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Method for extracting beta-conglutin from Australian sweet lupin meal

A technology of lupin and detection method, which is applied in the preparation methods of peptides, chemical instruments and methods, plant peptides, etc., can solve the problems of nutritional value impact, protein denaturation, etc., and achieve the effect of preventing denaturation and high purity

Inactive Publication Date: 2019-09-20
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Alkali-soluble acid precipitation is the most common method adopted in industry at present, but this method may cause protein denaturation, which will have a certain impact on nutritional value
And there is currently no mature method for extracting pure lupine β-conglutin. Therefore, a mature method for extracting pure lupine β-conglutin protein is established, and it will be used for further exploration of protein properties, production of corresponding specific antibodies, and human toxicity. It is important to lay the groundwork for research

Method used

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  • Method for extracting beta-conglutin from Australian sweet lupin meal
  • Method for extracting beta-conglutin from Australian sweet lupin meal
  • Method for extracting beta-conglutin from Australian sweet lupin meal

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] (1) Acetone degreasing

[0058] Mix lupine powder and acetone at a ratio of 1:10 (W / V), stir and degrease in a 4°C refrigerator with the help of a magnetic stirrer for 2h, and then centrifuge at 9000r / min at 4°C for 15min with a refrigerated centrifuge to obtain a precipitate; The precipitation was degreased repeatedly 4 times, and after completion, the precipitation was placed in a fume hood to air-dry the acetone overnight, and the degreased lupine powder was obtained after drying;

[0059] (2) Crude extraction by salting out method

[0060] Weigh the degreased lupine flour, add 5mL 50mmol / L Tris-HCL buffer solution containing 1.0mol / L NaCl to extract protein according to 1g of lupine flour, stir well and place it in a refrigerator at 4°C for overnight magnetic stirring. Centrifuge at 11000r / min in a centrifuge for 20min at 4°C, and the crude extract of lupine allergens is the supernatant obtained by centrifugation;

[0061] (3) Fractional precipitation of ammonium ...

Embodiment 2

[0085] A method for extracting β-conglutin from Australian sweet lupine flour, comprising the steps of:

[0086] (1) Acetone degreasing

[0087] Mix lupine powder and acetone at 1:10 (W / V), stir and degrease in a 3°C refrigerator with the help of a magnetic stirrer for 1h, and then centrifuge at 9000r / min at 3°C ​​for 10min with a refrigerated centrifuge to obtain a precipitate; The precipitation was repeatedly degreased 3 times, and after completion, the precipitation was placed in a fume hood to air-dry the acetone overnight, and the degreased lupine powder was obtained after drying;

[0088] (2) Crude extraction by salting out method

[0089] Weigh the degreased lupine flour, add 5mL 50mmol / L Tris-HCL buffer solution containing 1.0mol / L NaCl to extract protein according to 1g of lupine flour, stir evenly, and place it in a refrigerator at 3°C ​​for overnight magnetic stirring. Centrifuge in a centrifuge at 12000r / min for 15min at 3°C, and the crude extract of lupine aller...

Embodiment 3

[0099] A method for extracting β-conglutin from Australian sweet lupine flour, comprising the steps of:

[0100] (1) Acetone degreasing

[0101] Mix lupine powder and acetone at a ratio of 1:10 (W / V), stir and degrease in a 6°C refrigerator with the help of a magnetic stirrer for 3h, and then centrifuge at 9000r / min at 6°C for 20min with a refrigerated centrifuge to obtain a precipitate; The precipitation was repeatedly degreased 5 times, and after completion, the precipitation was placed in a fume hood to air-dry the acetone overnight, and the degreased lupine powder was obtained after drying;

[0102] (2) Crude extraction by salting out method

[0103] Weigh the degreased lupine flour, add 5mL 50mmol / L Tris-HCL buffer solution containing 1.0mol / L NaCl to extract protein according to 1g of lupine flour, stir evenly and place it in a refrigerator at 6°C for overnight magnetic stirring. Centrifuge at 10,000r / min in a centrifuge for 30min at 6°C, and the crude extract of lupin...

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Abstract

The invention relates to a method for extracting sensitized conglutin-beta-conglutin with the highest content and strong sensitization from Australian sweet lupin meal. The lupin meal serves as a raw material and is treated by reagents such as acetone, Tris-HCl and ammonium sulfate, separation of target proteins is promoted by centrifugation, anion exchange chromatography and gel exclusion chromatography, target objects with higher purity are acquired by separation and purification, and the separated proteins are represented by SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) gel electrophoresis. Based on the background, a foundation is laid for subsequent further exploration of protein characters, production of corresponding specific antibodies and research of human toxicity. By the method, target allergens can be selectively separated, high-purity protein components are acquired, and the concentration of lupin can be accurately determined. Besides, toxicity characteristics of the allergens can be illustrated by observing reaction of the extracted allergens and immune globulin E in corresponding serum, and key information is provided for analysis experiments for further detecting lupin allergens in food.

Description

[0001] Technical field: [0002] The invention relates to the technical field of biological materials, in particular to a lupine conglutin-β-conglutin extraction method. [0003] Background technique: [0004] Lupine (scientific name: Lupinus micranthus Guss.), commonly known as Lupine flower, can be divided into edible lupine and ornamental lupine according to its use. Lupines are widely distributed and there are many kinds. Generally speaking, cultivated lupines are generally divided into 5 species: narrow-leaf lupine, also known as Australian sweet lupine, white lupine, aliased as Egyptian lupine, yellow lupine, Sandy plains lupine and South American lupine. In recent years, lupine has attracted much attention as a high-quality food material with high nutritional value. It contains 5.5%-10% oil, 11%-14% dietary fiber, 32%-36% crude protein (the protein) as well as a small amount of starch and a high amount of polygalactose. α-conglutin, β-conglutin, γ-conglutin and δ-cong...

Claims

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Application Information

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IPC IPC(8): C07K14/415C07K1/36C07K1/30C07K1/18C07K1/16C07K1/14G01N27/447
CPCC07K14/415G01N27/447G01N27/44747
Inventor 陆旸张咏王黔王硕
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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