Alpha-lactalbumin small-molecular-weight hydrolysate, preparation method thereof and application of alpha-lactalbumin small-molecular-weight hydrolysate in reducing uric acid
A lactalbumin and low molecular weight technology is applied in the field of preparation of low molecular weight hydrolyzates of alpha-lactalbumin, which can solve problems such as large side effects, and achieve the effects of reducing uric acid kidney damage, reducing kidney damage, and having good application prospects.
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Embodiment 1
[0035] Embodiment 1, preparation of α-lactalbumin hydrolyzate F5 component
[0036] α-lactalbumin was dissolved in deionized water to prepare a 40mg / mL solution, adjusted to pH 1.5-2.5 with 5M HCl, added pepsin (250U / mg), and the ratio of pepsin to substrate was 1:30 (w / w), enzymatic hydrolysis at 35-40°C and 150-200rpm for 2-4 hours; then adjust the pH of the system to 6.5-7.5 with 6M NaOH solution, add trypsin (8×USP), trypsin and base The material ratio is 1:25 (w / w), and the hydrolysis is carried out for 3-5 hours at a temperature of 35-40°C and a shaker speed of 150-200 rpm.
[0037]After hydrolysis, adjust the pH of the system to 6.5-7.5, and inactivate in a water bath at 90-100°C. Then the hydrolyzate was centrifuged at 12000 rpm for 10 min, and the supernatant was ultrafiltered with an ultrafiltration tube (50 mL) with a filter diameter of 3 kDa, and the filtrate was collected, which was a hydrolyzate <3 kDa.
Embodiment 2
[0040] Embodiment 2, the determination of XO inhibitory activity in vitro
[0041] Take the complete α-lactalbumin hydrolyzate, the filtrate obtained by ultrafiltration (hydrolyzate <3kDa) and the F5 fraction (molecular weight <1500Da), and measure the inhibitory activity at a concentration of 2 mg / mL. The results are shown in Table 1 .
[0042] Table 1. XO inhibitory activity of different samples
[0043]
[0044] From the results in Table 1, it can be seen that at a concentration of 2mg / mL, the F5 component can inhibit 90% of the XO activity, which is significantly better than that of α-lactalbumin hydrolyzate and <3kDa hydrolyzate.
Embodiment 3
[0045] The determination of the uric acid-lowering effect of embodiment 3, F5 component to hyperuricemia mouse model
[0046] The experimental modeling uses clean-grade, six-week-old ICR male mice. All experimental animals have free access to food and water, the environment is 24±2°C, and the light is 12 hours a day. The modeling method for hyperuricemia was 250 mg / kg urate oxidase inhibitor oxonate potassium and 300 mg / kg hypoxanthine by intragastric administration every day for 3 weeks. The experiments were divided into normal group, model group, allopurinol group (positive control), <3kDa hydrolyzate group and F5 group. Except the normal group, other groups were modeled with high uric acid. The specific dosage is as follows:
[0047] 1) Normal group: intragastric administration of 0.5% sodium carboxymethylcellulose solution of the same volume every day during the experimental period;
[0048] 2) Model group: intragastric administration of 0.5% sodium carboxymethylcellulo...
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