Extraction method for antioxidant polypeptide in Guangchang nymphaea alba
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An antioxidant polypeptide and extraction method technology, applied in the field of antioxidant polypeptide extraction, can solve problems such as complex biosafety purification process, and achieve the effects of strong antioxidant capacity, simple process, and controllable performance.
Active Publication Date: 2013-04-03
瑞金市工业投资发展有限公司
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[0005] The purpose of the present invention is to provide a method for obtaining white lotus protein polypeptides with high antioxidant properties in view of the two problems existing in the extraction of white lotus protein polypeptides
It can effectively solve the two problems of biological safety and the complexity of the subsequent purification process
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Embodiment 1
[0015] The protein extraction of Guangchang white lotus adopts the following steps.
[0016] (1) Grind the white lotus into powder with a mixing machine, sieve out about 99% of the fine powder with a single layer of gauze, and sift out about 85% of the fine powder with a double layer of gauze. Put the white lotus powder into water at 60°C, the water contains about 3% hydrogen peroxide, and the ratio of white lotus powder to water is 1:12.
[0017] (2) Add sodium hydroxide aqueous solution, and keep stirring until the pH value is 8-9, let it stand for 30 minutes, remove the oily and milky substances, and separate the solid and liquid to obtain solid and liquid substances.
[0018] (3) Mix and stir the solid and water in step (2) at a ratio of about 5:1, and separate the water according to step (2), mix the water with the liquid in (2), and wash with hydrochloric acid The pH was adjusted to about 4.5, and the protein solids were precipitated. Centrifuge for 15 minutes to obtai...
Embodiment 2
[0021] Weigh 15g of Guangchang white lotus protein and prepare it into an aqueous solution with a substrate concentration of 5%. After dissolving, the solution is quickly heated to 85°C and kept at a constant temperature for 15min. After pretreatment, add phosphate buffer to adjust the pH to 8.5, add trypsin, enzymolysis temperature is 45°C, enzyme dosage is 9% of the substrate concentration, after enzymolysis for 120min, the enzymolysis solution is rapidly heated to 90°C, Keep it for 20min, and carry out the enzyme-inactivating treatment. Then rapidly cool to room temperature, centrifuge at 3000r / min for 20min, and take the supernatant. At this time, the degree of hydrolysis of white lotus protein is 37.25%. According to the ratio of 100 grams: 100ml, trypsin hydrolyzate was added to the wet macroporous adsorption resin DA201-C, kept on a constant temperature shaker at 25°C for 180min, the rotation speed was 150r / min, and eluted with 75% ethanol solution for 120min, collected...
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Abstract
An extraction method for antioxidant polypeptide in Guangchang nymphaea alba is characterized by comprising the following steps of weighting a quantity of protein of Guangchang nymphaea alba for preparing water solution with substrate concentration of 3-6%, heating the solution rapidly to 80-85 DEG C after dissolution, and keeping the temperature for 10-15 minutes; after pretreatment, adding phosphate buffer so as to adjust the pH to 7.0-8.5, adding trypsin, wherein the enzymolysis temperature is 45-60 DEG C, and the enzyme use amount is 6-9% of substrate concentration, after 60-120 minutes of enzymolysis, rapidly heating enzymatic hydrolysate to 90 DEG C for 20 minutes for enzyme deactivation treatment; then rapidly cooling down to room temperature, carrying out centrifugation by 3000-4000r / min for 15-20 minutes, taking supernate, adding trypsin solution into wet macroreticular resin DA201-C according to the proportion of 100g:100-120ml, keeping in a constant temperature table for 180-240 minutes, wherein the temperature is 25 DEG C and the revolving speed is 150-200r / min, eluting sequentially with ethanol solution of 25%, 50%, 75% and 100% for 120-150 minutes, and collecting eluant for vacuum drying and concentration. The method is simple and is controllable, an HPLC-MS analysis result shows that polypeptide molecular weight focus between 1100-1900D, and the oxidation resistance is stronger than polypeptide obtained through microbial fermentation.
Description
technical field [0001] The invention belongs to the extraction of peptides in natural resources, in particular to the extraction of antioxidant polypeptides. technical background [0002] Studies have shown that the accumulation of free radicals is related to the occurrence of many diseases in humans and other animals, such as cancer, aging, arteriosclerosis, diabetes, fatigue, low immunity, and sub-health. Proper intake of substances with antioxidant activity can reduce the level of free radicals in the body and help the body resist diseases caused by the accumulation of free radicals. Exogenous antioxidants can come from chemical synthesis and natural extraction. The use of chemically synthesized antioxidants is increasingly limited due to potential health hazards. [0003] In recent years, people have turned their attention to natural and safe antioxidants from animals and plants. Natural antioxidants can not only prevent the damage of free radicals to the body, but al...
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