Method for separating and purifying plant ice structural protein

An ice structure protein, separation and purification technology, applied in the field of food additive separation, can solve the problems of high cost, unable to maintain a reasonable price, loss of antifreeze activity, long separation cycle, etc. The effect of industrial production

Inactive Publication Date: 2009-11-18
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have the disadvantages of long separation cycle, cumbersome process and low yield, and in large-scale production, the product cannot be maintained at a reasonable price due to high cost.
Chinese patents (CN1117497A

Method used

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  • Method for separating and purifying plant ice structural protein
  • Method for separating and purifying plant ice structural protein
  • Method for separating and purifying plant ice structural protein

Examples

Experimental program
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Effect test

Embodiment 1

[0021] Weigh 100 g of pulverized winter wheat, add 1000 mL of phosphate buffer solution (100 mM, pH 7.3) containing 0.5% Genapol X-080 (fatty alcohol polyoxyethylene ethers, purchased from Sigma-Aldrich Company), and soak for 1 hour Filtrate to obtain the leach solution of solid filter residue and ice-structural protein; continue to add Genapol X-080 to 5% to the ice-structural protein leach solution, then heat to 50°C for phase-separation extraction, centrifuge at 5000rpm for 10min after 2h, and separate Take out the water phase; take the water phase solution and place it in a dialysis bag with a molecular weight cut-off of 6000 Da, and fully dialyze at 4°C. After dialysis, use polyethylene glycol 10,000 to cover and concentrate the dialysis bag, concentrate to 1 / 20 of the original volume, and then freeze After drying, 2.8 g of the isolated and purified ice structural protein sample was obtained. Accurately weigh 0.5 g of the above ice-structured protein sample, dissolve it i...

Embodiment 2

[0025] Weigh 100 g of crushed winter wheat, add 1000 mL of phosphate buffer solution (100 mM, pH 7.3) containing 1% Triton X-114 (alkylphenol polyoxyethylene ethers, purchased from Shanghai Jingchun Reagent Co., Ltd.), and extract Filter after 1 hour to obtain the solid filter residue and the leach solution of ice-structural protein, continue to supplement Triton X-114 to 3% in the ice-structural protein leach solution, then heat to 35°C for phase-separation extraction, centrifuge at 5000rpm after 2h After 10 minutes, the water phase was separated, and the water phase solution was placed in a dialysis bag with a molecular weight cut-off of 6000 Da, and fully dialyzed at 4°C. After dialysis, the dialysis bag was covered and concentrated with polyethylene glycol 10,000, and concentrated to 1 / 20 of the original volume. Freeze-drying was then carried out to obtain 3.5 g of the isolated and purified ice-structural protein sample. Accurately weigh 0.5 g of the above ice-structured p...

Embodiment 3

[0029]Take pulverized winter privet leaves 100g, add 1000mL containing 0.5% Genapol X-080 (fatty alcohol polyoxyethylene ethers, purchased from Sigma-Aldrich company) phosphate buffer solution (100mM, pH 7.3), leaching Filtrate after 2 hours to obtain the solid filter residue and the leach solution of ice-structural protein; continue to supplement Genapol X-080 to 5% in the ice-structural protein leach solution, then heat to 50°C for phase-separation extraction, and centrifuge at 5000rpm after 2h After 10 minutes, the aqueous phase was separated; the aqueous phase solution was placed in a dialysis bag with a molecular weight cut-off of 6000 Da, and fully dialyzed at 4°C. After dialysis, the dialysis bag was covered and concentrated with polyethylene glycol 10,000, and concentrated to 1 / 20 of the original volume. Freeze-drying was then carried out to obtain 1.8 g of the isolated and purified ice-structural protein sample. Accurately weigh 0.5 g of the above ice-structured prote...

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Abstract

The invention relates to a method for separating and purifying plant ice structural protein, which belongs to the field of food additive separation. The method uses cold-stress plants as raw materials to separate and purify the ice structural protein by leaching and adopting a cloud point extraction method, and comprises the following steps: firstly, adopting buffer solution containing a non-ionic surfactant to leach out the ice structural protein from the plant raw materials; secondly, performing cloud point extraction by adopting a micellar solution formed by the non-ionic surfactant, wherein the mixture is divided into a water phase and a micellar phase after being heated to the cloud point temperature, hydrophobic impurities are enriched at the micellar phase, while the hydrophilic ice structural protein is kept at the water phase; and finally, performing dialysis concentration and freeze drying to obtain a white ice structural protein mixture. A differential scanning calorimetry is adopted to perform thermal hysteresis activity detection on a separated and purified sample to prove that the product has high thermal hysteresis activity. The method has the advantages of simple separation process, low production cost, no volatile organic solvent in the process of extraction and high purification efficiency, and is advantageous for producing the ice structural protein industrially and making full use of plant resources.

Description

technical field [0001] The invention discloses a method for separating and purifying plant ice structural protein, relates to a method for separating and purifying ice structural protein from cold stress plants, and belongs to the field of separation of food additives. Background technique [0002] Antifreeze or ice-structuring proteins are a class of proteins that inhibit the growth and recrystallization of ice crystals. It is distributed in cold zone plants, marine fish, insects and microorganisms, and at extremely low concentrations, it can improve the ability of organisms to resist cold and avoid damage to the organism caused by the growth of ice crystals. This undoubtedly has positive significance for reducing the ice crystal damage of frozen food, maintaining the texture and performance of frozen food, and prolonging the storage period of products. It non-colligatively lowers the freezing point of aqueous solutions below the melting point (the melting point is always ...

Claims

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Application Information

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IPC IPC(8): A23J3/22A23J3/14A23L3/3526A23L3/375
Inventor 徐化能黄卫宁陈海英
Owner JIANGNAN UNIV
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