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Sericin peptide with alpha-glycosidase inhibiting activity as well as preparation and application thereof

A technology of glycosidase inhibition and sericin peptide, which is applied in application, medical preparations containing active ingredients, peptides, etc., can solve problems such as liver damage, and achieve the effect of lowering postprandial blood sugar and inhibiting the activity of α-glucosidase.

Pending Publication Date: 2017-09-01
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, both acarbose and voglibose can cause gastrointestinal reactions such as abdominal distension and exhaust to a certain extent, and long-term use can cause liver damage

Method used

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  • Sericin peptide with alpha-glycosidase inhibiting activity as well as preparation and application thereof
  • Sericin peptide with alpha-glycosidase inhibiting activity as well as preparation and application thereof
  • Sericin peptide with alpha-glycosidase inhibiting activity as well as preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Preparation of a sericin peptide with α-glucosidase inhibitory activity:

[0047] Wherein, the sericin peptide having α-glucosidase inhibitory activity has the amino acid sequence shown in SEQ ID No:1. Its preparation steps are as follows:

[0048] (1) Weigh n equivalents of resin into the reactor, add DCM to swell for half an hour, then remove the DCM, add 2n equivalents of the first amino acid in the sequence, add 2n equivalents of DIEA, appropriate amount of DMF, DCM (appropriate amount means It is advisable to fully agitate the resin), nitrogen bubbling reaction for 60min. Then add about 5n equivalent of methanol, react for half an hour, remove the reaction solution, and wash with DMF and MEOH;

[0049](2) Add the second amino acid in the sequence (also 2n equivalents), 2n equivalents of HBTU and DIEA to the reactor, react with nitrogen gas bubbling for half an hour, wash off the liquid, detect ninhydrin, and then seal with pyridine and acetic anhydride end. Fin...

Embodiment 2

[0055] Elution and purity determination of synthetic sericin peptide:

[0056] Take a little crude peptide that has been completely synthesized, and dissolve the sample with 10% acetonitrile solution containing 0.1% (v / v) TFA. The gradient elution of phase B from 15% to 48% was carried out as the eluent of 0.1% TFA aqueous solution and 0.1% TFA acetonitrile solution. Such as figure 1 As shown, the sericin peptide synthesized by the present invention has an elution peak with a molecular size of 1379.36Da, and its retention time is 7.752min.

[0057] Take the eluted main peak sample for mass spectrometry, and determine the sample purity according to the required sample molecular weight 1379.36 and the relative area content of the main peak, such as figure 2 As shown, the purity of the synthesized sericin peptide synthesized by the present invention can reach 99.11% through purification.

[0058] When the purity of the peptide reaches above 99%, the secondary preparation and ...

Embodiment 3

[0060] Synthetic sericin peptide inhibits α-glucosidase activity assay:

[0061] Reagents: α-glucosidase (EC 3.2.1.20, 19.3units / mg) was purchased from Saccharomyces cerevisiae and purchased from Sigma-Aldrich Company in the United States; pNPG (p-nitrophenyl-β-D-galactopyranoside) was purchased from the United States Sigma-Aldrich Company; other reagents are analytically pure; the biologically active polypeptide synthesized in Example 1.

[0062] Experimental principle: α-glucosidase catalyzes pNPG to produce p-nitrophenol, which has a characteristic absorption peak at 405nm, so we use the kinetic / time software of the UV spectrophotometer to measure the UV absorption peak at 405nm, which can be displayed digitally The biologically active polypeptide synthesized in Example 1 inhibits the effect of α-glucosidase.

[0063] At 37°C, in a phosphate (0.05mol / L) buffer system of pH 6.8, 200μL α-glucosidase (0.175U / ml) and 100μL sericin peptide at different concentrations were incub...

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Abstract

The invention discloses sericin peptide with alpha-glycosidase inhibiting activity. The sequence of the sericin peptide is as shown in SEQ ID No: 1. The invention further relates to a synthesis preparation method of the sericin peptide with the alpha-glycosidase inhibiting activity and the application of biological activity of the sericin peptide. The sericin peptide disclosed by the invention has an important significance on food development, a health care product and a medicine which are capable of reducing postprandial blood sugar and development of food, a health care product and a medicine which are used for treating type 2 diabetic mellitus.

Description

technical field [0001] The invention relates to the field of polypeptide preparation in biotechnology, in particular to a sericin peptide with alpha-glucosidase inhibitory activity and its preparation and application. Background technique [0002] Bioactive peptides, as bioactive ingredients that promote health, have a variety of human metabolism and physiological regulation functions, are easy to digest and absorb, and have the functions of promoting immunity, hormone regulation, antibacterial, antiviral, lowering blood pressure, and lowering blood fat. A hot research topic and a functional factor with great development prospects. Various active peptides are often used as functional food additives in the practical production of food or medicine, especially in the application of milk drinks, food additives, and health foods, and have achieved good results. [0003] In recent years, with the improvement of people's living standards, the incidence of diabetes is showing an in...

Claims

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Application Information

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IPC IPC(8): C07K14/435C07K1/16A61K38/17A61P3/10A23L33/18
CPCA23L33/18A23V2002/00A61K38/00C07K14/43586A23V2200/30A23V2200/328Y02P20/55
Inventor 吴金鸿杨迎马鸿承陈姗姗张莉罗静王正武龚盛祥张维
Owner SHANGHAI JIAO TONG UNIV
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