Streptomyces lavendulae and method and application thereof for preparing alpha-glucosidase inhibitor
A technology of Streptomyces lilacinus and glucosidase is applied in the preparation of α-glucosidase inhibitor, the application field of α-glucosidase inhibitor in the treatment of diabetes, and the formation of postprandial hyperglycemia can be improved and reduced. Effects of postprandial blood sugar levels
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Embodiment 1
[0047] Embodiment 1: Isolation and screening of Streptomyces lilacinus gray Streptomyces UN-8
[0048] (1) Preparation of bacterial suspension: Weigh 5 g of soil samples into a 250 mL Erlenmeyer flask filled with 45 mL of sterile saline, and stir magnetically for 30 minutes at room temperature to prepare bacterial suspension.
[0049] (2) Bacterial suspension dilution and colony picking: The bacterial suspension obtained above was diluted into bacterial suspensions of different concentrations using the 10-fold dilution method, and 10 4 , 10 5 , 10 6 100 μL of the bacterial suspension was spread on the solid separation medium plate; after culturing at 28°C for 5 days, single colonies were picked according to the colony shape and size of the actinomycetes and the color of the pigment produced.
[0050] (3) Streak purification: use a flat and smooth inoculation loop to continuously streak and purify the obtained single colony on the Gaoshi No. 1 solid medium plate under asept...
Embodiment 2
[0058] Embodiment 2: the culture characteristic of streptomyces lilacinus griseus UN-8
[0059] Streptomyces lilacinus griseus UN-8 was inoculated in a 250mL Erlenmeyer flask equipped with 50mL carbon-free liquid medium, and the liquid medium added with 2.5g / L and 10g / L glucose was used as a control group respectively, and the shaking table speed was 160rpm. After culturing at 28°C for 7 hours, the growth of the strain was judged by the dry weight method or turbidimetric method. The results showed that the growth of Streptomyces lilacaceae UN-8 in the carbon-free liquid medium and 2.5g / L glucose liquid medium was good. And it is better than the growth of Streptomyces lilacinus UN-8 in 10g / L glucose liquid medium.
[0060] Inoculate Streptomyces lavenderus UN-8 in a 250mL Erlenmeyer flask filled with 50mL nitrogen-free liquid medium, add 1g / L yeast extract and 1g / LKNO 3 The liquid culture medium was used as the control group, the shaker speed was 160rpm, and after culturing ...
Embodiment 3
[0063] Embodiment 3: the method for preparing α-glucosidase inhibitor by Streptomyces lilacaceae UN-8
[0064] (1) Fermentation: Use a flat and smooth inoculation loop to pick a lawn with a length of 3 cm on the inclined surface and insert it into a 500 mL conical flask containing 100 mL of liquid seed medium. Put 10% of the inoculum into a 3L Erlenmeyer flask containing 800mL of liquid fermentation medium, shake the table at a speed of 160rpm, cultivate at 30°C for 3 days, collect the fermentation broth of 5 Erlenmeyer flasks, about 4L, and use it for α-glucosidase Separation and extraction of inhibitors;
[0065] (2) Pretreatment of fermentation broth: Centrifuge the fermentation broth at 8000rpm and 4°C for 15 minutes to remove bacteria and insoluble particles, then filter the supernatant through an ultrafiltration system with a molecular weight cut-off of 5000Da, and the average membrane flux is 30LMH, remove the residual soluble starch, protein and part of the pigment ...
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