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Limulus plasma protein hydrolysis method

A plasma protein and plasma technology, applied in the field of bioengineering, can solve the problems of discarding and wasting Limulus plasma, and achieve the effects of no solvent pollution, less investment and safe operation

Active Publication Date: 2016-12-07
GUANGDONG OCEAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research on the utilization of horseshoe crab blood in my country is mainly to extract the amebocytes in the horseshoe crab blood, and then isolate the small molecular active peptide with antibacterial and anti-endotoxin activity from the amebocytes and produce the limulus reagent. Peptides are used as lead compounds to develop new antibacterial, antiviral, and antitumor drugs. However, the remaining Limulus plasma after the separation of amebocytes from Limulus blood exceeds the human consumption standard, and the Limulus plasma is generally discarded as waste, which has caused great harm. waste
Up to now, no direct research on Limulus plasma proteolytic peptides has been reported at home and abroad

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A method for hydrolysis of Limulus plasma protein, the method comprises the steps:

[0023] S1: The limulus blood is kept in a water bath at a constant temperature of 90 °C for 3 hours, then the limulus blood is filtered and washed with distilled water for several times for use;

[0024] S2: Use trypsin to hydrolyze the limulus blood obtained in step S1, and then centrifuge the limulus blood hydrolysate to obtain a supernatant, which is freeze-dried to obtain a limulus plasma protein hydrolyzed peptide powder.

[0025] The optimal enzymolysis conditions of this example: enzyme concentration 3%, substrate concentration 4%, pH 8.0, hydrolysis time 3h, and hydrolysis temperature 50°C, the hydrolysis rate at this time is 78.29%, and the copper ion content in the hydrolyzed peptide is 44.10 ug / mL.

Embodiment 2

[0027] A method for hydrolysis of Limulus plasma protein, the method comprises the steps:

[0028] S1: The limulus blood is kept in a water bath at a constant temperature of 70 °C for 4 hours, then the limulus blood is filtered and washed with distilled water for several times for use;

[0029] S2: Use trypsin to hydrolyze the limulus blood obtained in step S1, and then centrifuge the limulus blood hydrolysate to obtain a supernatant, which is freeze-dried to obtain a limulus plasma protein hydrolyzed peptide powder.

[0030] The enzymolysis conditions of the present embodiment: enzyme concentration 3%, substrate concentration 5%, pH 8.0, hydrolysis time 2h and hydrolysis temperature 50°C, the hydrolysis rate at this time is 66.09%, and the copper ion content in the hydrolyzed peptide is 47.60ug / mL.

Embodiment 3

[0032] A method for hydrolysis of Limulus plasma protein, the method comprises the steps:

[0033] S1: The limulus blood is kept in a water bath at a constant temperature of 90 °C for 3 hours, then the limulus blood is filtered and washed with distilled water for several times for use;

[0034] S2: using trypsin to hydrolyze the limulus blood obtained in step S1, and then centrifuging the limulus blood hydrolysate to obtain a supernatant, which is freeze-dried to obtain a limulus plasma protein hydrolyzed peptide powder.

[0035] The optimal enzymatic hydrolysis conditions of this example: the enzyme concentration is 3%, the substrate concentration is 5%, the pH is 8.0, the hydrolysis time is 2h, and the hydrolysis temperature is 50°C. The hydrolysis rate at this time is 57.10%, and the copper ion content in the hydrolyzed peptide is 44.10ug / mL.

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Abstract

The invention relates to a limulus plasma protein hydrolysis method. The limulus plasma protein hydrolysis method comprises the following steps of (1) performing constant-temperature water bath on limulus plasma for 2-4h, sucking and filtering limulus plasma, and washing by water for multiple times for standby use; S2, performing proteolysis on limulus blood obtained in step 1, centrifuging limulus plasma hydrolysate, separating to obtain a supernatant, freezing, and drying, so as to obtain limulus plasma protein hydrolysis peptide powder, wherein the water bath temperature in step S1 is 70-100 DEG C, and the pH (potential of hydrogen) value of a hydrolysis system in step S2 is 7-9. The limulus plasma protein hydrolysis method has the advantages that by utilizing protease to perform enzymatic hydrolysis at most suitable temperature and pH condition, the reaction is mild, the degree of hydrolysis can be controlled, and the nutritional value of amino acid is remained; the experiment equipment is simple, the operation is safe, the pollution by solvent is avoided, and the investment is less.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a method for hydrolysis of horseshoe crab plasma protein. Background technique [0002] Protein is one of the essential nutrients for the human body, but different proteins have different digestion and absorption rates in the human body due to their different amino acid composition and spatial structure. Hydrolysis of proteins into polypeptides can greatly improve their absorption rate. The main methods of protein hydrolysis include acid hydrolysis, alkaline hydrolysis, enzymatic hydrolysis, microbial degradation and so on. The chemical hydrolysis method is simple, inexpensive, and has high hydrolysis efficiency, but it has no specificity for peptide chain breakage, the degree of hydrolysis is not easy to control, and the reaction conditions are severe. The microbial degradation method is not easy to control the degradation products and product types in the degradation process, and...

Claims

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Application Information

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IPC IPC(8): C12P21/06
CPCC12P21/06
Inventor 邓春梅杜建喜康信煌何兰珍温燕梅
Owner GUANGDONG OCEAN UNIVERSITY
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