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High f value oligopeptide and preparation method thereof

A technology of oligopeptide and pH value, applied in the field of high F value oligopeptide and its preparation, can solve the problems of large-scale production difficulties, difficult process amplification, and small loading amount

Active Publication Date: 2020-07-07
湖北跃莱生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to research reports at home and abroad, Sephadex G~15 or Bio~Gel P~2 gel chromatography is generally used for separation and purification, but because the amount of sample loaded is small and the process is not easy to scale up, it brings difficulties to large-scale production

Method used

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  • High f value oligopeptide and preparation method thereof
  • High f value oligopeptide and preparation method thereof
  • High f value oligopeptide and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1 Preparation of yeast protein aqueous solution

[0054] Distilled water is added to the active dry yeast to form a yeast protein solution with multiple groups of certain solid content. The concentration can be selected from 160 to 210 g / L, and the solution is ultrasonically broken. The ultrasonic power is 300W, 350W, 400W or 450W, the ultrasonic time is 20min, and the time interval is 2s / 2s.

[0055] Taking the soluble protein in the supernatant after crushing as an index, with the strengthening of ultrasonic power, the degree of yeast fragmentation first increases and then tends to be flat. When the power reaches 400W, the degree of change in protein yield tends to be stable, so consider the issue of energy consumption. , choose 400W ultrasonic power for ultrasonic crushing.

[0056] Then select the ultrasonic power to be 400W, configure multiple groups of yeast protein solutions according to the above method, and ultrasonicate at 400W for 10min, 15min, 20...

Embodiment 2

[0058] Example 2 Preparation of Yeast Extract

[0059] Configure the yeast protein solution according to the same steps as in Example 1, ultrasonicate the yeast protein solution at 400W for 20 minutes, and then use neutral protease, pepsin, alkaline protease and trypsin to enzymatically hydrolyze the solution, respectively set at a more suitable temperature and pH, the reaction time is 10h, and the amount of enzyme added is 500U·g -1 .

[0060] The enzymatic hydrolysis conditions are as follows:

[0061] Neutral protease: 40-50°C, pH=6.0-7.0.

[0062] Pepsin: 35-45°C, pH=1.5-2.5.

[0063] Alkaline protease: 45-55°C, pH=7.0-9.0.

[0064] Trypsin: 35-45°C, pH=7.5-8.5.

[0065] The result is as figure 2 shown by figure 2 It can be seen that the degree of hydrolysis of yeast protein by alkaline protease is the highest, followed by neutral protease, and the hydrolysis effect of pepsin is the lowest. At the same time, the protein recovery rate was positively correlated wit...

Embodiment 3

[0066] Example 3 Nanofiltration Concentration of Yeast Extract

[0067] On the basis of Example 2 (alkaline protease is used as the enzyme for active dry yeast extraction), the yeast extract was obtained, then the enzyme was inactivated in a boiling water bath for 15 minutes, and the supernatant was taken after centrifugation at 13000r / min for 15 minutes. Then get the supernatant and carry out the nanofiltration concentration operation. The nanofiltration condition is to adopt a PES roll-type membrane with a molecular weight cut-off of 500Da, the operating pressure is 2-4MPa, the initial pH of the yeast extract is 6.5-7.5, and the concentration ratio (volume of the cut-off liquid The ratio to the volume of the stock solution (1 / 1.5~1 / 3) is 1.0, 1.5, 2.0, 2.5, 3.0 times (concentration of 3.0 times means that the volume ratio of the retentate to the stock solution is 1 / 3, and other concentration ratios have similar meanings here ).

[0068] The result is as image 3 As shown, ...

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Abstract

Provided is a preparation method for a high-F ratio oligopeptide, comprising the following steps: adding an alkaline protease to a yeast protein solution, and performing enzymolysis under conditions of pH 7.0-9.0 at 45 to 55 °C to obtain a yeast extract; concentrating the yeast extract to remove molecules having a molecular weight of 500 Da or less to obtain a concentrate; adding alpha-chymotrypsin to the concentrate, and performing enzymolysis under the conditions of pH 6.5-8.5 at 35 °C to 45 °C for 2 to 8 h; and inactivating enzymes, adding carboxypeptidase A to the enzymatic hydrolysate, performing enzymolysis under conditions of pH 6.5-7.5 at 35 °C to 45 °C for 4 to 8h, inactivating said enzyme, and using activated carbon to adsorb and remove impurities, so as to obtain the high F-ratio oligopeptide.

Description

technical field [0001] The invention relates to the technical field of enzyme preparations, in particular to a high F value oligopeptide and a preparation method thereof. Background technique [0002] High F value oligopeptide is a mixed small peptide mixture composed of 2 to 9 amino acid residues. F value refers to the branched chain amino acids (leucine, isoleucine and valine, BCAA for short) and aromatic The molar ratio of amino acids (phenylalanine, tyrosine, referred to as AAA) is named in commemoration of the "pseudo-neurotransmitter hypothesis" proposed by the famous German scholar Fischer in the 1970s. The F value of high F value oligopeptide should be greater than 20. Oligopeptides are protein precursors composed of 2 to 9 amino acids, or protein degradation products composed of 2 to 9 amino acids, and can also be called small peptides, short peptides, etc. In actual production, high F value oligopeptides can be widely used in drugs for the treatment of liver dise...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/06
CPCC12P21/06
Inventor 田亚平吴警涛李婷婷周楠迪
Owner 湖北跃莱生物工程有限公司
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