Difference between revisions of "Part:BBa K4348000"

(Introduction)
(Introduction)
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==Results==
 
==Results==
[[File:AKR1D1 popassay results.png|200px|thumb|left|<strong>Figure 3. GC-MS chromatogram of three AKR1D1 pop assays in E. coli mixed with cholestenone substrate.</strong> The GC-MS chromatogram shows the 0.01% coprostanone standard along with the product of a three AKR1D1 reactions containing, 100µM cholestenone, 500µM NADPH, 100mM potassium phosphate buffer (pH 6.0), 0.2% Triton X-100, and 5% ethanol, incubated for 16 hours at 37°C. AKR1D1 was added to the reaction using the pop assay protocol. The AKR1D1 reaction chromatogram shows a peak directly underneath the coprostanone standard, indicating product formation using the pop assay protocol.]]
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[[File:AKR1D1 popassay results.png|400px|thumb|left|<strong>Figure 3. GC-MS chromatogram of three AKR1D1 pop assays in E. coli mixed with cholestenone substrate.</strong> The GC-MS chromatogram shows the 0.01% coprostanone standard along with the product of a three AKR1D1 reactions containing, 100µM cholestenone, 500µM NADPH, 100mM potassium phosphate buffer (pH 6.0), 0.2% Triton X-100, and 5% ethanol, incubated for 16 hours at 37°C. AKR1D1 was added to the reaction using the pop assay protocol. The AKR1D1 reaction chromatogram shows a peak directly underneath the coprostanone standard, indicating product formation using the pop assay protocol.]]
  
 
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Revision as of 19:44, 13 October 2022


AKR1D1_his

A human 5-beta reductase used in bile acid synthesis. Used in the 2022 McGill project to catalyze the second step of the cholesterol -> coprostanol pathway, which is 4-cholesten-3-one to coprostanone. Coprostanol cannot be absorbed by the gut, which is a unique property.

Introduction

yoyoyo

Biology

Results

Figure 3. GC-MS chromatogram of three AKR1D1 pop assays in E. coli mixed with cholestenone substrate. The GC-MS chromatogram shows the 0.01% coprostanone standard along with the product of a three AKR1D1 reactions containing, 100µM cholestenone, 500µM NADPH, 100mM potassium phosphate buffer (pH 6.0), 0.2% Triton X-100, and 5% ethanol, incubated for 16 hours at 37°C. AKR1D1 was added to the reaction using the pop assay protocol. The AKR1D1 reaction chromatogram shows a peak directly underneath the coprostanone standard, indicating product formation using the pop assay protocol.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]