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Method for preparing shikimic acid using biosynthesis technology and engineered bacteria

A technology for biosynthesis and shikimic acid, applied in the field of genetic engineering, can solve the problems of low shikimic acid synthesis rate and yield, low purity, etc., and achieves increased gluconeogenesis and glycogen synthesis, simple operation process, and reduced glycolysis. Effect

Inactive Publication Date: 2007-12-12
汪莉
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Problems solved by technology

[0009] In order to improve the defects of low shikimic acid synthesis rate, yield and low purity in the current method for synthesizing shikimic acid using genetic engineering technology, the present invention discloses a method for preparing shikimic acid using biosynthesis technology

Method used

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  • Method for preparing shikimic acid using biosynthesis technology and engineered bacteria
  • Method for preparing shikimic acid using biosynthesis technology and engineered bacteria
  • Method for preparing shikimic acid using biosynthesis technology and engineered bacteria

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Embodiment 1

[0042] The construction of embodiment one shikimic acid expression bacterial strain

[0043] Materials and Methods:

[0044] (1) Strains and plasmids. E.coli DH5α was purchased from Takara Company. Plasmid pKD46 (oriR101repA101tsP araB -gam-bet-exo Amp) is temperature sensitive and expresses the three γ-phage recombinases Gam, Bet and Exo required for homologous recombination after arabinose induction. Plasmid pKD3 is a chloramphenicol resistance gene containing FRT sites on both sides. Plasmid pCP20 is also a temperature-sensitive type. After heat induction, it expresses FLP recombinase, which can recognize FRT sites and promote the occurrence of recombination (1. Datsenko KA, Wanner BL. One step inactivation of chromosomal genes in Escherichia coli K 12 using PCR products. Proc Natl Acad Sci USA, 2000, 97: 6640~6645; 2. Research progress of Red recombination technology, Chinese Journal of Biotechnology, 2006, 26(1): 81~ 86), the above three plasmids were kindly donated b...

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Abstract

The invention discloses a method for preparing shikimic acid by using biological synthesis technique, and the employed recombined bacillus coli. The method comprises following steps: constructing carrier for speed- limit enzyme polygene box containing shikimic acid metabolic pathway; repalcing shiA gene with polygene box; removing shikimic acid kinase- mb in bacteria by using Red recombinant system; fermenting recombinant engineering bacteria and getting shikimic acid and purifying shikimic acid. The invention can increase expression amount for shikimic acid, reduces production cost and provides raw material for preparing medicine Duffy that can prevent influenza.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a method for preparing shikimic acid through biosynthesis technology, and also relates to engineering bacteria used in the method. Background technique [0002] In recent years, bird flu has been proved to be able to directly infect humans, with ferocious onset and high fatality rate. According to statistics, bird flu has infected more than 100 people, and more than half of them died. As of October 2005, more than ten countries and regions, including Cambodia and China, have experienced the H5N1 virus epidemic, and it is still spreading. At present, governments of various countries are actively responding to the possible large-scale outbreak of bird flu in humans. Influenza virus experts believe that a new influenza pandemic is inevitable and may not be far away. At a seminar hosted by the World Health Organization on July 4, 2005, scientists pointed out that bird flu is sti...

Claims

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Application Information

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IPC IPC(8): C12P7/42C12N15/09C12N1/21C12R1/19
Inventor 汪莉
Owner 汪莉
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