Lactic acid bacterium containing isoflavone synthetase (IFS)-cytochromes P450 reductase (CPR) fusion gene and application thereof

A technology of isoflavones and Lactococcus lactis, applied in the direction of bacteria, microorganism-based methods, enzymes, etc., can solve the problems of high cost and low yield, and achieve the effect of a huge potential market

Inactive Publication Date: 2012-03-21
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Separation of soybean isoflavones from soybean meal is the main way to obtain them in industry, and the cost is high and the yield is low.

Method used

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  • Lactic acid bacterium containing isoflavone synthetase (IFS)-cytochromes P450 reductase (CPR) fusion gene and application thereof
  • Lactic acid bacterium containing isoflavone synthetase (IFS)-cytochromes P450 reductase (CPR) fusion gene and application thereof
  • Lactic acid bacterium containing isoflavone synthetase (IFS)-cytochromes P450 reductase (CPR) fusion gene and application thereof

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Embodiment 1

[0041] Embodiment 1, construction and application of genetically engineered lactic acid bacteria containing isoflavone synthase-cytochrome P450 reductase fusion gene

[0042] 1. Construction of fusion gene IFS-CPR

[0043] 1. Acquisition of soybean isoflavone synthase gene (IFS)

[0044] (1) UV-induced North my country soybean (Glycine max L.Merr.) cultivar spring soybean series (Beifeng 12) (the public can obtain from China Agricultural University, the non-patent literature that has recorded this material is: Hao Jia, Ma Huiqin, Chen Shangwu. Using the SMART method to rapidly clone multiple genes of the isoflavone metabolic pathway. Chinese Journal of Biotechnology, 2007, 27(6): 66-70) Expression of soybean isoflavone synthase gene (IFS) in young shoots, extraction of soybean Sprout total RNA.

[0045] (2) reverse transcription into cDNA library.

[0046] (3) Design specific primers and obtain IFS gene by PCR amplification

[0047] The upstream primer is IFS-F: 5'-GtctagaAT...

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Abstract

The invention discloses lactic acid bacterium containing an IFS-CPR fusion gene and application thereof. The recombinant bacterium provided in the invention is recombinant bacterium obtained by introducing coding genes of IFS-CPR into host bacterium, and IFS-CPR is a protein as described in 1) or 2): 1) a protein as represented by sequence 1 in a sequence table; 2) a protein derived by substituting and/or deleting and/or adding one or more amino acid residue in the amino acid residue sequence of sequence 1 in the sequence table. A recombinant lactic acid bacterial strain obtained in the invention can be used for production of isoflavone and has a great potential market, economic benefits and social benefits in the food industry.

Description

technical field [0001] The invention relates to a recombinant bacterium and its application, in particular to a lactic acid bacterium containing an isoflavone synthase-cytochrome P450 reductase fusion gene and its application. Background technique [0002] Isoflavones are an important class of phytoestrogen-like active substances mainly present in soybeans and other leguminous plants, including daidzin, genistin and glycitin, etc. Its functional components are aglycone moieties, mainly daidzein and genistein. The chemical name of genistein is 4,5,7-trihydroxyisoflavone, and the molecular formula is C 15 h 10 o 5 , the molecular weight is 270.24, the CAS number is 446-72-0, and the structural formula is formula 1; the chemical name of daidzein is 7-hydroxy-3-(4-hydroxyphenyl)-4-benzopyrone, and the molecular formula is: C 15 h 10 o 4 , molecular weight: 254.24, CAS number: 486-66-8, structural formula is formula 2. [0003] [0004] Formula 1 Formula 2 [0005] Me...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/62C12N9/00C12P17/06C12R1/46
Inventor 陈尚武左芳雷卢晓明赵建云郑超郝彦玲李平兰马会勤
Owner CHINA AGRI UNIV
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