Application of enzymolysis cereal bran extract to preparation of DPP-4 inhibitor
An extract and bran technology, applied in the biological field, can solve problems such as discarding and waste of resources, and achieve the effects of good quality, resource saving and production cost reduction
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Embodiment 1
[0046] (1) Drying: Weigh 50g of oat bran, dry in an oven at 40°C for 24 hours, and pass through an 80-mesh sieve after crushing;
[0047] (2) Enzymolysis: Mix the pulverized bran and water according to the ratio of material to liquid: 1g:10mL, add cellulase, mix evenly, stir the enzymolysis with a magnetic stirrer at 300rpm in a water bath at 50°C for 3h;
[0048] (3) Extraction: Add the ethanol-water mixture with a volume fraction of 85% to the enzymolysis solution according to the solid-liquid ratio of the grain bran and the extraction solution at 1g:30mL, and extract at 40°C and 360W ultrasonic conditions for 1 hour to obtain the extract , centrifuge the extract at 4000rpm for 10 minutes to obtain the supernatant and precipitate, collect the supernatant; add the extract solution to the precipitate, prepare the extract under ultrasonic conditions, and centrifuge the extract at 4000rpm for 10 minutes to obtain the supernatant and precipitate, collect supernatant; add extracti...
experiment example 1
[0111] Experimental Example 1 Analysis of the efficacy and composition of enzymatically hydrolyzed cereal bran extract
[0112] 1. High performance liquid chromatography
[0113] The concentrated extract was filtered through a 0.22 μm microporous membrane for HPLC mass spectrometry analysis, using Inertsil ODS-3C 18 (150mm*4.6mm*3μm) chromatographic column completes the separation. The composition of the mobile phase is: A 5% formic acid aqueous solution; B 95% methanol and 5% acetonitrile mixed solution. The gradient elution conditions are: 0-18min, 0-80%B; 18-20min, 80-100%B; 20-21min, 100%B; 21-22min, 100-0%B. The injection volume was 10 μL, the flow rate was 0.5 mL / min, and the total run time was 22 min, with monitoring at 280 nm, 360 nm and 520 nm. Under the same conditions, the composition of flavonoids was analyzed compared with the retention time of the standard.
[0114] Such as figure 1 As shown, there are mainly 9 kinds of flavonoids in the DPP-4 inhibitor in t...
experiment example 2
[0118] Determination of the DPP-4 inhibitory activity of experimental example 2 cereal by-product extract
[0119]25 μL of an appropriate concentration of the sample solution to be tested and 50 μL of 3mM GPPN solution (0.1M, pH8.0) (luminescent substrate Gly-Pro-pNA, Sigma Chemical Co) were shaken and mixed, then incubated at 37°C for 10 min, and 50 μL of 16U / LDPP-4 solution (Sigma Chemical Co) to start the reaction, vortexed and mixed, and the mixture was incubated at 37°C for 90 min, then 100 μL of sodium acetate buffer (0.1M, pH 4.0) was added to terminate the reaction, vortexed and mixed in Absorbance was measured at 450nm.
[0120] DPP-4 inhibition rate = [1-(A s -A b ) / (A c -A 0 )]*100%
[0121] Among them, A c is the absorbance value of the reaction solution without adding the sample, A 0 For the enzyme solution blank, A s is the absorbance value of the reaction solution added with the sample, A b blank for the sample.
[0122] image 3 It is the DPP-4 inhi...
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