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Polypeptides having deepoxidation catalytic activity, and encoding nucleic acids and application thereof

A catalytically active, epoxy-based technology, applied in the field of peptides, can solve problems such as dependence, harsh reaction conditions, and industrial application limitations.

Pending Publication Date: 2020-07-31
山东农之安生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, epoxy group-removed trichothecene derivatives can be chemically synthesized in vitro under high alkaline conditions, but it is difficult to apply in industry due to harsh reaction conditions and low efficiency
In addition, several anaerobic bacteria have been isolated from animal intestinal microorganisms that can detoxify trichothecenes, but the mechanism of action is not clear, and due to the dependence on anaerobic conditions, practical industrial applications have been greatly restricted. limit

Method used

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  • Polypeptides having deepoxidation catalytic activity, and encoding nucleic acids and application thereof
  • Polypeptides having deepoxidation catalytic activity, and encoding nucleic acids and application thereof
  • Polypeptides having deepoxidation catalytic activity, and encoding nucleic acids and application thereof

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Experimental program
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Embodiment

[0093] 1. Preparation of FTCD active polypeptide

[0094] 1. Materials and methods

[0095] Escherichia coli (Escherichia coli) DH5α strain, expression strain BL21(DE3), prokaryotic expression vector pET-28a(+) and plasmid pMD19-T-FTCD are preserved by our laboratory, among which the plasmid pMD19-T-FTCD contains The deepoxylase gene of wheatgrass, the sequence of which is shown in SEQ ID No.36.

[0096] 1.2 Experimental method

[0097] 1.2.1 The recombinant expression vector pET28a-FTCD was constructed by the following method.

[0098] According to expression vector pET28a design has the primer of NcoI and BamHI restriction site site, and primer sequence is as follows (underline indicates restriction restriction site):

[0099] Forward primer: 5'- CCATGG CTAGAAATCCACCCATCGTCATCACC-3'

[0100] Reverse primer: 5'- GGATCC TCTTCACCTCGGCATACTTGTC-3'

[0101] The plasmid pMD19-T-FTCD was used as a template for PCR amplification. The amplified product was detected by 1% ag...

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Abstract

The invention discloses a polypeptide having deepoxidation catalytic activity, and encoding nucleic acid and application thereof. The polypeptide provided by the invention can catalyze a reaction of trichothecene mycotoxins and glutathione under mild conditions to remove epoxy groups and generate non-toxic and harmless glutathione derivatives, so that detoxification and detoxification of trichothecene mycotoxins are realized. The polypeptide has wide applications in the fields of agriculture, food, feed, medicine and the like.

Description

technical field [0001] The present invention relates to the field of polypeptides, in particular to a polypeptide with catalytic activity for removing epoxy groups, its encoding nucleic acid and its application. Background technique [0002] The basic chemical structure of trichothecenes is a sesquiterpene, which is also called 12,13-epoxy trichothecenes because of the formation of epoxy groups on the 12th and 13th carbons compound. Since the 1970s, researchers have found that the epoxy group of trichothecenes is the main source of toxicity. At present, epoxy group-removed trichothecene derivatives can be chemically synthesized in vitro under high alkaline conditions, but it is difficult to be applied industrially due to harsh reaction conditions and low efficiency. In addition, several anaerobic bacteria have been isolated from animal intestinal microorganisms that can detoxify trichothecenes, but the mechanism of action is not clear, and due to the dependence on anaerobi...

Claims

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Application Information

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IPC IPC(8): C12N9/00C12N15/52C12P21/02C12Q1/25G01N33/68
CPCC07K5/0215C12N9/00C12P21/02C12Q1/25G01N33/68
Inventor 不公告发明人
Owner 山东农之安生物科技有限公司
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