Chinese yam protein extract with pruritus-causing activity as well as preparation method and application thereof
A protein extract, yam technology, applied in the field of biomedicine, to achieve the effects of not easy to store, good application prospects, and high scientific research value
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Embodiment 1
[0026] Example 1 Preparation method of yam protein component with pruritic activity and its pruritus evaluation.
[0027] A preparation method of yam protein extract with pruritic activity specifically comprises the following steps.
[0028] 1. Sample pretreatment.
[0029] Wash and slice the yam, freeze at -20°C for 12 hours, take 500g of yam sample and thaw it, add 1L of purified water, homogenize for 20min to obtain a solid-liquid mixture, centrifuge at 11500rpm at 4°C for 25min, take the supernatant, and measure the pH of the supernatant to be 6.35 , and the conductivity was 3.0 mS / cm, and the sample solution 1 was obtained.
[0030] Concentrate sample solution 1 with a 3kDa ultrafiltration membrane to a volume of about 200ml, adjust the pH to 8.0 with NaOH, and perform membrane filtration in the order of 0.8μm+0.45μm to obtain sample solution 2.
[0031] 2. Purification process.
[0032] Wash the Q-HPR prepacked column with 20mM Tris, pH 8.0 solution (solution A), wash...
Embodiment 2
[0037] Example 2 Component analysis of pruritic protein.
[0038] The yam protein component with itching activity prepared in Example 1 was analyzed by liquid chromatography-mass spectrometry, and the peptide was dissolved with liquid chromatography mobile phase A (aqueous solution of 0.1% formic acid and 2% acetonitrile), EASY-nLC 1000 ultra-high performance liquid phase system was used for separation. Liquid phase gradient setting: 0-90min, 6%~22%B; 90-112min, 22%~35%B; 112-116min, 35%~80%B; 116-120min, 80%B. The flow rate was set at 400 nL / min. Peptides were separated by an ultra-high performance liquid phase system and injected into an NSI ion source for ionization, and then analyzed by an Orbitrap Fusion mass spectrometer. The data acquisition mode uses the Data Dependent Scan (DDA) procedure.
[0039]Database Search MS / MS data were searched using Maxquant (v1.5.2.8). Search parameter setting: the database is Dioscorea_oppositifolia_TX (44669 sequences), the anti-base...
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