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Hypoglycemic polypeptide and preparation method and application thereof

A technology of hypoglycemic polypeptide and glucosidase, which is applied to the hypoglycemic polypeptide that inhibits the activity of α-glucosidase. The preparation of the hypoglycemic polypeptide and the field of hypoglycemic polypeptide can solve the problems that have not been found in the research of carrot polypeptide, and achieve easy Operation and implementation, simple preparation, and the effect of increasing the added value of production

Active Publication Date: 2019-10-11
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing research on carrots mainly focuses on β-carotene, pectin, quercetin, phenols, etc., and research on carrot polypeptides has not yet found

Method used

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  • Hypoglycemic polypeptide and preparation method and application thereof
  • Hypoglycemic polypeptide and preparation method and application thereof
  • Hypoglycemic polypeptide and preparation method and application thereof

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preparation example Construction

[0058] The sixth aspect of this specification provides a method for preparing the gene as described in the second aspect, comprising the following steps:

[0059] With the nucleotide sequence shown in SEQ ID No: 7-12 as a template, the sequence shown in SEQ ID No: 13 is added at its 5' end as a forward primer, and the sequence shown in SEQ ID No: 14 is added at its 3' end. The sequence shown is the reverse primer, and PCR amplification is carried out to obtain the gene.

[0060] The seventh aspect of this specification provides a method for preparing the hypoglycemic polypeptide as described in the first aspect, which is extracted from natural carrots, including the following steps:

[0061] Dice the carrots, squeeze the juice, centrifuge, filter, and collect the concentrated carrot juice for sample;

[0062] Collect the eluted samples after loading the concentrated carrot juice through an anion exchange column for separation;

[0063] After concentrating the eluted sample, ...

Embodiment 1

[0068] Example 1: Enrichment and preparation of natural carrot hypoglycemic polypeptide

[0069] A method for preparing a hypoglycemic polypeptide is extracted from natural carrots, comprising the following steps:

[0070] Dice the carrots, add 1 volume of distilled water, and squeeze the juice with a juicer at room temperature. After collecting the juice for the first time, centrifuge at 13000rpm for 30min on a high-speed centrifuge to collect the supernatant. The collected supernatant was left standing at 4° C. for 12 h to allow the residual pomace in the supernatant to settle. The settled supernatant was centrifuged again at 13000 rpm for 30 min, and the centrifuged supernatant was collected. Add pectinase powder to the supernatant after the second centrifugation according to the weight ratio of pectinase powder to 1:10000, stir evenly, and place at 37° C. for 2 hours. After filtering the supernatant added with pectinase powder through filter paper, the filtrate was cent...

Embodiment 2

[0073] Example 2: Preparation of recombinant hypoglycemic polypeptide DCHP

[0074] The preparation of the recombinant hypoglycemic polypeptide DCHP is based on the amino acid sequence table SEQ ID NO: 1 of the natural hypoglycemic polypeptide DCHP, through base optimization, chemically synthesizes a coding gene suitable for expression in Escherichia coli, and expresses and purifies it in Escherichia coli to obtain Recombinant hypoglycemic polypeptide DCHP with α-glucosidase inhibitory activity. Specifically include the following steps:

[0075] According to the amino acid sequence SEQ ID NO: 1 of the natural hypoglycemic polypeptide obtained in Example 1, chemically synthesize the nucleotide sequence SEQ ID NO: 7 encoding the natural hypoglycemic polypeptide, and then add The forward primer of BamHI is shown in SEQ ID No: 13, the reverse primer of EcoRI added at the 3' end is shown in SEQ ID No: 14, and then the synthetic gene is used as a template for PCR amplification. Th...

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Abstract

The invention discloses a hypoglycemic polypeptide and a preparation method and application thereof. The hypoglycemic polypeptide has the activity of inhibiting alpha-glucosidase and has a following amino acid sequence of (1) or (2), wherein an amino acid sequence shown in SEQ ID No:1 is involved in the (1), the amino acid sequence in the (2) has at least 80% homology with that in (1) and has thefunctions of the amino acid sequence in the (1). The hypoglycemic polypeptide can be obtained through natural isolation enrichment and artificial polypeptide synthesis or through prokaryocyte or eukaryocyte expression purification. In addition, the hypoglycemic polypeptide has high tolerance to trypsin and chymotrypsin, has great significance for the prevention and treatment of diabetes, and alsoprovides a new direction for increasing the production added value of carrots.

Description

technical field [0001] The present invention belongs to the field of biopharmaceuticals, and specifically relates to a hypoglycemic polypeptide, in particular to an orally-available hypoglycemic polypeptide extracted from carrots that inhibits the activity of α-glucosidase, and the present invention also relates to the preparation of the hypoglycemic polypeptide methods and applications. Background technique [0002] As a complex chronic metabolic disease, diabetes has many complications, which will have a significant impact on the nervous system, urinary system and visual system of the human body. For the clinical treatment of diabetes, it mainly includes two aspects, one is the injection of insulin that can lower blood sugar; the other is oral hypoglycemic drugs, mainly including: insulin secretagogues, α-glucosidase inhibitors, biguanides and thiazolidinediones are four oral hypoglycemic agents. α-glucosidase inhibitor delays or prevents the hydrolysis of polysaccharide...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/11C12N15/10C07K1/18A23L33/18A61K38/16A61P3/10
CPCC07K14/415A23L33/18A61P3/10A23V2002/00A61K38/00A23V2200/328A23V2250/55
Inventor 石亚伟郝杨吴峰王蕾
Owner SHANXI UNIV
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