Difference between revisions of "Part:BBa K4146507"
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− | + | We hope to use 2-PE as an example to give E. coli Nissle 1917 the ability to sweeten hair in addition to improving scalp microecology. We use the natural phenylalanine synthesis pathway of E. coli to synthesize 2-PE. aroG was used to enhance the accumulation of a key intermediate, DAHP. aroG, as a metabolic pathway gene that has attracted much attention for many years, has always been a worthy object of study. We believe that its feedback inhibition may affect the synthesis of 2-phenylethyl alcohol. We use this Mutant Feedback-Resistant Version of aroG for to resisting feedback inhibition and improving enzyme activity. This is the composite of our Improvement of Standard Biological Parts. | |
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[[File:The standard curve for determination of 2-phenylethanol yield.png|500px]] | [[File:The standard curve for determination of 2-phenylethanol yield.png|500px]] | ||
− | The standard curve for determination of 2-phenylethanol yield | + | Fig.1 The standard curve for determination of 2-phenylethanol yield |
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+ | In the standard curve, as shown in Fig.1,the ordinate is the peak area and the abscissa is the marker concentration.The mobile phase of high performance liquid chromatography is trifluoroacetic acid diluted 1000 times.In our experiment, we took two points, 500 mg/L and 1000 mg/L, as well as zero point. The R square fitted by the standard curve is 1, which shows that the experimental results are fitted very well, and the error can be almost ignored, that is, the measured output is relatively accurate. | ||
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[[File:The experimental 2-phenylethanol yield.png|500px]] | [[File:The experimental 2-phenylethanol yield.png|500px]] | ||
− | The experimental 2-phenylethanol yield | + | Fig.2 The experimental 2-phenylethanol yield |
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+ | Our team successfully constructed a pathway for the synthesis of 2-phenylethanol in Escherichia coli. When cultured for 60 hours, the maximum concentration of 2-phenylethanol was 67.16mg/L.After 60 hours,the concentration of 2-phenylethanol decreases because of the toxicity of 2-PE accumulated in E. coli and metabolism. | ||
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Latest revision as of 15:31, 11 October 2022
pCS-aroGfbr
We hope to use 2-PE as an example to give E. coli Nissle 1917 the ability to sweeten hair in addition to improving scalp microecology. We use the natural phenylalanine synthesis pathway of E. coli to synthesize 2-PE. aroG was used to enhance the accumulation of a key intermediate, DAHP. aroG, as a metabolic pathway gene that has attracted much attention for many years, has always been a worthy object of study. We believe that its feedback inhibition may affect the synthesis of 2-phenylethyl alcohol. We use this Mutant Feedback-Resistant Version of aroG for to resisting feedback inhibition and improving enzyme activity. This is the composite of our Improvement of Standard Biological Parts.
Fig.1 The standard curve for determination of 2-phenylethanol yield
In the standard curve, as shown in Fig.1,the ordinate is the peak area and the abscissa is the marker concentration.The mobile phase of high performance liquid chromatography is trifluoroacetic acid diluted 1000 times.In our experiment, we took two points, 500 mg/L and 1000 mg/L, as well as zero point. The R square fitted by the standard curve is 1, which shows that the experimental results are fitted very well, and the error can be almost ignored, that is, the measured output is relatively accurate.
Fig.2 The experimental 2-phenylethanol yield
Our team successfully constructed a pathway for the synthesis of 2-phenylethanol in Escherichia coli. When cultured for 60 hours, the maximum concentration of 2-phenylethanol was 67.16mg/L.After 60 hours,the concentration of 2-phenylethanol decreases because of the toxicity of 2-PE accumulated in E. coli and metabolism.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 450
Illegal EcoRI site found at 665
Illegal XbaI site found at 87
Illegal XbaI site found at 941 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 450
Illegal EcoRI site found at 665 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 450
Illegal EcoRI site found at 665
Illegal BamHI site found at 738 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 450
Illegal EcoRI site found at 665
Illegal XbaI site found at 87
Illegal XbaI site found at 941 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 450
Illegal EcoRI site found at 665
Illegal XbaI site found at 87
Illegal XbaI site found at 941 - 1000COMPATIBLE WITH RFC[1000]