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Preparation method and application of β-picric acid

A picric acid and nucleotide technology, applied in the direction of using vectors to introduce foreign genetic material, recombinant DNA technology, fungi, etc., can solve uneconomical problems

Active Publication Date: 2017-06-23
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the content of picric acid in beer is very low, and adults need to take at least about 150 mg per day in order to achieve health care. Increasing the content of picric acid in beer is a difficult practical production problem

Method used

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  • Preparation method and application of β-picric acid
  • Preparation method and application of β-picric acid
  • Preparation method and application of β-picric acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1. The influence of PT1 and PT2 genes on the synthesis of β-picric acid after optimization of the preferred codons in the genomes of different species

[0066] 1. Preparation of recombinant yeast cells

[0067] A method for constructing recombinant yeast cells, comprising the step of transforming recipient cells into S1), S2), S3) and S4) to obtain recombinant cells:

[0068] S1) introducing picric acid synthesis-related gene 1 into the recipient cell;

[0069] S2) introducing picric acid synthesis-related gene 2 into the recipient cell;

[0070] S3) introducing a short side chain acyl-CoA ligase gene into the recipient cell;

[0071] S4) Introducing the valerophenone synthase gene into the recipient cells.

[0072] The short side chain acyl-CoA ligase genes are the short side chain acyl-CoA ligase CCL2 gene and the short side chain acyl-CoA ligase CCL4 gene. The nucleotide sequence of picric acid synthesis-related gene 1 is SEQ ID No.1 in the sequence listi...

Embodiment 2

[0098] Example 2, Effects of different fragments of picric acid synthesis-related gene 1 and different fragments of picric acid synthesis-related gene 2 on the synthesis of β-picric acid

[0099] 1. Effects of different fragments of picric acid synthesis-related gene 1 on the synthesis of β-picric acid

[0100] The experiment was repeated three times, and the specific steps of each repeated experiment were as follows:

[0101] Deleting 40, 60, 80, 100, 120, and 140 amino acids at the N-terminal of picric acid synthesis-related protein 1, respectively, to obtain picric acid synthesis-related protein 1Δ40 (amino acid sequence such as the 41st- amino acid 414), picric acid synthesis-related protein 1Δ60 (amino acid sequence shown in amino acid 61-414 of SEQ ID No.11), picric acid synthesis-related protein 1Δ80 (amino acid sequence shown in SEQ ID No.11 amino acid 81-414 amino acids), picric acid synthesis-related protein 1Δ100 (amino acid sequence is shown in amino acid 101-414 ...

Embodiment 3

[0122] The synthesis of embodiment 3, β-picric acid

[0123] The experiment was repeated three times, and the specific steps of each repeated experiment were as follows:

[0124] The pESC-LEU / CCL2 / CCL4 and pESC-HIS / VPS in step 1 of Example 1 were respectively introduced into yeast cell DD104 to obtain the coding sequence containing the short side chain acyl-CoA ligase CCL2, the short side chain acyl-CoA ligase The recombinant yeast cell of the coding sequence of ligase CCL4 and the coding sequence of valerophenone synthase was named DD104-CCL2 / CCL4 / VPS.

[0125] The sequence between the BamH I and Sal I recognition sites of the pESC-URA vector is replaced with the DNA molecule (picric acid synthesis-related gene 1Δ100) shown in the 301-1245 nucleotides of SEQ ID No.1, keeping other sequences unchanged. Transform to obtain the recombinant vector pESC-URA / PT1Δ100-At, pESC-URA / PT1Δ100-At, pESC-LEU / CCL2 / CCL4 and pESC-HIS / VPS in Step 1 of Example 1 were introduced into the yeast c...

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Abstract

The invention discloses a preparation method and application of beta-picric acid. The preparation method of beta-picric acid comprises a construction method of reconstituted cells, wherein the construction method of the reconstituted cells comprises steps (1), (2), (3) and (4) to reconstruct receptor cells so as to obtain the reconstituted cells: (1) introducing a picric acid synthesis related gene 1 into the receptor cells; (2) introducing a picric acid synthesis related gene 2 into the receptor cells; (3) introducing a short side chain ester-acyl coenzyme A ligase gene into the receptor cells; and (4) introducing a valerophenone synthetase gene into the receptor cells, wherein the picric acid synthesis related gene 1 is used for coding picric acid synthesis related protein 1, the picric acid synthesis related gene 2 is used for coding picric acid synthesis related protein 2, and the receptor cells are microbial cells, non-human animal cells or plant cells.

Description

technical field [0001] The invention relates to a preparation method and application of β-picric acid in the field of biotechnology. Background technique [0002] Hops (Humulus lupulus L.), also known as Humulus, hops, etc., are diploid (2n=20), belong to Rosaceae Cannabis Humulus hop species, are perennial climbing herbs, dioecious, The female flowers are arranged in a cone-shaped inflorescence by bracts, 3-4 cm long, and a large number of yellow glandular hairs grow under the inner bracts. The female flowers of hops are mainly used in brewing beer, and are the main source of the unique aroma and bitterness of beer, and are generally considered to be the "green gold" of beer brewing. In 2010, the global beer sales were 585 billion US dollars, and the total sales in 2015 are expected to reach 630 billion US dollars (https: / / uk.finance.yahoo.com / news / global-beer-industry-000000008.html), whether domestic Abroad, the beer sales market is still expanding. Whether the beer is...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/81C12N1/19C12P7/26
Inventor 王国栋李好勋覃浩班兆男
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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