Polyoxygenated cyclohexene derivatives and application thereof
A cyclohexene and oxygen substitution technology, applied in the field of microorganisms, can solve problems such as cyclohexene compounds that have not yet been seen
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Embodiment 1
[0024] Separation and preparation of polyoxygen substituted cyclohexene compounds A-G:
[0025] 1. Slant culture of Phomopsis YE3250 strain: peel potatoes, cut into small pieces, add water to boil, filter with gauze to obtain potato filtrate, then add glucose and agar, constant volume with distilled water, and sterilize to obtain slant medium; The medium is made into a test tube slant, and the YE3250 strain is picked and inserted into the slant medium for cultivation to obtain the strain slant;
[0026] 2. Seed culture of Phomopsis YE3250 strain: Take a strain transplant block from the above slant strains and inoculate it into PDB medium (PDB medium: 200 g of peeled potatoes, 20 g of glucose, 15 g of agar, 1000 g of distilled water mL, natural pH, sterilized at 121°C for 30 minutes), cultured on a shaker at 28°C and 200 r / min for 4 days to obtain seed solution;
[0027] 3. Liquid fermentation culture of Phomopsis YE3250 strain: 200 mL of PDB medium was placed in a 500 mL E...
Embodiment 2
[0030] The inhibitory activity of the polyoxo-substituted cyclohexene compounds A-G of the present invention on a-glucosidase was detected in a 96-well plate by using a microplate reader. The samples to be tested were dissolved in phosphate buffer (PBS, pH 6.8) at three concentrations. Add 20 μL of 0.01M phosphate buffer and 20 μL of phosphate buffer of 2.5 mM substrate 4-nitrophenol-α-D glucopyranoside (PNPG) into the 96-well plate in advance, then add 10 μL of sample solutions of different concentrations, Incubate for 5 minutes at 37°C in a standard instrument. Then add 10 μL of 0.2 U / mL α-glucosidase solution prepared with 0.01M phosphate buffer (pH 6.8) to start the reaction, and react at 37°C for 15 minutes, then add 0.2M Na 2 CO 3 80 μL was used to stop the reaction, and the absorbance OD value was measured at 405 nm. Acarbose was used as the positive control, PBS was used instead of α-glucosidase in the blank group, and the negative control was 20 μL PBS+20 μL PNPG+...
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