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Recombinant escherichia coli and method for biosynthesizing diosmetin by utilizing recombinant escherichia coli

A recombinant Escherichia coli and biosynthetic technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, botany equipment and methods, etc., can solve difficult, unenvironmental friendly, low diosmin and diosmin content and other issues, to achieve the effect of improving soluble expression and good catalytic activity

Active Publication Date: 2021-04-30
HUNAN AGRI PRODS PROCESSING INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The main ways to obtain diosmin at this stage include: ① Obtained directly from plants through extraction and separation, but due to the content of diosmin and diosmin in plants Very low, direct extraction is costly and difficult, and has not been widely used
②In actual production at the present stage, diosmin and diosmin are obtained from hesperidin, which is relatively high in citrus fruits, by dehydrogenation under high heat conditions with pyridine as solvent and iodine and bromine as catalysts, but the The method requires the use of toxic reagents and a large amount of organic solvents, which is neither environmentally friendly nor safe

Method used

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  • Recombinant escherichia coli and method for biosynthesizing diosmetin by utilizing recombinant escherichia coli
  • Recombinant escherichia coli and method for biosynthesizing diosmetin by utilizing recombinant escherichia coli
  • Recombinant escherichia coli and method for biosynthesizing diosmetin by utilizing recombinant escherichia coli

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Embodiment 1

[0041] A recombinant Escherichia coli of the present invention, the recombinant Escherichia coli is obtained by introducing the recombinant plasmid pAnFNSⅠ into the host Escherichia coli, the recombinant plasmid is constructed by linking the codon-optimized AnFNSⅠ gene and the expression vector pET-28a(+), the code The amino acid sequence of AnFNSⅠ before codon optimization is shown in SEQ ID NO.1, and the amino acid sequence of AnFNSⅠ after codon optimization is shown in SEQ ID NO.4.

[0042] In this example, the nucleotide sequence encoding AnFNSⅠ before codon optimization is shown in SEQ ID NO.2, and the nucleotide sequence encoding AnFNSⅠ after codon optimization is shown in SEQ ID NO.5.

[0043] In this example, the host Escherichia coli is Escherichia coli BL21(DE3), and the recombinant Escherichia coli is designated as DE3 / pAnFNSI.

[0044] The construction method of the recombinant escherichia coli of the present embodiment is as follows:

[0045] (1) The coding gene ...

Embodiment 2

[0057] A recombinant Escherichia coli of the present invention, the recombinant Escherichia coli is obtained by introducing the recombinant plasmid pAnFNSⅠ into the host Escherichia coli, the recombinant plasmid is constructed by linking the codon-optimized AnFNSⅠ coding gene and the expression vector pET-28a(+), the code The amino acid sequence of flavone synthase AnFNSⅠ before codon optimization is shown in SEQ ID NO.1, and the amino acid sequence of flavone synthase AnFNSⅠ after codon optimization is shown in SEQ ID NO.4.

[0058] In this example, the nucleotide sequence encoding codon-optimized flavone synthase AnFNSⅠ is shown in SEQ ID NO.2, and the nucleotide sequence encoding codon-optimized flavone synthase AnFNSⅠ is shown in SEQ ID NO.5 Show.

[0059] In this embodiment, the host Escherichia coli is Escherichia coli BL21(DE3).

[0060] The construction method of the recombinant escherichia coli of the present embodiment is as follows:

[0061] The recombinant vector...

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Abstract

The invention discloses recombinant Escherichia coli and a method for biosynthesizing diosmetin by using the recombinant Escherichia coli, the recombinant Escherichia coli is obtained by introducing a recombinant plasmid into host Escherichia coli, and the recombinant plasmid is constructed by connecting an encoding gene of AnFNSI after codon optimization and an expression vector pET-28a (+); The AnFNSI is flavone synthase 1 derived from angelica archang lica, the amino acid sequence of the AnFNSI is as shown in SEQ ID NO.1, and the amino acid sequence of the AnFNSI after codon optimization is as shown in SEQ ID NO.4. The recombinant Escherichia coli is used as a whole-cell catalyst to catalyze a substrate hesperetin to biosynthesize diosmetin. The method is green and efficient, does not need to use a large amount of organic solvents or toxic chemical reagents in the reaction process, and is low in cost, high in safety and easy for large-scale production.

Description

technical field [0001] The invention belongs to the technical field of biochemical industry, and relates to a recombinant Escherichia coli and a method for biosynthesizing diosmin by using the recombinant Escherichia coli, in particular to a recombinant Escherichia coli and a highly efficient method for converting hesperetin into diosmin by using the recombinant Escherichia coli Biosynthetic methods. Background technique [0002] Diosmetin is a flavonoid compound isolated from plants, and its structural formula is as follows: [0003] [0004] Dianophyllin is mainly found in citrus, mint and other plants. The content of free diosmin in plants is low, about 0.027% in mint and about 0.016% in citrus. Diosmin mainly exists in the form of rutinoside-diosmin in plants. At present, diosmin and diosmin are widely used in clinical treatment as vascular protective drugs that enhance venous tone. Only diosmin tablets In 2016, sales of a drug retailed in domestic pharmacies reach...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/53C12N15/70C12P17/06C12R1/19
CPCC12N9/0071C12N15/70C12P17/06C12Y114/11022C12N2800/22
Inventor 单杨李高阳王振刘娟操文军
Owner HUNAN AGRI PRODS PROCESSING INST
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