Method for separating and purifying soybean agglutinin from soybean whey

A technology for soybean lectin and soybean whey, which is applied in the preparation methods of peptides, chemical instruments and methods, organic chemistry, etc., can solve the problems of low purification efficiency and purity of guar gum, high cost, limited large-scale use, and cumbersome and complicated procedures. and other problems, to achieve the effect of low equipment requirements, high recovery rate and high purity

Active Publication Date: 2015-04-22
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are certain limitations in the above systems. The purification efficiency and purity of guar gum are low. N-acetyl-D-galactosamine-Sepharose 6B is limited by the high cost of ligand N-acetyl-D-galactosamine. For large-scale use, the flow rate of galactosamine-CH-Sepharose4B is limited, and the above separation and purification procedures have relatively cumbersome and complicated procedures, and the condition control is relatively strict

Method used

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  • Method for separating and purifying soybean agglutinin from soybean whey
  • Method for separating and purifying soybean agglutinin from soybean whey
  • Method for separating and purifying soybean agglutinin from soybean whey

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Adjust the soybean whey to pH 4.5, centrifuge at 9500rpm for 30min to remove the precipitate; adjust to pH 8.0 again, centrifuge at 9500rpm for 30min, discard the precipitate, and collect the supernatant. Adjust the pH of the above supernatant to 4.5, measure 1L of the solution, add solid ammonium sulfate to 50% saturation according to the ammonium sulfate saturation table at 4-30°C, centrifuge at 9500rpm for 30min, collect the precipitate, and continue to add sulfuric acid to the supernatant Ammonium to 60% saturation, centrifuged again at 9500rpm for 30min, collected the precipitate, combined the two precipitates to obtain about 0.65g of the crude soybean lectin protein contained in it, dissolved it in 10mL deionized water, dialyzed and desalted it for 36 hours with a molecular weight cut-off of 3500, and vacuumed the retentate Freeze-dry to prepare a P-7S protein sample containing soybean lectin and β-amylase.

[0021] Accurately weigh 0.1g of the above primary isola...

Embodiment 2

[0025] Adjust the soybean whey to pH 4.7, centrifuge at 9500rpm for 30min to remove the precipitate; adjust to pH 8.5 again, centrifuge at 9500rpm for 30min, discard the precipitate, and collect the supernatant. Adjust the pH of the above supernatant to 4.5, measure 2L of the solution, add solid ammonium sulfate to 50% saturation according to the ammonium sulfate saturation table at 4-30°C, centrifuge at 9500rpm for 30min, collect the precipitate, and continue to add sulfuric acid to the supernatant Ammonium to 70% saturation, centrifuged again at 9500rpm for 30min, collected the precipitate, combined the two precipitates to obtain about 1.85g of crude soybean lectin protein, dissolved in 30mL of deionized water, dialyzed and desalted for 48 hours with a molecular weight cut-off of 3500, and vacuumed the retentate Freeze-dry to prepare a P-7S protein sample containing soybean lectin and β-amylase.

[0026] Accurately weigh 0.3g of the above-mentioned primary isolated P-7S prot...

Embodiment 3

[0030] Adjust the soybean whey to pH 4.8, centrifuge at 9500rpm for 30min to remove the precipitate; adjust to pH 9.0 again, centrifuge at 9500rpm for 30min, discard the precipitate, and collect the supernatant. Adjust the pH of the above supernatant to 4.7, measure 1.5L of the solution, add solid ammonium sulfate to 50% saturation according to the ammonium sulfate saturation table at 4-30°C, centrifuge at 9500rpm for 30min, collect the precipitate, and continue to add the supernatant Ammonium sulfate to 80% saturation, centrifuged again at 9500rpm for 30min, collected the precipitate, combined the two precipitates to obtain about 1.57g of crude soybean lectin protein, dissolved in 30mL of deionized water, dialyzed and desalted for 48 hours with a molecular weight cut-off of 3500, and vacuum freeze-dried , to prepare a P-7S protein sample containing soybean lectin and β-amylase.

[0031] Accurately weigh 0.4g of the above-mentioned primary isolated P-7S protein sample and 0.4g...

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Abstract

The invention relates to a method for separating and purifying soybean agglutinin from soybean whey and belongs to the field of processing of agricultural products and comprehensive utilization of byproducts. The method disclosed by the invention comprises the following steps: (1) pretreating soybean whey; (2) primarily separating soybean whey protein; (3) re-agglomerating the soybean whey protein and poly-anions; and (4) recycling the soybean agglutinin. Raw materials are pretreated, primarily separated and then re-agglomerated to obtain a final protein polysaccharide compound; the final protein polysaccharide compound is ultra-filtered to remove sugar and then is secondarily centrifuged to obtain a protein solution containing purified soybean agglutinin; the protein solution is subjected to vacuum freeze-drying to obtain a high-purity sample of the soybean agglutinin. The method disclosed by the invention can be used for comprehensively utilizing the soybean whey and is low in requirements on equipment, simple to operate and free of environmental pollution; the soybean agglutinin is high in recycle rate, high in purity and high in protein activity and still retains natural physiological activity.

Description

technical field [0001] The invention relates to a method for separating and purifying soybean lectin from soybean whey, specifically a new method for recovering soybean lectin in soybean whey by using polyanionic polysaccharides, and belongs to the field of agricultural product processing and comprehensive utilization of by-products. Background technique [0002] Soy whey protein is the protein that remains in soybean whey and cannot be precipitated by acid. In soybean whey protein, the 2S component accounts for a large proportion; whey protein not only contains globulin and albumin, but also Mainly contains: ①Kunitz trypsin inhibitor (KTI, pH3.0~10.0, 20kDa); ②Bowman-Brik trypsin inhibitor (BBI, pH3.0~10.0, 20KDa); ③β-amylase (61.7KDa); ④Agglutination (pH 2.2~10.8, 120KDa); ⑤lipoxygenase (LOX, 102KDa) and other physiologically active substances, accounting for about 9%-15.3% of soybean protein. [0003] As one of the main anti-nutritional factors in soybean, soybean lectin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/42C07K1/34
CPCC07K14/42
Inventor 华欲飞李兴飞孔祥珍张彩猛陈业明
Owner JIANGNAN UNIV
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