Shuimoliuyun (Talk | contribs) |
Shuimoliuyun (Talk | contribs) |
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to acrylic acid is a "roof" factor that restricts high yield of acrylic acid. | to acrylic acid is a "roof" factor that restricts high yield of acrylic acid. | ||
<br> We chose E. coli and S. cerevisiae, the two most convenient model chassis organisms in prokaryotic | <br> We chose E. coli and S. cerevisiae, the two most convenient model chassis organisms in prokaryotic | ||
− | and eukaryotic organisms. In order to | + | and eukaryotic organisms. In order to investigate the chassis cells’ tolerance to acrylic acid, we |
set up a cytotoxicity test where the two chassis cells grew in different concentrations of acrylic | set up a cytotoxicity test where the two chassis cells grew in different concentrations of acrylic | ||
acid medium, and the bacteria OD changes were monitored.The results are as follows: | acid medium, and the bacteria OD changes were monitored.The results are as follows: | ||
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<br> Fig1. OD of E.coli MG1655 under acrylic acid of different concentration and time | <br> Fig1. OD of E.coli MG1655 under acrylic acid of different concentration and time | ||
− | <br> Fig2. OD of S. | + | <br> Fig2. OD of S. cerevisiae BY4741 under acrylic acid of different concentration and time |
<br> Two kinds of chassis cells have different tolerance to acrylic acid. Here we selected 500mg / L | <br> Two kinds of chassis cells have different tolerance to acrylic acid. Here we selected 500mg / L | ||
and 1000mg / L two kinds of acrylic acid concentration to analyze: | and 1000mg / L two kinds of acrylic acid concentration to analyze: | ||
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<br> | <br> | ||
<br> It can be seen that, similar to the results of E. coli, the introduction of new pathways does improve | <br> It can be seen that, similar to the results of E. coli, the introduction of new pathways does improve | ||
− | the ability of S. cerevisiae synthesizing acrylic acid. Compared the old pathway introduced only | + | the ability of S. cerevisiae synthesizing acrylic acid. Compared with the old pathway introduced only |
ceaS2 enzyme, acrylic acid production was increased by 3 times after introduction of GlyDH enzymes | ceaS2 enzyme, acrylic acid production was increased by 3 times after introduction of GlyDH enzymes | ||
and DAK enzymes. And the yield of acrylic acid was increased by 5 times compared to the old pathway | and DAK enzymes. And the yield of acrylic acid was increased by 5 times compared to the old pathway | ||
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genes: | genes: | ||
<br> | <br> | ||
− | <br> Fig XX S.C | + | <br> Fig XX S.C BY4741's DLD1 gene Agarose gel figure of colonies verification after CRISPR knockout. |
<br> Wt is the corresponding nucleic acid stripe of wild-type S.C BY4741; M is a GeneRuler 1 kb DNA ladder; | <br> Wt is the corresponding nucleic acid stripe of wild-type S.C BY4741; M is a GeneRuler 1 kb DNA ladder; | ||
lanes 1, 2, 3 are three selected nucleic acid stripes of monoclonal colonies. | lanes 1, 2, 3 are three selected nucleic acid stripes of monoclonal colonies. | ||
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<br> | <br> | ||
<br> It can be seen that the optimization of bypass metabolic flux is conducive to the concentration | <br> It can be seen that the optimization of bypass metabolic flux is conducive to the concentration | ||
− | of metabolic flux and improving the yield of acrylic acid. Of | + | of metabolic flux and improving the yield of acrylic acid. Of course we also found in the process |
of the experiment that after knocking out the 9 genes, S. cerevisiae colony growth became very slow, | of the experiment that after knocking out the 9 genes, S. cerevisiae colony growth became very slow, | ||
indicating that a more tender method should be adopted, such as RNAi, to inhibit the bypass pathway. | indicating that a more tender method should be adopted, such as RNAi, to inhibit the bypass pathway. |
Revision as of 11:39, 1 November 2017