Preparation method of high-molecular-weight squid skin collagen

A high molecular weight, skin collagen technology, applied in the field of protein materials in biotechnology, can solve the problems of low utilization level of squid skin collagen and the difference in molecular weight of squid skin collagen, achieving low cost, good mechanical properties, and maintaining activity. Effect

Active Publication Date: 2012-09-19
ZHEJIANG MARINE DEV RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The purpose of the present invention is to overcome the huge difference in molecular weight between the squid skin collagens prepared by the current technology. If it is used in biomedical materials, it will cause the shortcoming that the utilization level of squid skin collagens is not high. Preparation method of high molecular weight squid skin collagen for biomedical materials with high tensile strength

Method used

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  • Preparation method of high-molecular-weight squid skin collagen

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Embodiment 1

[0043] Take 0.4 Kg of squid skin, wash it with clean water after thawing and mash it, add 30 ml of 0.1mol / L NaOH solution to each gram of squid skin, add 12L NaOH solution, soak at 4°C for 24 hours, then add 30 ml per gram of squid skin Add 12L of n-butanol solution to 10% n-butanol solution by volume, stir for 16 hours, and drain. Add 10ml of distilled water to each gram of squid skin and add 4L of distilled water to the squid skin. 0.4g of Bacillus subtilis neutral protease at 0.1% by weight, hydrolyze at 4°C for 6 h, then adjust the pH to 1 with hydrochloric acid, add 1.2g of pepsin at 0.3% by weight of squid skin, and continue to enzymolyze at 4°C After 16 hours, select an ultrafiltration membrane with a molecular cut-off of 100kd to carry out centrifugal ultrafiltration on the enzymolysis solution, adjust the pH of the filtrate to 4 with ammonia water, and then add ammonium sulfate to the filtrate until the concentration is 2 mol / L, so that the squid skin Collagen is salt...

Embodiment 2

[0046] Take 0.4Kg squid skin, wash it with clean water after thawing and mash it, add 40 ml 0.3 mol / L NaOH solution to each gram of squid skin, add 16 L NaOH solution, soak at 5°C for 28 hours, then add 40 ml per gram of squid skin Add 16L of n-butanol solution to 10% n-butanol solution in ml volume percent, stir for 18 h and drain, add 15 ml of distilled water per gram of squid skin to add 6 L of distilled water to the dried squid skin, add as 0.8g of Bacillus subtilisin neutral protease with 0.2% weight of squid skin, enzymatic hydrolysis at 7°C for 8 hours, then adjust the pH to 1.2 with hydrochloric acid, then add 1.6g of pepsin with 0.4% weight of squid skin, continue at 7°C After 18 hours of enzymolysis, the enzymolyzed solution was centrifuged and ultrafiltered with an ultrafiltration membrane with a molecular cut-off of 100kd, and the pH of the filtrate was adjusted to 3 with ammonia water, and then sodium phosphate was added to the filtrate until the concentration was ...

Embodiment 3

[0049] Take 0.4 Kg squid skin, wash it with clean water after thawing and mash it, add 50 ml of 0.5 mol / L NaOH solution per gram of squid skin, add 20 L NaOH solution, soak at 10°C for 30 h, and then add per gram of squid skin Add 20 L of n-butanol solution to 50 ml of 10% n-butanol solution by volume, stir for 24 h and drain, add 20 ml of distilled water per gram of squid skin to the drained squid skin, add 8 L of distilled water, Add 1.2 g of Bacillus subtilisin neutral protease with 0.3% weight of squid skin, enzymatically hydrolyze at 10°C for 10 h, adjust the pH to 1.5 with hydrochloric acid, add 2.0 g of pepsin with 0.5% weight of squid skin, and continue at 10°C Enzymolysis was performed for 20 h, and the enzymolysis solution was centrifugally ultrafiltered with an ultrafiltration membrane with a molecular cut-off of 100 kd. The pH of the filtrate was adjusted to 5 with ammonia water, and then sodium chloride was added to the filtrate until the concentration was 2.5 mol / ...

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Abstract

The invention discloses a preparation method of high-molecular-weight squid skin collagen, comprising the steps of after thawing the squid skin, using clear water to clean the squid skin and mashing; adding NaOH solution to soak the squid skin; adding n-butyl alcohol solution, and stirring; using distilled water to wash the squid skin until that pH is neutral; drying the squid skin; adding distilled water to the dried squid skin, stirring uniformly; adding protease to perform enzymatic hydrolysis for the squid skin; using ultrafiltration membrane with molecular weight cutoff of 100kd to filter the enzymolysis liquid; collecting filtering substances on the ultrafiltration membrane; and freeze-drying to obtain the high-molecular-weight squid skin collagen. The biomedicine material prepared by the high-molecular-weight squid skin collagen obtained by the method has good mechanical property; the method has simple process step, can recover low-molecular-weight collagen, improve whole utilization level of the squid skin, can improve utilization value of the squid skin and can reduce environment pollution level.

Description

[0001] technical field [0002] The invention relates to the field of protein materials in biotechnology, in particular to a method for preparing high-molecular squid skin collagen. [0003] Background technique [0004] Collagen is an important class of structural proteins that widely exist in supporting tissues such as bones, epidermis, and tendons of multicellular animals. Due to its weak antigenicity, good tissue compatibility, gel type, thickening, emulsifying Collagen and its hydrolyzate have a wide range of uses and good application prospects in the fields of medical care, beauty, food and daily chemical industry, while the current commercial collagen is usually derived from slaughtered animals containing connective tissue In recent years, the emergence of mad cow disease, foot-and-mouth disease, bird flu and other diseases has greatly restricted the application of collagen derived from terrestrial animals, while marine animals are rich in collagen and have great pot...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/36C07K1/34C07K1/30
Inventor 付万冬钟明杰廖妙飞郑斌杨会成王维伦
Owner ZHEJIANG MARINE DEV RES INST
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