Method for extracting earthworm protein and lumbrokinase from earthworms

An extraction method, lumbrokinase technology, is applied in the field of radio frequency technology-assisted lumbrokinase extraction and earthworm protein enzymatic hydrolysis, which can solve the problems of enzyme activity loss, small processing capacity, unsuitability for industrial production, etc., to prevent protease contamination and deterioration, and product quality Excellent effect

Inactive Publication Date: 2018-05-18
天津百利食品股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purification method of lumbrokinase is usually salting out of a single species, and then obtained from different strains of earthworms through various chromatography, HPLC, FPLC and even affinity chromatography or DEAE ion exchange chromatography, molecular sieve, gel electrophoresis and other means. A variety of simple components of plasmin, but the operation is complicated, the process is long, the processing volume is small, and the enzyme activity is greatly lost, and it is not suitable for industrial production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] (1) Take fresh plump live earthworms, wash them several times with water, and discharge the dirt in their viscera as much as possible, then remove them and use filter paper to absorb excess water as much as possible, and place them in a radio frequency system for 30 minutes. The radio frequency is 100MHz and the temperature 37°C;

[0028] (2) Add 0.1mol / L pH7.8 phosphate buffer solution and 0.2% sodium chloride aqueous solution pre-cooled at 4°C to homogenize for 5 minutes in a homogenizer, leave to stand in a refrigerator at 4°C for 4 hours, and then centrifuge. Discard the precipitate to obtain the supernatant;

[0029] (3) Take the supernatant and add solid ammonium sulfate to reach 35% saturation, centrifuge at 4000r / min for 10min, collect the supernatant, then add solid ammonium sulfate to reach 70% saturation, let stand overnight, centrifuge at 15000r / min, collect Precipitate, then dissolve the precipitate with 0.1mol / L phosphate buffer, and filter to obtain a cr...

Embodiment 2

[0032] (1) Take fresh plump live earthworms, wash them several times with water, and discharge the dirt in their viscera as much as possible, then remove them and use filter paper to absorb excess water as much as possible, and place them in a radio frequency system for 30 minutes. The radio frequency is 150MHz and the temperature 40°C;

[0033] (2) Add 0.1mol / L pH7.8 phosphate buffer solution and 0.2% sodium citrate aqueous solution pre-cooled at 4°C, homogenize in a homogenizer for 5 minutes, let stand in a refrigerator at 4°C for 4 hours, and then centrifuge at high speed, Discard the precipitate to obtain the supernatant;

[0034] (3) Take the supernatant and add solid ammonium sulfate to reach 35% saturation, centrifuge at 4000r / min for 10min, collect the supernatant, then add solid ammonium sulfate to reach 70% saturation, let stand overnight, centrifuge at 20000r / min, collect Precipitate, then dissolve the precipitate with 0.1mol / L phosphate buffer, and filter to obtai...

Embodiment 3

[0039] (1) Take fresh plump live earthworms, wash them several times with water, and discharge the dirt in their internal organs as much as possible, then remove them and use filter paper to absorb excess water as much as possible, and place them in a radio frequency system for 30 minutes. The radio frequency is 200MHz and the temperature 50°C;

[0040] (2) Add 0.1mol / L pH7.8 phosphate buffer and 0.2% disodium hydrogen phosphate aqueous solution which were pre-cooled at 4°C, homogenize in a homogenizer for 5 minutes, let stand in a refrigerator at 4°C for 4 hours, and then Centrifuge, discard the precipitate, and obtain the supernatant;

[0041] (3) Take the supernatant and add solid ammonium sulfate to reach 35% saturation, centrifuge at 4000r / min for 10min, collect the supernatant, then add solid ammonium sulfate to reach 70% saturation, let stand overnight, centrifuge at 25000r / min, collect Precipitate, then dissolve the precipitate with 0.1mol / L phosphate buffer, and filt...

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PUM

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Abstract

The invention relates to a method for extracting earthworm protein and lumbrokinase from earthworms. The method is characterized by utilizing an approach to combining chromatographic columns with ultrafiltration for extracting the lumbrokinase and earthworm protein from the earthworms. According to the method, a radio-frequency technique is utilized to achieve a protein extraction sterility mechanism, the appropriate temperature for occurrence of enzyme activity is controlled, and the method is safe, sanitary and free of pollution; by means of the differential control over the earthworm protein, extraction and separation of the earthworm protein are achieved, and batched production of the earthworm protein and lumbrokinase is facilitated.

Description

technical field [0001] The invention relates to a method for extracting worm protein and lumbrokinase from earthworms, in particular to a method for lumbrokinase extraction and worm protein enzymatic hydrolysis assisted by radio frequency technology. Background technique [0002] Earthworms, also known as earthworms, are rich in protein, amino acids, trace elements, enzymes, hormones and other functional ingredients. The dry protein content of earthworms reaches 70%. They are raw materials for the preparation of medicines, cosmetics, health foods, etc., and have great economic value. Lumbrokinase is a serine proteolytic enzyme with kinase and plasmin activity. It can be used as a drug for treating thrombotic diseases. It is a second-class new drug approved by the state for the prevention and treatment of cardiovascular and cerebrovascular diseases. One of the soluble drugs. [0003] Radio frequency (referred to as RF) is a high-frequency AC electromagnetic wave with a frequ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/76C12N9/64C12P21/06C07K1/36C07K1/34C07K1/30
CPCC07K1/303C07K1/34C07K1/36C12N9/6405C12N9/6408C12P21/06C12Y304/21004
Inventor 赵金梁赵发
Owner 天津百利食品股份有限公司
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