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Antifreeze protein and method for extracting antifreeze protein from gluten

A technology of antifreeze protein and gluten, which is applied in the field of antifreeze protein and extracting antifreeze protein from gluten, can solve the problems that raw materials are not easy to obtain and are not suitable for large-scale production, so as to improve the utilization value and protein The effect of rich content and simple operation process

Inactive Publication Date: 2016-10-12
TIANJIN UNIV OF COMMERCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The existing plant-based antifreeze protein extraction is mainly concentrated in alpine snow lotus, ryegrass, Inner Mongolia holly and other plant raw materials in alpine and alpine regions, but these raw materials are not easy to obtain, and the extraction method is mainly "ice finger method". suitable for mass production

Method used

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  • Antifreeze protein and method for extracting antifreeze protein from gluten
  • Antifreeze protein and method for extracting antifreeze protein from gluten
  • Antifreeze protein and method for extracting antifreeze protein from gluten

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] (1) Extraction and preparation of crude antifreeze protein

[0031] Weigh 10g of gluten powder and dissolve it in 150ml of 0.09mol / L NaOH solution with a pH value of 11, keep the extraction temperature at 55°C in a water bath and stir at a constant temperature for 2h, then extract it at 7500r / min at 4°C Centrifuge for 15 minutes, take the supernatant, dialyze overnight in a 7Ku dialysis bag, freeze-dry for 24 hours, and obtain 1.837 g of crude antifreeze protein, and the yield of crude antifreeze protein is 18.37%.

[0032] (2) Purification of crude antifreeze protein

[0033] The crude antifreeze protein extracted and separated was redissolved in 10mmol / L Tris-Hcl buffer with a pH value of 8.5 to prepare a 20mg / ml crude protein solution, which was chromatographically separated by SepHadex G-100 gel chromatography. Gel chromatographic separation chart such as figure 1 As shown, the column size used in the chromatography is φ1.0cm×10cm, pre-equilibrated with 10mmol / L T...

Embodiment 2

[0047] (1) Extraction and preparation of crude antifreeze protein

[0048] Weigh 5g of gluten powder and dissolve it in 100ml of 0.07mol / L Tris-Hcl buffer solution with a pH value of 8.2, keep the extraction temperature at 30°C in a water bath and stir at a constant temperature for 2.5h, and then at 8000r / min, Centrifuge at 4°C for 20 minutes, take the supernatant, dialyze overnight in a 7Ku dialysis bag, freeze-dry for 24 hours, and obtain 0.972 g of crude antifreeze protein, and the yield of crude antifreeze protein is 9.72%.

[0049] (2) Purification of crude antifreeze protein

[0050] The crude antifreeze protein extracted and separated was redissolved in 10mmol / L, pH=8.0 Tris-Hcl buffer to prepare a 20mg / ml crude protein solution, which was separated by SepHadex G-100 gel chromatography. Chromatographic separation and figure 1 Basically the same, the column size used in the chromatography is φ1.0cm×10cm, pre-balanced with 10mmol / L, pH=8.0 Tris-Hcl buffer, 150μL of samp...

Embodiment 3

[0056] (1) Extraction and preparation of crude antifreeze protein

[0057] Weigh 15g of gluten powder and dissolve it in 150ml of 0.01mol / L PBS buffer solution with a pH value of 8.9, keep the extraction temperature at 25°C in a water bath and stir at a constant temperature for 4h, then in a water bath at 8500r / min, 4°C Centrifuge for 10 min under the conditions, take the supernatant, dialyze overnight in a 7Ku dialysis bag, and freeze-dry for 24 hours to obtain 0.863 g of crude antifreeze protein, and the yield of crude antifreeze protein is 8.63%.

[0058] (2) Purification of crude antifreeze protein

[0059] The crude antifreeze protein extracted and separated was redissolved in 10mmol / L, pH=8.2 Tris-Hcl buffer to prepare a 20mg / ml crude protein solution, which was separated by SepHadex G-100 gel chromatography. Chromatographic separation and figure 1 Basically the same, the column size used in the chromatography is φ1.0cm×10cm, pre-balanced with 10mmol / L, pH=8.2 Tris-Hcl...

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Abstract

The invention discloses an antifreeze protein and a method for extracting the antifreeze protein from vital gluten, and provides an antifreeze protein and an extraction method thereof which is simple to operate, and easy to industrially produce on large scale. The molecular weight of the antifreeze protein is 43.14KDa, and the thermal hysteresis activity is 0.08 DEG C by measurement of a differential scanning calorimeter. The antifreeze protein is prepared by the following steps: taking vital gluten as a raw material; centrifuging, carrying out dialysis desalination, freezing and drying to obtain an antifreeze protein coarse product after extracting by an alkaline solution, wherein the concentration of the alkaline solution is 0.01-0.09 mol / L, and the pH value is 8-11. A by-product vital gluten produced by wheat starch is taken as the raw material, so that the vital gluten is wide in source, low in price and available, and abundant in protein content, and the cost of the antifreeze protein can be reduced. Poisonous and harmful substances are not added in the preparation process, meanwhile, the vital gluten is an edible plant raw material, and the obtained antifreeze protein is a natural product. Therefore, the antifreeze protein can be applied to a food industry in a form of an additive.

Description

technical field [0001] The invention relates to the technical field of food engineering, in particular to an antifreeze protein and a method for extracting the antifreeze protein from gluten. Background technique [0002] Low temperature stress is a serious natural disaster. It is fatal to most organisms. It leads to dehydration and crystallization of the intracellular environment, causing physical damage or death of cells. The annual loss due to low temperature stress is as high as thousands One hundred million U.S. dollars. [0003] On the other hand, in the process of coagulation, freezing, transportation, storage and sales of current quick-frozen foods such as ice cream, the more cooling capacity is consumed, the smaller the ice crystals of the product can be guaranteed. Fluctuating up and down will still cause the ice crystals to grow and result in a rough taste. If there is a substance that can control the fluidity of water molecules from the beginning, and with the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C07K1/14
CPCC07K14/415
Inventor 刘爱国赵源吴子健
Owner TIANJIN UNIV OF COMMERCE
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