A method for detecting benzyl alcohol acetyltransferase activity
A technology of acetyltransferase and benzyl alcohol, which is applied in the field of biochemistry, can solve the problems of unfavorable development and research of benzyl alcohol acetyltransferase, cumbersome operation steps, and long experiment cycle, so as to eliminate the influence, simple experimental operation steps and high detection results Sensitive and accurate effect
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Embodiment 1
[0026] Example 1 Analysis of Benzyl Alcohol Acetyltransferase Activity in Crude Extracted Protein of Plum Blossom Petals
[0027] This embodiment provides a method for detecting the activity of benzyl alcohol acetyltransferase in crude protein extracted from plum blossom petals, including:
[0028] 1. Mother liquor configuration of experimental reagents:
[0029] (1) Preparation of 10mM DTNB mother solution:
[0030] Weigh 0.2g of DTNB powder and dissolve it in 50mL 0.5M Tris-HCl (pH8.0), store in the dark, and prepare immediately after use;
[0031] (2) Preparation of 50mM acetyl-CoA mother solution:
[0032] Add 50mg of acetyl-CoA powder to 1.2mL of water, mix until complete dissolution, and store at -20°C;
[0033] (3) Preparation of 1M benzyl alcohol mother liquor:
[0034] Add 100 μL benzyl alcohol (analytical grade) to 864 μL DMSO, mix well, and store at -20°C;
[0035] (4) 1mM MgCl 2 Preparation of mother liquor:
[0036] 1M MgCl 2 Obtained by diluting the mothe...
Embodiment 2
[0058] Example 2 Analysis of Benzyl Alcohol Acetyltransferase Activity of Tobacco Leaf Crude Extract Protein
[0059] This embodiment provides a detection of the activity of benzyl alcohol acetyltransferase of the crude extract protein of tobacco leaves, including:
[0060] 1, experimental reagent mother solution configuration (same as embodiment 1)
[0061] 2. Configuration of protein benzyl alcohol acetyltransferase activity detection system:
[0062] (1) Experimental group:
[0063] Add 15 μL of DTNB mother solution, 15 μL of acetyl-CoA mother solution, 1.5 μL of benzyl alcohol mother solution, and MgCl in a 2 mL plastic centrifuge tube. 2 15 μL of the mother liquor and 200 μg of the crude tobacco leaf protein sample were made up to 1.5 mL with water, and mixed. The samples in the experimental group were repeated three times.
[0064] (2) Control group:
[0065] Boil 200 μg of crude tobacco leaf protein sample in boiling water for 10 minutes, centrifuge at 12,000 rpm f...
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