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A method for purifying pea albumin pa1a using negatively charged polysaccharides

A negatively charged, albumin technology, applied in the field of protein extraction, can solve the problems of high cost, high price, low separation speed, etc., and achieve the effects of easy scale, low equipment requirements, and less polysaccharide consumption.

Active Publication Date: 2022-03-15
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the traditional chromatographic column separation has high selectivity, its expensive price and low separation speed make the separation and purification of PA1a have a high cost

Method used

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  • A method for purifying pea albumin pa1a using negatively charged polysaccharides
  • A method for purifying pea albumin pa1a using negatively charged polysaccharides
  • A method for purifying pea albumin pa1a using negatively charged polysaccharides

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] (1) Pretreatment of pea whey: adjust the pH of pea whey to 4.5 with 2 mol / L hydrochloric acid, let it stand at 4°C for 48 hours, centrifuge at 8000rpm for 20 minutes, remove the precipitate, and add 1 mol / L NaOH solution to adjust The pH of the supernatant was adjusted to 7.0 to obtain the pretreated pea whey liquid.

[0026] (2) Negatively charged polysaccharide complex coacervation and precipitation of PA2 and lectin in pea whey protein: Weigh 400 mg of carboxymethylcellulose powder (molecular weight 250 kDa), dissolve it in 100 mL of deionized water, and stir it magnetically for 2 h. It dissolves completely. Add 20% (v / v) carboxymethyl cellulose solution to the pea whey, mix well, adjust the pH to 4.12, stir magnetically for 15 minutes, and centrifuge (4000 rpm, 20 minutes) to remove complex aggregates and precipitates , to collect the supernatant to obtain the crude protein extract of PA1a and carboxymethyl cellulose.

[0027] (3) Purification of PA1a pea albumin:...

Embodiment 2

[0034](1) Pretreatment of pea whey: adjust the pH of pea whey to 4.6, let it stand at 4°C for 24 h, centrifuge at 4000 rpm for 30 min, remove the precipitate, add 1 mol / L NaOH solution to adjust the pH of the supernatant to 7.5, obtaining the pretreated pea whey liquid.

[0035] (2) Negatively charged polysaccharide complex coacervation and precipitation of PA2 and lectin in pea whey protein: Weigh 450 mg of dextran sulfate powder (molecular weight 500 kDa), dissolve it in 100 mL of deionized water, and stir it magnetically for 1 h to make it completely dissolved. Add 10% (v / v) dextran sulfate solution to the pea whey, mix well, adjust the pH to 4.60, stir magnetically for 25 min, and centrifuge (6000 rpm, 15 min) to remove the precipitate of complex aggregates, Collect the supernatant to obtain the crude protein extract of PA1a and dextran sulfate.

[0036] (3) Purification of PA1a pea albumin: adjust the pH of the above solution to 7.5, remove polysaccharides by ultrafiltr...

Embodiment 3

[0041] (1) Pretreatment of pea whey: adjust the pH of pea whey to 4.8, let it stand at 4°C for 48 h, centrifuge at 6000 rpm for 15 min, remove the precipitate, add 1 mol / L NaOH solution to adjust the pH of the supernatant to 7.0, to obtain the pretreated pea whey.

[0042] (2) Negatively charged polysaccharide complex coacervation and precipitation of PA2 and lectin in pea whey protein: Weigh 500 mg of sodium alginate powder (molecular weight 390 kDa), dissolve it in 100 mL of deionized water, and stir it magnetically for 2 h to make it completely dissolve. Add 20% (v / v) sodium alginate solution to the pea whey, mix well, adjust the pH to 3.9, stir magnetically for 30 min, and centrifuge (4000 rpm, 20 min) to remove complex aggregates and precipitates, collect The supernatant is the crude protein extract of PA1a and sodium alginate.

[0043] (3) Purification of PA1a pea albumin: adjust the pH of the above solution to 8.0, remove polysaccharides by ultrafiltration (100 kDa) a...

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Abstract

The invention discloses a method for purifying pea albumin PA1a by using negatively charged polysaccharides, which belongs to the technical field of protein extraction. The invention uses natural edible negatively charged polysaccharides as recovery materials, utilizes the technical principle that the negatively charged polysaccharides can preferentially bind to non-target protein precipitation, and phase-separate from the target protein, realize the separation and purification of pea albumin PA1a, and prepare a single PA1a. From the perspective of resource recovery and utilization, the invention can realize high value-added utilization of pea protein isolate by-products, and prepare high-purity high-sulfur-containing active protein components therefrom, providing technical support for subsequent research. The invention can improve the comprehensive utilization of pea whey, has high recovery rate of PA1a protein and high purity, and has low equipment requirements, simple and quick operation, and no environmental pollution problem.

Description

technical field [0001] The invention relates to a method for purifying pea albumin PA1a by using negatively charged polysaccharides, and belongs to the technical field of protein extraction. Background technique [0002] In recent years, interest in plant-based proteins has grown and people's diets are gradually shifting towards sustainable plant-based ingredients. Among them, pea, as a sustainable crop, is not only a good source of starch products, but also a good source of protein products. As a hypoallergenic protein, pea protein has no risk of genetic modification and has high nutritional value. In European and American countries, pea protein milk drinks and fermented milk have attracted much attention, and have increasingly become important substitutes for milk protein drinks. In my country, the utilization rate of pea protein is not only low, but also the utilization of pea protein is mostly concentrated in pea protein isolate, resulting in the waste of pea whey prot...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C07K1/30C07K1/14
CPCC07K14/415
Inventor 李兴飞华欲飞杨淑暖陈业明孔祥珍张彩猛
Owner JIANGNAN UNIV
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