Method for preparing active polypeptides from deep-sea fish meat

A technology of active peptides and deep-sea fish, applied in the direction of microorganism-based methods, biochemical equipment and methods, peptide sources, etc., can solve the problems of poor competitiveness, low added value of products, single product structure, etc., and achieve the goal of improving stability Effect

Inactive Publication Date: 2018-01-19
兰溪市哥特生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The total amount of marine biological protein resources in China ranks among the best in the world. However, in my country's current protein aquatic products, except for a part of direct consumption, most of them are made into food thr

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] The method for preparing an active polypeptide by utilizing deep-sea fish meat comprises the following steps:

[0022] 1) Degreasing: cut deep-sea fish meat into small pieces, add 2~4‰ chitosan immobilized lipase, add 8~15 times the volume of distilled water, and enzymolyze at a constant temperature of 25~50℃ for 0.8~2.5h, and enzymolyze After the end, the enzyme is inactivated, and the defatted deep-sea fish meat is obtained by filtration. The preparation steps of chitosan-immobilized lipase are as follows: use dry chitosan powder, soak overnight with 60-80mmol / L phosphate buffer solution of pH 6.0-6.8, then add 0.7-1.2% chitosan powder mass 1-ethyl-3-(3-methylamino)propylcarbodiimide was activated for 0.5~0.9h, then chitosan powder 1~2 times the volume of glutaraldehyde coupling reaction was added for 1~1.8h, with Wash with distilled water for 5-8 times, add 0.7-2.0mg / mL lipase solution and 0.1-0.3‰ active short peptide, stir slowly in an ice bath for 0.5-1.5h, then ...

Embodiment 2

[0030] The most preferred method for preparing active polypeptides from deep-sea fish meat comprises the following steps:

[0031] 1) Degreasing: cut deep-sea fish meat into small pieces, add 3.2‰ chitosan immobilized lipase, add 10 times the volume of distilled water, and enzymolyze at a constant temperature of 30°C for 1.4 hours. Deep-sea fish meat defatted. The preparation steps of chitosan-immobilized lipase are: use dry chitosan powder, soak overnight with 65mmol / L phosphate buffer solution of pH 6.3, then add 1.0% 1-ethyl-3 -(3-Methylamino)propyl carbodiimide activation for 0.7h, then add chitosan powder 1.4 times the volume of glutaraldehyde coupling reaction for 1.3h, wash with distilled water 5 times, add 1.2mg / mL lipase Solution and 0.3‰ active short peptide were slowly stirred in an ice bath for 1.1h, then placed in the refrigerator overnight, rinsed thoroughly with distilled water the next day, filtered and dried to obtain chitosan-immobilized lipase. The amino a...

Embodiment 3

[0039] The method for preparing an active polypeptide by utilizing deep-sea fish meat comprises the following steps:

[0040] 1) Degreasing: Deep-sea fish meat is cut into small pieces, added with 3‰ chitosan immobilized lipase, added 11 times the volume of distilled water, and enzymatically hydrolyzed at a constant temperature of 30°C for 1.2 hours. Deep-sea fish meat defatted. The preparation steps of chitosan-immobilized lipase are: use dry chitosan powder, soak overnight with 70mmol / L phosphate buffer solution of pH6.5, then add 1-ethyl-3 -(3-Methylamino)propyl carbodiimide activation for 0.8h, then add chitosan powder 1.3 times the volume of glutaraldehyde coupling reaction for 1.2h, wash with distilled water 6 times, add 1.3mg / mL lipase Solution and 0.2‰ active short peptide were slowly stirred in an ice bath for 1.0 h, then placed in the refrigerator overnight, rinsed thoroughly with distilled water the next day, filtered and dried to obtain chitosan-immobilized lipase...

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PUM

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Abstract

The invention discloses a method for preparing active polypeptides from deep-sea fish meat, and the method includes degreasing, first deodorization, enzymolysis, second deodorization, ultrafiltration,decolorization, desalting and freeze-drying. 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide is used to activate chitosan, and coupled with glutaraldehyde to form a lipase immobilized carrier, an active short peptide is used to improve the physical and chemical adsorption force between a carrier and lipase, the lipase stability, activity, heat resistance and use frequency are improved; through useof the complementary effect and synergistic effect of thermolysin and chymotrypsin hydrolysis sites, the protein hydrolysis degree and enzymatic hydrolysis speed of minced deep-sea fish meat can be improved, terminal group type can be changed, and the active polypeptides with antitumour activity can be obtained by hydrolysis.

Description

technical field [0001] The invention relates to the field of preparation of biologically active substances, in particular to a method for preparing active polypeptides using deep-sea fish meat. Background technique [0002] Although local cancers can be successfully treated with surgery and radiotherapy, chemotherapy is still the first choice for conventional treatment of advanced or metastatic tumors. Chemotherapeutic drugs have defects such as low selectivity, large adverse reactions, and multidrug resistance, which largely limit their use. [0003] Due to the low content of naturally occurring marine active peptides in organisms and the difficulty of extraction, it is difficult to meet the needs of mass production and supply, and the cost of chemical synthetic peptides is expensive. Marine biological protein resources are an important protein food and an important source of biologically active substances for human beings in the 21st century. The total amount of marine b...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K14/46C07K1/36C07K1/34C12N11/10C12R1/865C12R1/01
Inventor 李健
Owner 兰溪市哥特生物技术有限公司
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