A kind of industrial purification method of fiber grade methanol protein

A methanol protein and purification method technology, applied in the field of industrial purification of fiber-grade methanol protein, can solve the problems of low fragmentation rate, difficulty in methanol protein purification, easy inactivation, etc., and achieve the effect of high yield

Active Publication Date: 2021-01-15
义马煤业集团煤生化高科技工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In view of the above situation, in order to overcome the defects of the prior art, the purpose of the present invention is to provide an industrial purification method of fiber-grade methanol protein, which can effectively solve the problem that the cell wall of Pichia pastoris is difficult to break, the fragmentation rate is low, easy to inactivate, and the methanol protein purification is difficult. The problem

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1) Separation of fermentation broth and washing of bacteria

[0025] A. Add 300L of desalted water to 300kg of methanolic protein fermentation broth to dilute, mix, and separate in a high-speed disc centrifuge to obtain 300kg of heavy phase A;

[0026] B. Add 300L of desalted water to 300kg of heavy phase A to dilute, mix evenly, and then enter the high-speed disc centrifuge for centrifugation to obtain 300kg of heavy phase B. The mass concentration of heavy phase B is 50%;

[0027] C. The high-speed disc centrifuge mentioned is produced by GEA Westfalia Separator Co., Ltd. (GEA), the model is HFC 15-01-177, the speed is 11500rpm, the processing capacity is 2t / h, and the use of 1.5mm Nozzle high-speed disc centrifuge;

[0028] 2), cell lysate

[0029] The method is: take 4.5kg of sodium hydroxide and add 9L of desalinated water to prepare a sodium hydroxide solution; take 15L of desalted water, add 1.8kg of sodium lauryl sulfate, stir while heating, heat to 60°C, and ...

Embodiment 2

[0039] 1) Separation of fermentation broth and washing of bacteria

[0040] A. Add desalinated water equal to the weight of the protein methanol fermentation broth to dilute, mix, and separate in a high-speed disc centrifuge to obtain heavy phase A;

[0041] B. Add desalinated water equal to the weight of heavy phase A to the heavy phase A to dilute, mix well, and enter the high-speed disc centrifuge again for centrifugation to obtain heavy phase B. The mass concentration of heavy phase B is 50%;

[0042] C. The high-speed disc centrifuge mentioned is produced by GEA Westfalia Separator Co., Ltd. (GEA), model: HFC 15-01-177, nozzle is 0.5mm nozzle, speed 11500rpm, processing capacity 0.5 t / h high-speed disc centrifuge;

[0043] 2), cell lysate

[0044] The method is, take 1.5% of the total weight of heavy phase B of sodium hydroxide and add desalinated water twice the weight of sodium hydroxide to prepare a sodium hydroxide solution; take 5% of the total weight of heavy phas...

Embodiment 3

[0053] 1) Separation of fermentation broth and washing of bacteria

[0054] A. Add desalinated water equal to the weight of the protein methanol fermentation broth to dilute, mix, and separate in a high-speed disc centrifuge to obtain heavy phase A;

[0055] B. Add desalinated water equal to the weight of heavy phase A to the heavy phase A to dilute, mix well, and enter the high-speed disc centrifuge again for centrifugation to obtain heavy phase B. The mass concentration of heavy phase B is 50%;

[0056] 2), cell lysate

[0057] The method is, take 1.5% of the total weight of heavy phase B of sodium hydroxide and add desalinated water twice the weight of sodium hydroxide to prepare a sodium hydroxide solution; take 5% of the total weight of heavy phase B of desalted water, add the total weight of heavy phase B 0.6% sodium lauryl sulfate, stir while heating, heat to 60 ° C, keep stirring for 10 minutes, until the sodium lauryl sulfate is completely dissolved, the solution is ...

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Abstract

The invention relates to an industrial purification method for fiber-grade methanol proteins and effectively solves the problems of difficult wall breaking of pichia pastoris cells, low breaking rate,easy deactivation and difficult methanol protein purification. The industrial purification method comprises the following steps: performing centrifugal separation on fermentation liquid twice to obtain a heavy phase B; preparing a lysate, adding the lysate in the heavy phase B and adding desalting water to obtain a heavy phase C with a mass concentration of 30 percent; performing high-pressure homogenization to obtain a cell breaking liquid; cooling, regulating a pH value, centrifuging with a centrifugal machine and collecting liquid supernatant to obtain a protein solution; regulating a pH value, performing static precipitation, separating a precipitate with the centrifugal machine and collecting to obtain heavy-phase proteins D; adding desalting water, mixing and centrifuging to obtaineluted re-phase proteins E; regulating pH to obtain a protein alkali solution; drying to obtain the fiber-grade methanol proteins. The industrial purification method provided by the invention has thebenefits that the efficient wall breaking of the pichia pastoris cells as well as the isolation and the purification of the proteins with specific molecular weights are realized, the yield is high, the protein molecular weight distribution is concentrated, the content of crude proteins is above 90 percent, and the requirements for wet spinning of protein fibers can be met.

Description

technical field [0001] The invention relates to the field of protein purification, in particular to an industrial purification method of fiber grade methanol protein. Background technique [0002] Methanol protein is a kind of single-cell protein produced by liquid ventilation fermentation and separation of yeast cells from the fermentation broth with methanol and inorganic salts as the main raw materials. Methanol protein has a crude protein content of more than 50%, balanced amino acids, and high nutritional value, but it cannot be used in feed, food and other industries at present. To develop downstream products of methanol protein and seek new application fields of methanol protein is to solve the problem of methanol overcapacity. It is of far-reaching significance to reduce environmental pollution, develop the national economy, protect the ecological environment and promote the healthy development of related industries. [0003] Methanol protein is a microbial single-c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/16C12N1/06C12R1/84
CPCC12N1/063C12N1/16
Inventor 苟万晓张敬东赵利军杨刚范延超任晓辉胡元森卫红伟白冠章张芳吕昂谢军侯龙龙曹淼郝景梅
Owner 义马煤业集团煤生化高科技工程有限公司
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