Preparation method and application of microbial-source antioxidant protease

A microbial source, protease technology, applied in the field of genetic engineering and biological enzymes, can solve the problem of low yield and achieve good technical support, high application value, and wide application scenarios

Inactive Publication Date: 2017-10-17
ZHEJIANG FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Most of the existing antioxidant prote

Method used

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  • Preparation method and application of microbial-source antioxidant protease

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Embodiment Construction

[0024] The technical scheme of the present invention will be further explained by the following examples in conjunction with the accompanying drawings, but the protection scope of the present invention is not limited in any form by the examples.

[0025] 1. Construction of recombinant expression bacteria

[0026] (1) Use specific primers (SEQ ID NO.2 and SEQ ID NO.3) to amplify the genomic DNA of the Listeria wild strain (such as: EGDe reference strain or ATCC10403S standard strain), and simultaneously amplify The restriction sites of NdeI and XhoI are designed on the primers, and the amplification primers are:

[0027] Upstream primer: 5'-TTTCATATGGTAAAAAGAAATTACAGATGCAACATTTG-3'

[0028] Downstream primer: 5'-CCGCTCGAGAACGTATTTGTTGATGACTTCATCCAGTTC-3'

[0029] The PCR amplification system is: KOD-plus-Neo 1μL, 10×PCR Buffer for KOD-plus-Neo 5μL, MgSO 4 3 μL, dNTPs 5 μL, upstream primer 1 μL, downstream primer 1 μL, DNA template 2 μL, add ddH 2 0 to 50 μL. The PCR ampli...

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Abstract

The invention relates to the technical field of gene engineering and biological enzyme with an aim to provides a preparation method and application of microbial-source antioxidant protease. The amino acid sequence of the protease is as indicated in SEQ ID NO.1. The invention further the preparation method of the protease and the application of the protease in preparing drugs or health food used for inhibiting body aging and cell apoptosis. The protease is the important protease for oxidoreduction balance adjustment and plays an important role in biological oxidative stress and damage repair process, thereby being potential in application value in the anti-aging field. Through combination and regulation of factors of ASK1, NF-k and the like, body ageing and cell apoptosis are inhibited, cells can be subjected to damage repair and proliferation, cell ageing and cell apoptosis can be postponed, and the protease is one of important targets in tumor and cancer treatment; the protease is soluble in physical characteristic, natural activity is retained, and wide application prospect is achieved.

Description

technical field [0001] The invention belongs to the technical fields of genetic engineering and biological enzymes, and in particular relates to the preparation and application of a microbial source antioxidative protease. Background technique [0002] Microbial antioxidant protease is a kind of protein that widely exists in microorganisms in nature. The protein has enzymatic activity and has various functions such as maintaining redox balance in organisms, regulating biological signal transduction, DNA and protein damage repair, etc. Due to its rich source, easy purification, small molecular weight, and simple function, this protease was developed earlier in enzymatic research, and it also has a wide range of reference and application values ​​for drug preparation. At present, in the process of studying antioxidant enzymes, microbial-derived antioxidant proteases have powerful antioxidant effects, and can maintain intracellular redox homeostasis, anti-aging, and intestinal...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12N15/70A61K38/44A61P39/06A61K8/66A61Q19/08
CPCA61K8/66A61K38/00A61Q19/08C12N9/0051C12N15/70C12Y108/01008
Inventor 宋厚辉程昌勇叶美伶赵碧波邱旸汪渤森王晓杜
Owner ZHEJIANG FORESTRY UNIVERSITY
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