A group of DNA molecules and recombinant plasmids thereof as well as recombinant escherichia coli and application of recombinant escherichia coli in producing astragalin

A technology of DNA molecules and recombinant plasmids, applied in the field of genetic engineering technology and biomedicine, can solve the problems of low content, high extraction cost, environmental pollution, etc., and achieve the effect of simple process

Active Publication Date: 2016-11-09
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the relatively low content of ascarin in plants and the complex composition of

Method used

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  • A group of DNA molecules and recombinant plasmids thereof as well as recombinant escherichia coli and application of recombinant escherichia coli in producing astragalin
  • A group of DNA molecules and recombinant plasmids thereof as well as recombinant escherichia coli and application of recombinant escherichia coli in producing astragalin
  • A group of DNA molecules and recombinant plasmids thereof as well as recombinant escherichia coli and application of recombinant escherichia coli in producing astragalin

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Experimental program
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Embodiment 1

[0033] 1. Screening of Glycosyltransferase Genes

[0034] 1.1 Synthesis of glycosyltransferase gene

[0035] Glycosyltransferase 78D2 (NP_197207.1), 73B3 (NP_567953.1) and 78D1 (NP_564357.1) genes derived from Arabidopsis thaliana, glycosyltransferase UFGT (AEQ33588) derived from Ginkgo biloba .2) Gene and the glycosyltransferase BTGT (AGH18136.1) gene derived from Bacillus sp.HH1500, according to the Escherichia coli K12 dominant codon table (http: / / www.kazusa.or.jp / codon / ) for the above-mentioned gene The sequence is optimized, and codons with high usage frequency of amino acid synonymous codons are selected as dominant codons, and finally the optimized glycosyltransferase gene sequence information is obtained, which is synthesized by Shanghai Jierui Bioengineering Co., Ltd. to obtain the optimized glycosyl The nucleic acid sequences of the transferase genes op78D2, op73B3, op78D1, opUFGT, opBTGT are shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 and SEQ ID NO:...

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Abstract

The invention discloses a group of DNA molecules and recombinant plasmids thereof as well as recombinant escherichia coli and an application of the recombinant escherichia coli in producing astragalin. The DNA molecules include an arabidopsis glycosyltransferase gene 78D2, a sucrose phospholylase gene Basp, a uridyltransferase ugpA and a sucrose permease gene cscB which are optimized; and the nucleotide sequences of the genes are separately shown as SEQ ID NO:1, SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:6. The recombinant bacterium (the recombinant escherichia coli) can achieve the efficient expression of complete-gene optimized glycosyltransferase gene op78D2, and glycosyltransferase can efficiently and specifically conduct glycosylation on kaempferol 3rd-site hydroxyl; and meanwhile, by enhancing a UDP-glucose synthesis route of the recombinant bacterium, the recombinant bacterium can efficiently supply UDP-glucose, so that the bottleneck of the recombinant bacterium in synthesizing the astragalin is relieved. Finally, by optimizing fermentation conditions, the efficient synthesis of the astragalin is achieved.

Description

technical field [0001] The invention belongs to the field of genetic engineering technology and biomedicine, and specifically relates to a group of DNA molecules and their recombinant plasmids, recombinant Escherichia coli and their application for producing astragalus glycosides. Background technique [0002] Ascarin is a flavonoid found in a variety of plants. Such as lotus leaf, mulberry leaf, Tianshan Sorbus leaf, Uyghur sunflower and so on. The chemical name of ascarin is 3,5,7,4'-tetrahydroxyflavone-3-glucoside, and the molecular formula is C 21 h 20 o 11 , the relative molecular weight is 448.38, and its molecular structure is shown in Figure-1. It has various pharmacological effects such as significant antioxidant activity, anti-inflammatory activity, anti-allergy, and regulation of body immunity. [0003] Anti-inflammatory effect: Astragalus can significantly inhibit the production of NO, PGE2 and IL-6 or TNF-α induced by LPS in RAW264.7 cells; studies have sho...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N15/70C12N1/21C12P19/60C12R1/19
CPCC12N9/1048C12N9/1051C12N9/1241C12N15/70C12N2800/101C12N2800/22C12P19/60C12Y204/01007
Inventor 裴建军董萍赵林果解静聪
Owner NANJING FORESTRY UNIV
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