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We also used the whole cell catalytic reaction and HPLC determination method to determine the amount of acrylic acid produced. | We also used the whole cell catalytic reaction and HPLC determination method to determine the amount of acrylic acid produced. | ||
For E. coli, yields of using new and old synthetic pathways of acrylic acid are as follows: | For E. coli, yields of using new and old synthetic pathways of acrylic acid are as follows: | ||
+ | <br> | ||
Conditions: reaction time 42h, PH8.0, glycerol concentration 1% | Conditions: reaction time 42h, PH8.0, glycerol concentration 1% | ||
<br> | <br> | ||
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Conditions: reaction time 72h, PH8.0, glycerol concentration 2% | Conditions: reaction time 72h, PH8.0, glycerol concentration 2% | ||
Normalized the results based on the acrylic acid yield of BY4741-ceas2 as the | Normalized the results based on the acrylic acid yield of BY4741-ceas2 as the | ||
− | indicator<br /> | + | indicator.<br /> |
<img src="https://static.igem.org/mediawiki/2017/2/2e/NPU-image17.png" style="max-width:60%;"><br /> | <img src="https://static.igem.org/mediawiki/2017/2/2e/NPU-image17.png" style="max-width:60%;"><br /> | ||
It can be seen that the optimization of bypass metabolic flux is conducive to the | It can be seen that the optimization of bypass metabolic flux is conducive to the | ||
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should be adopted, such as RNAi, to inhibit the bypass pathway.<br /> | should be adopted, such as RNAi, to inhibit the bypass pathway.<br /> | ||
</a> | </a> | ||
− | |||
− | |||
<br /> | <br /> | ||
</a> | </a> | ||
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</h4> | </h4> | ||
<img src="https://static.igem.org/mediawiki/2017/c/c3/NPU-image18.png" style="max-width:60%;"><br /> | <img src="https://static.igem.org/mediawiki/2017/c/c3/NPU-image18.png" style="max-width:60%;"><br /> | ||
− | It can be seen that when the induction temperature was 30 ℃, the enzyme expression and activity were the highest, and the yield of acrylic acid was the best. | + | <h4> |
+ | It can be seen that when the induction temperature was 30 ℃, the enzyme expression and activity were the highest, and the yield of acrylic acid was the best.</h4> | ||
</br> | </br> | ||
+ | <h4> | ||
4.2 the results of production of acrylic acid with different carbon sources | 4.2 the results of production of acrylic acid with different carbon sources | ||
Condition: PH7.4 | Condition: PH7.4 | ||
Reaction time: 16h | Reaction time: 16h | ||
Glucose concentration: 4g/L | Glucose concentration: 4g/L | ||
− | Glycerol concentration: 1%<br /> | + | Glycerol concentration: 1%<br /></h4> |
<img src="https://static.igem.org/mediawiki/2017/8/80/NPU-image19.png" style="max-width:60%;"><br /> | <img src="https://static.igem.org/mediawiki/2017/8/80/NPU-image19.png" style="max-width:60%;"><br /> | ||
+ | <h4> | ||
It can be seen that the yield of acrylic acid was higher when the glycerol was used | It can be seen that the yield of acrylic acid was higher when the glycerol was used | ||
as the carbon source, because the carbon flow rate of the glycerol metabolic | as the carbon source, because the carbon flow rate of the glycerol metabolic | ||
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4.3 The effects of different pH on the amount of acrylic acid were investigated. | 4.3 The effects of different pH on the amount of acrylic acid were investigated. | ||
The results are as follows: | The results are as follows: | ||
− | Reaction conditions: 12h reaction time, 1% concentration of substrate glycerol <br /> | + | Reaction conditions: 12h reaction time, 1% concentration of substrate glycerol </h4><br /> |
<img src="https://static.igem.org/mediawiki/2017/d/d7/NPU-image20.png" style="max-width:60%;"><br /> | <img src="https://static.igem.org/mediawiki/2017/d/d7/NPU-image20.png" style="max-width:60%;"><br /> | ||
− | + | <h4> | |
It can be drawn that PH8.0 was most suitable for acrylic acid production; the | It can be drawn that PH8.0 was most suitable for acrylic acid production; the | ||
reason may be that alkaline environment made E.coli more resistant to acrylic | reason may be that alkaline environment made E.coli more resistant to acrylic | ||
acid. | acid. | ||
4.4 The effect of different Buffer on the amount of acrylic acid were investigated. | 4.4 The effect of different Buffer on the amount of acrylic acid were investigated. | ||
− | The results are as follows: | + | The results are as follows:</h4> |
<img src="https://static.igem.org/mediawiki/2017/8/86/NPU-image21.png" style="max-width:60%;"><br /> | <img src="https://static.igem.org/mediawiki/2017/8/86/NPU-image21.png" style="max-width:60%;"><br /> | ||
− | <img src="https://static.igem.org/mediawiki/2017/e/ef/NPU-image22.png" style="max-width:60%;"><br /> | + | <img src="https://static.igem.org/mediawiki/2017/e/ef/NPU-image22.png" style="max-width:60%;"><br /> <h4> |
It can be seen that the DHa or G3P activity of the two substrates of ceaS2 enzyme was | It can be seen that the DHa or G3P activity of the two substrates of ceaS2 enzyme was | ||
higher under PBS buffer condition. | higher under PBS buffer condition. | ||
− | 4.5 The effects of different reaction time on the amount of acrylic acid were investigated. The results are shown as follows<br /> | + | 4.5 The effects of different reaction time on the amount of acrylic acid were investigated. The results are shown as follows </h4><br /> |
− | <img src="https://static.igem.org/mediawiki/2017/c/c2/NPU-image23.png" style="max-width:60%;"><br /> | + | <img src="https://static.igem.org/mediawiki/2017/c/c2/NPU-image23.png" style="max-width:60%;"><br /> <h4> |
It can be drawn that the yield of acrylic acid reached a higher level after the whole | It can be drawn that the yield of acrylic acid reached a higher level after the whole | ||
cell catalytic reaction endured for 16h. The sampling point should be set after 16h. | cell catalytic reaction endured for 16h. The sampling point should be set after 16h. | ||
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<br /> | <br /> | ||
</h4> | </h4> | ||
− | <h2 id="section-5" style="padding-top: 100px; margin-top: -50px;">5.Conclusion </h2> | + | <h2 id="section-5" style="padding-top: 100px; margin-top: -50px;">5.Conclusion </h2> <h4> |
Due to the time limit of the experiment, we did not have enough time to replace the | Due to the time limit of the experiment, we did not have enough time to replace the | ||
optimal mutation site into the existing cell factory. At present, the highest yield of | optimal mutation site into the existing cell factory. At present, the highest yield of | ||
acrylic acid that we have acquired is 211.655 mg / L, which is 200 times than that of | acrylic acid that we have acquired is 211.655 mg / L, which is 200 times than that of | ||
− | GAACF1.0. | + | GAACF1.0. </h4> |
<br /> | <br /> | ||
<img src="https://static.igem.org/mediawiki/2017/a/a1/NPU-25.png" style="max-width:60%;"><br /> | <img src="https://static.igem.org/mediawiki/2017/a/a1/NPU-25.png" style="max-width:60%;"><br /> |
Latest revision as of 02:22, 2 November 2017