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− | < | + | <h2 style="text-align:center">Introduction</h2> |
<h4>The essence of biochemical synthesis is the catalytic reaction with enzyme as the catalyst. Creating | <h4>The essence of biochemical synthesis is the catalytic reaction with enzyme as the catalyst. Creating | ||
new biochemical reactions is an important research direction of synthetic biology. | new biochemical reactions is an important research direction of synthetic biology. | ||
− | <br> ceaS2, whose full name is N2-(2-carboxyethyl)arginine synthase2, is a kind of enzyme in Streptomyces | + | <br><br> ceaS2, whose full name is N2-(2-carboxyethyl)arginine synthase2, is a kind of enzyme in Streptomyces |
clavuligerus. The mentor of our team, Jiang Huifeng, has confirmed the new functions of ceaS2 with the help of TPP (Thiamine pyrophosphate) and magnesium ions, ceaS2 enzyme can catalyze the | clavuligerus. The mentor of our team, Jiang Huifeng, has confirmed the new functions of ceaS2 with the help of TPP (Thiamine pyrophosphate) and magnesium ions, ceaS2 enzyme can catalyze the | ||
production of acrylic acid with DHAP (dihydroxy acetone phosphate) or G3P (glyceraldehyde 3-phosphate) | production of acrylic acid with DHAP (dihydroxy acetone phosphate) or G3P (glyceraldehyde 3-phosphate) | ||
as substrate. | as substrate. | ||
− | <br> Cell factory of acrylic acid (GAACF) 1.0: | + | <br><br> Cell factory of acrylic acid (GAACF) 1.0: |
− | <br> DHAP and G3P are the central metabolic secondary products which can be easily found in various organisms. | + | <br><br> DHAP and G3P are the central metabolic secondary products which can be easily found in various organisms. |
They are the carbon flow nodes that must be passed in the glycerol metabolic pathway in most organisms. | They are the carbon flow nodes that must be passed in the glycerol metabolic pathway in most organisms. | ||
With ceaS2 enzyme as the core part, it is possible to create a new pathway to synthesize acrylic | With ceaS2 enzyme as the core part, it is possible to create a new pathway to synthesize acrylic | ||
acid from glycerol metabolic pathway in organisms and construct a cell factory with high yield of | acid from glycerol metabolic pathway in organisms and construct a cell factory with high yield of | ||
acrylic acid. | acrylic acid. | ||
− | <br> First, we took E. coli BL21 (DE3) as the chassis cells and constructed engineering bacteria carrying | + | <br><br> First, we took E. coli BL21 (DE3) as the chassis cells and constructed engineering bacteria carrying |
the gene of ceaS2 enzyme with pET-28a plasmid as the vector. We constructed a new pathway to synthesize | the gene of ceaS2 enzyme with pET-28a plasmid as the vector. We constructed a new pathway to synthesize | ||
acrylic acid from any carbon source by transforming ceaS2 directly into the chassis cells. This new | acrylic acid from any carbon source by transforming ceaS2 directly into the chassis cells. This new | ||
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as an example, we only need three enzymes to achieve the synthesis of acrylic acid from glycerol. | as an example, we only need three enzymes to achieve the synthesis of acrylic acid from glycerol. | ||
So this pathway is a more flexible and has more development prospects. | So this pathway is a more flexible and has more development prospects. | ||
− | <br> 【E.coli图+路径图+质粒图】 | + | <br><br> 【E.coli图+路径图+质粒图】 |
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− | Through the whole cell catalysis and HPLC (High Performance Liquid Chromatography), | + | <br>Through the whole cell catalysis and HPLC (High Performance Liquid Chromatography), |
the results show that the engineering bacteria can use glycerol as carbon source to produce acrylic | the results show that the engineering bacteria can use glycerol as carbon source to produce acrylic | ||
acid. However, the yield of the cell factory 1.0 is not high, only about 1mg / L. | acid. However, the yield of the cell factory 1.0 is not high, only about 1mg / L. | ||
− | <br> It is known that acrylic acid can not be metabolized in the cell, so we analyzed the possible reasons | + | <br><br> It is known that acrylic acid can not be metabolized in the cell, so we analyzed the possible reasons |
as the following: | as the following: | ||
<br> 1. The activity and the catalytic efficiency of wild type ceaS2 is low. | <br> 1. The activity and the catalytic efficiency of wild type ceaS2 is low. | ||
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</h4> | </h4> | ||
<div id="COREPART" style="padding-top:50px;margin-top:-50px;"> | <div id="COREPART" style="padding-top:50px;margin-top:-50px;"> | ||
− | <h2 style="text-align:center"> | + | <h2 style="text-align:center">Core Part</h2> |
<h4>Acrylic acid is a byproduct of CEAS2 enzyme, the catalytic effect of wild type ceaS2 enzyme is very | <h4>Acrylic acid is a byproduct of CEAS2 enzyme, the catalytic effect of wild type ceaS2 enzyme is very | ||
weak, and acrylic acid production is only 1mg / L. So it is necessary to improve the catalytic | weak, and acrylic acid production is only 1mg / L. So it is necessary to improve the catalytic |
Revision as of 13:07, 1 November 2017