66 cell wall for endotoxin in feed

[0028] Put the culture medium into the fermenter, sterilize it, then inoculate the fermented yeast Pichia pastoris with the preservation number CCTCC NO: M2017723, inoculate it into the fermenter, and add nutrient solution for cultivation; ferment the fermented liquid at 3000- Centrifuge at 6000 rpm to collect yeast sludge, wash with deionized water, and then centrifuge at 3000-6000 rpm to collect Pichia Sunkeen-66 cells, inactivate at 80-95 °C for 30 min, cool down at 55-65 °C, Add a certain amount of protease, hydrolyze for 5-15 hours, centrifuge, collect the heavy phase, and prepare the cell wall fraction for later use;

[0029] Then divide the corn feed into 2 parts, one of which is contaminated with endotoxin (concentration is 1 mg / g), and then divide the 2 parts of corn feed into 2 small parts, of which 1 small part is added with cell wall comp...

Embodiment 3

[0030] Example 3 Investigation of Pichia pastoris Sunkeen-66 fermentation broth for endotoxin detoxification performance in drinking water

[0031] First, by fermenting Sunkeen-66 cells, take the fermentation broth, centrifuge at 6000 rpm for 10 min, and collect the supernatant for later use; then divide the pure water into 2 parts, one of which is contaminated with endotoxin (concentration is 10 ug / g), and one without pollution; and then divide these 2 parts of water into 2 small parts respectively, wherein 1 small part is added with the above-mentioned fermentation liquid supernatant according to the proportion of 20%, and the other 1 small part is added with purified water according to the proportion of 20%, forming a completely random 2 × 2 analysis Due to the design of the analysis, there are 4 types of drinking water that are polluted or not polluted with endotoxin, added or not added with Sunkeen-66 fermentation supernatant. Keep warm at 30 degrees, take a sample and di...

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Abstract

The invention discloses pichia pastoris with endotoxin removal activity and an application of the pichia pastoris in endotoxin removal. The pichia pastoris with endotoxin removal activity is screenedby high-throughput breeding, and the pichia pastoris is preserved in CCTCC (China Center for Type Culture Collection) with the preservation number being CCTCC NO: M2017723. The invention further discloses the application of the pichia pastoris in the endotoxin removal. Whole cells, cell wall components or fermentation liquor of the pichia pastoris are added to endotoxin polluted feed, food or fermentation liquor, and residual level of endotoxin is reduced to 60% or lower. The pichia pastoris can be applied to the endotoxin removal of products such as feed, food, biological products and the like.

Description

Technical field [0001] The invention discloses a Pichia pastoris with endotoxin detoxification activity and its application in endotoxin detoxification. Background technique [0002] Endotoxin is the outer wall component of Gram-negative bacteria and one of the main pathogenic factors. After the bacteria are contaminated in the feed, the endotoxin released can endanger the health of livestock and poultry and affect the economic benefits of breeding. The endotoxin released by Gram bacteria in food can cause human poisoning, but people have not paid enough attention to it. Endotoxin (pyrogen) contamination is common in biological products, so it is of great significance to find microbial strains with endotoxin detoxification activity. [0003] Yeast is a large class of unicellular eukaryotic microorganisms widely used in food brewing, feed additives, biochemical products, and industrial biotechnology. Yeast also has the function of biological detoxification, and has important...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/16A23L5/20C12R1/84
CPCA23L5/28C12N1/165C12R2001/84
Inventor 胡晓清易勇张继祥张亚吴倩尚建丽付传超王栋杜显雨
Owner SHANDONG BIO SUNKEEN
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