Protaetia brevitarsis larva protein PB3, separation method thereof and application of protaetia brevitarsis larva protein PB3
A kind of white star flower beetle, protein technology, applied in the field of agricultural biology, can solve the problem of rare insect protein and so on
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Embodiment 1
[0047] Example 1 Extraction of the anti-inflammatory protein of the larvae of the white star beetle
[0048] 1. Extraction
[0049] Salt extraction method: Accurately weigh 50.00g of defatted Beetle larvae sample, add 750mL of 1.5% NaCl solution at a ratio of 1:15 (g / mL), stir in a water bath for 1.5h at a constant temperature of 55°C, and rotate at a speed of 8000r / min Centrifuge for 10 min and take the supernatant. Slowly pour 8.00 mol / L hydrochloric acid solution into the supernatant until no more precipitation occurs, centrifuge at 8000r / min for 10min, take the precipitate, and freeze-dry it for later use.
[0050] Acid extraction method: Accurately weigh 50.00g of defatted Beetle larvae sample, add 750mL of 0.1mol / L acetic acid solution at a ratio of 1:15 (g / mL), stir in a water bath for 1.5h at a constant temperature of 55°C, and rotate at 8000r / min centrifuged for 10 min, the supernatant was taken, and the precipitate was repeatedly extracted once. The supernatant wa...
Embodiment 2
[0063] Example 2 Optimization of Extraction Conditions for the Anti-inflammatory Protein of Beetle larvae
[0064] Taking the protein extraction rate and total anti-inflammatory ability of the larvae of the white star beetle as the measurement index, the NaOH concentration (g / mL), solid-liquid ratio (defatted insect powder: NaOH solution = M:V), hydrolysis time (h), and precipitation were investigated. Effect of pH and hydrolysis temperature (°C) on metrics.
[0065] (1) Condition optimization of solid-liquid ratio (defatted insect powder: NaOH solution=M:V)
[0066] Under the condition of 55°C, the extraction time was 1.5h, the NaOH concentration in the extraction system was 1.5% (g / mL), the pH value of the precipitation was 4, and the solid-liquid ratio (defatted insect powder: NaOH solution = M:V) was 1:5, 1 Under the conditions of :10, 1:15, 1:20, and 1:25, measure each measurement index.
[0067] like Figure 8 It was shown that when the ratio of the mass of defatted i...
Embodiment 3
[0080] Example 3 Detection of anti-inflammatory effect of anti-inflammatory protein of white star beetle larvae
[0081] Take out the freeze-dried sample components, weigh an appropriate amount and dissolve them in distilled water. Take out the dialysis bag, soak it in distilled water and wash it, inject 5mL of sample solution into the dialysis bag, clamp and seal both ends of the dialysis bag, put it into a beaker filled with distilled water, place it on a rotator, change the water every 20 minutes, and detect it with silver nitrate. Until no white precipitate appears in the beaker, it proves that the dialysis is complete. Pour out the sample solution in the dialysis bag, and freeze dry it for storage.
[0082] 1. The effect of anti-inflammatory protein from the larvae of the white star beetle on the production of NO in RAW264.7 cells induced by LPS
[0083] After optimizing the extraction conditions, the larval anti-inflammatory proteins were extracted. The extracted crud...
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