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[Alpha]-bisabolol synthetic plasmid as well as construction method and escherichia coli engineered strain thereof

A technology of bisabolol and Escherichia coli, applied in the field of metabolic engineering, can solve the problems of complex chiral chemical structure, disappearance of endangered plants, production scale endangering ecological and environmental safety, etc.

Active Publication Date: 2018-07-20
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as a secondary metabolite, α-bisabolol has a limited natural yield. Obtaining α-bisabolol by direct extraction from biomass is still subject to many factors such as organism growth cycle, environment, and region, and the production The expansion of the scale will endanger the ecological and environmental safety, and even cause the ecological disaster of the disappearance of endangered plants
If chemical synthesis is adopted, the complex chiral chemical structure of α-bisabolol makes direct chemical synthesis difficult, with low purity and poor biological activity

Method used

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  • [Alpha]-bisabolol synthetic plasmid as well as construction method and escherichia coli engineered strain thereof
  • [Alpha]-bisabolol synthetic plasmid as well as construction method and escherichia coli engineered strain thereof
  • [Alpha]-bisabolol synthetic plasmid as well as construction method and escherichia coli engineered strain thereof

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Embodiment 1

[0046] Example 1: α-bisabolol synthetic plasmid and its construction method and Escherichia coli engineering strain

[0047] 1. α-bisabolol synthetic plasmid and its construction

[0048] The synthetic plasmid of α-bisabolol is based on the pTrc99A vector as the backbone plasmid, and the E. coli gene is assembled on the backbone plasmid ispA And a synthetic α-bisabolol synthase gene or wild-type α-bisabolol synthase gene from a different source; the synthetic α-bisabolol synthase gene is selected from codon-optimized Synthetic type AaBBS Gene, synthetic type LdBBS Gene, synthetic type MrBBS Gene, synthetic type ScBBS Gene and synthetic type SsBBS For any one of the genes, the nucleotide sequence is as shown in the sequence table SEQ ID No. 1, SEQ ID No. 2, SEQ ID No. 3, SEQ ID No. 4, and SEQ ID No. 5, correspondingly, The α-bisabolol synthetic plasmids were named pT-IAa, pT-ILd, pT-IMr, pT-ISc and pT-ISs, respectively; the wild-type α-bisabolol synthase gene was selected fr...

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Abstract

The invention discloses an [alpha]-bisabolol synthetic plasmid as well as a construction method and an escherichia coli engineered strain thereof. An IPP and DMAPP synthetic plasmid pS-MVA and the [alpha]-bisabolol synthetic plasmid are constructed, and the two plasmids are introduced into an escherichia coli strain E.coli DH5[alpha]. The [alpha]-bisabolol synthetic plasmid takes a pTrc99A vectoras a skeleton plasmid and is assembled with an escherichia coli gene ispA and an artificially synthetic [alpha]-bisabolol synthetase gene or a wild-type [alpha]-bisabolol synthetase gene from different sources; the artificially synthetic [alpha]-bisabolol synthetase gene is selected from one of AaBBS, LdBBS, MrBBS, ScBBS and SsBBS; and the wild-type [alpha]-bisabolol synthetase gene is selected from one of LdBBS, ScBBS and MrBBS. According to the invention, the [alpha]-bisabolol is synthesized through a biological method; and the [alpha]-bisabolol is high in yield, high in activity and is green-going and environment-friendly.

Description

Technical field [0001] The invention belongs to the field of metabolic engineering, and specifically relates to an alpha-bisabolol synthetic plasmid, a construction method thereof, and an E. coli engineering strain and application. Background technique [0002] Alpha-bisabolol (bisabolol), also known as bisabolol, is a natural sesquiterpene alcohol that exists in nature. It has several optical isomers as follows: [0003] [0004] They play an important role in medicine, food and cosmetics industries. (-)-α-Bisabolol mainly exists in the chrysanthemum chrysanthemum ( Matricaria recutita ) And other plants, have antibacterial, anti-inflammatory, anti-microbial and anti-digestive effects. It is used as an active ingredient in skin cosmetics or to inhibit skin inflammation to protect and care for allergic skin. [0005] Native American herb sweet diffusa ( Lippia dulcis Hernandulcin contained in) is a natural high-efficiency sweetener whose main chain is (+)-α-bisabolol. Henan Duxi...

Claims

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Application Information

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IPC IPC(8): C12N15/66C12N15/70C12N1/21C12P7/22C12R1/19
CPCC12N15/66C12N15/70C12P7/22
Inventor 王崇龙金善元
Owner SUZHOU UNIV
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