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A soybean active peptide additive for cell culture medium

A soybean active peptide and additive technology, applied in animal cells, vertebrate cells, artificial cell constructs, etc., can solve the problems of cell apoptosis, destroy cell membrane structure, change cell shape, etc., achieve low endotoxin content, promote cell Proliferation and high enzymatic hydrolysis efficiency

Active Publication Date: 2018-05-04
CHINA NAT RES INST OF FOOD & FERMENTATION IND CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because soybean active peptides are rich in nutrients, they are easily contaminated by bacterial microorganisms during their processing and storage, and then produce a large amount of bacterial endotoxins. When they are used as cell culture additives, they will change cell morphology, destroy cell membrane structure, and induce cell apoptosis. Death, etc., have adverse effects on the growth of cells cultured in vitro
In addition, because different processing techniques have a great influence on the composition of the prepared soybean active peptide products, and the composition of animal serum is relatively complex, and many components are still unclear, so it is impossible to determine the existence of certain or certain components or Whether the deletion will have adverse effects on the growth of cells, which will cause some difficulties in the application of soybean active peptides in serum-free medium supplements

Method used

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  • A soybean active peptide additive for cell culture medium
  • A soybean active peptide additive for cell culture medium
  • A soybean active peptide additive for cell culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] 1. Enzymatic treatment

[0051] Add 12 times deionized water to soybean protein isolate with protein content ≥ 90%, mix well, heat up to 90°C, keep warm for 15-20 minutes, and when the temperature is lowered to 50°C, adjust the pH value of the protein solution to 6.5- 7.5, add the compound non-specific protease ( Among them, the enzyme amounts of neutral protease PROTAMEX and bromelain are 2000 units and 1500 units respectively), and the first enzymolysis treatment was incubated at 50°C for 4.5 hours to obtain the first enzymolysis solution; the pH value of the first enzymolysis solution was adjusted After it is 7.5-8.5, add a mixture of flavor protease FLAVOURZYME500MG (Novozymes) and carboxypeptidase ACCELERZYME CPG (DSM) to the first enzymolysis solution with an enzyme amount of 2500 units per gram of protein Compound specific proteases (the enzyme amounts of flavor protease FLAVOURZYME500MG and carboxypeptidase ACCELERZYME CPG are 1500 units and 1000 units respecti...

Embodiment 2

[0070] The enzymatic hydrolysis treatment is specifically: add 9 times deionized water to soybean protein isolate with protein content ≥ 90%, mix well, heat up to 90-95°C, keep warm for 55-60 minutes, then cool down to 50-55°C When adjusting the pH value of the protein solution to 7-8, add papain (Nanning Pangbo Biological Engineering Co., Ltd.) and alkaline protease ALKALINE PROTEASE CONCENTRATE (DSM Company) mixed compound non-specific protease (the enzyme amounts of papain and alkaline protease ALKALINE PROTEASE CONCENTRATE are 1000 units and 2000 units respectively), and the first enzymolysis treatment was incubated at 50-55 ° C for 2.5 hours to obtain the first 1. Enzymolysis solution; after adjusting the pH value of the first enzymolysis solution to 8-9, add flavor protease FLAVOURZYME500MG (Novozymes) to the first enzymolysis solution with an enzyme amount of 2000 units per gram of protein Mix the compound specific protease with carboxypeptidase Validase FP Conc. (DSM c...

Embodiment 3

[0073] The enzymatic hydrolysis treatment is specifically as follows: add 10 times deionized water to soybean protein isolate with protein content ≥ 90%, mix evenly, heat up to 95°C, keep warm for 35-40 minutes, and then cool down to 45-50°C, Regulate the pH of the protein solution to be 7-8, and add papain (Nanning Pangbo Bioengineering Co., Ltd.), bromelain (Nanning Pangbo Biological Engineering Co., Ltd.) to the protein solution with an enzyme amount of 4500 units per gram of protein. company) and alkaline protease Novozym37071 (Novozym Company) mixed non-specific protease (in which the enzyme amount of papain, bromelain and alkaline protease are 1000 units, 1500 units and 2000 units respectively), at 45~ Incubate the first enzymatic hydrolysis treatment at 50°C for 3.5 hours to obtain the first enzymatic hydrolysis solution; then add carboxypeptidase Validase FP Conc. (DSM Company) and flavor protease MAXZPRO XF (DSM company) compound specific protease (enzyme amount of ca...

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Abstract

The invention provides a soybean active peptide additive for cell culture medium, in which the oligopeptides with a molecular weight less than 1000 Daltons account for ≥90% of the total protein by mass, and the oligopeptides are at least Including one or more of YE, ERQ, QDPQ, EGGAH, PQGAR, AAGGSN. The soybean active peptide additive of the present invention can be compounded with a variety of basal media for serum-free culture of a variety of animal cells, which not only greatly reduces the cost of cell culture media, but also reduces It can also promote cell proliferation, improve cell viability and enhance the expression of cell products.

Description

technical field [0001] The invention relates to a cell culture medium additive, in particular to a soybean active peptide additive for a cell culture medium. Background technique [0002] Soybean protein isolate is a full-price protein food additive produced from low-temperature desolvated soybean meal. Its protein content is as high as 90%, and there are nearly 20 kinds of amino acids, including essential amino acids for human body. Soy active peptide is a peptide mixture obtained by separating and refining soybean protein through microbial fermentation, acid hydrolysis or enzymatic hydrolysis. It is mainly composed of small molecular peptides, and also contains a small amount of macromolecular peptides, free amino acids, sugars and Inorganic salts and other ingredients. Soybean active peptides are widely used in food, medicine, daily chemical industry and other fields because of their good nutritional properties, health care properties and processing properties. [0003]...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071C12P21/06C07K1/36C07K1/34C07K1/18
Inventor 蔡木易林峰刘艳张海欣谷瑞增鲁军魏颖
Owner CHINA NAT RES INST OF FOOD & FERMENTATION IND CO LTD
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