Short-chain dehydrogenase BLSDR1 as well as coding gene and application thereof

A short-chain dehydrogenase, gene technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problems of volatility and toxicity, respiratory system toxicity and other issues

Active Publication Date: 2021-07-23
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition to formaldehyde, there are many volatile organic compounds that can cause pollution. For example, hexanal, acetaldehyde, acetone, etc., which are usually at relatively high concentrations in the indoor environment, due to their volatility and toxicity, will have a negative impact on the human respiratory system. Potentially toxic effects

Method used

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  • Short-chain dehydrogenase BLSDR1 as well as coding gene and application thereof
  • Short-chain dehydrogenase BLSDR1 as well as coding gene and application thereof
  • Short-chain dehydrogenase BLSDR1 as well as coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Acquisition of Bacillus coagulans NL01 short-chain dehydrogenase gene, construction of recombinant plasmid and transformation of Escherichia coli.

[0023] The genomic DNA of Bacillus coagulans NL01 was prepared by a conventional method. For this process, the genomic DNA of Bacillus coagulans NL01 can be extracted by referring to the instructions of the Takara MiniBEST Universal Genomic DNA Extraction Kit Ver.5.0.

[0024] Design primers and introduce restriction sites that can be inserted into BamHI and HindIII of plasmid pETDuet-1. The designed primer sequences of blsdr1 are as follows:

[0025] Upstream primer 5'-GGCCTTGGATCCGatgcaaacatttgatttcacaggaaaagttgcaatcgtaa-3' (Bam HI)

[0026] Downstream primer 5'-AATTTTTAAAAGCTTttaggaaaggactgccccgccatcaatgacaaccg-3'(HindIII)

[0027] Using the extracted genome of Bacillus coagulans NL01 as a template, the above primers were used for amplification, and the short-chain dehydrogenase gene sequence blsdr1 was obtai...

Embodiment 2

[0030] Example 2: Induced expression of recombinant short-chain dehydrogenase.

Embodiment 1

[0031] The engineered bacteria E.coli BL21(DE3) / pETDuet-blsdr1 constructed in Example 1 was inoculated into LB liquid medium containing 50ug / mL ampicillin, cultivated overnight at 37°C, and then inoculated with 1% inoculum size (v / v) Into 50mL LB medium containing 50ug / mL ampicillin, culture at 37°C, 200rpm to cell concentration OD 600 To about 0.6-0.8, add IPTG with a final concentration of 1mM, induce culture at 37°C for 5h, then centrifuge at 8000rpm at 4°C for 5min to collect the cells, and store them at -20°C for later use.

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Abstract

The invention discloses short-chain dehydrogenase BLSDR1 as well as a coding gene and application thereof. The protein provided by the invention is a protein coded by an amino acid sequence as shown in SEQ NO.2. A new short-chain dehydrogenase gene blsdr1 is cloned from bacillus coagulans NL01, the gene can be expressed in a host cell to produce the short-chain dehydrogenase, and the short-chain dehydrogenase has a wide substrate spectrum in the aspect of reducing carbonyl compounds, has certain organic solvent tolerance and can be used for removal and reduction of various ketoaldehyde compounds.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a short-chain dehydrogenase, an encoded gene and an application function. Background technique [0002] With the rapid development of society and urbanization, indoor air pollution has become a concern, among various pollutants, volatile organic compounds including aromatic compounds and formaldehyde are complex, irritating and pathogenic , It has a strong stimulating effect on human eyes, nose, skin, lungs, respiratory tract, etc., and some may cause cancer, so it has received widespread attention. In addition to formaldehyde, there are many volatile organic compounds that can cause pollution, for example, hexanal, acetaldehyde, acetone, etc., which are usually relatively high concentrations in indoor environments, due to their volatility and toxicity, can be harmful to the human respiratory system. cause potential toxic effects. [0003] Carbonyl compounds are one of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/04C12N15/53C12N15/70C12N1/21C12P7/02C12R1/19
CPCC12N9/0006C12N15/70C12P7/02Y02E50/10
Inventor 欧阳嘉卜重阳陶渊明付佳铭郑兆娟
Owner NANJING FORESTRY UNIV
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