High-F-value oligopeptide and preparation method thereof

An oligopeptide and pH value technology, applied in the field of high F value oligopeptide and its preparation, can solve the problems of less sample loading, difficult process amplification, difficulty in large-scale production, etc., and achieve sufficient sources, low price, and high F value. Effect

Active Publication Date: 2018-11-27
湖北跃莱生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 purifica

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

The invention relates to a preparation method of a high-F-value oligopeptide. The preparation method of the high-F-value oligopeptide comprises the following steps of adding alkaline proteases into ayeast protein solution for emzymolysis at a pH of 7.0-9.0 and a temperature of 45-55 DEG C to obtain a yeast leaching liquor, wherein the adding amount of the alkaline proteases is 400-600 U/g to yeast proteins; concentrating the yeast leaching liquor to removing molecules with the molecular weight lower than 500 Da in the yeast leaching liquor to obtain a concentrated solution; adding alpha-chymotrypsin into the concentrated solution for emzymolysis for 2-8 h at a pH of 6.5-8.5 and a temperature of 35-45 DEG C; performing enzyme deactivation, and then adding carboxy-peptidase A into the enzymatic products for emzymolysis for 4-8 h at a pH of 6.5-7.5 and a temperature of 35-45 DEG C, performing enzyme deactivation and absorbing impurities through activated carbon to obtain the high-F-valueoligopeptide. The preparation method of the high-F-value oligopeptide is simple, low in cost, rich in material resources and controllable during emzymolysis and can prepare the purely natural and absorbable high-F-value 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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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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