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Method for extracting ice structuring protein from clovers

A technology of ice-structural protein and alfalfa, which is applied to the preparation method of peptides, chemical instruments and methods, plant peptides, etc., can solve the problems that ice-structural proteins cannot be widely used, the extraction raw materials of ice-structural proteins are limited, and the extraction process is complicated, etc., to achieve The effect of less impurities, low cost and simple extraction process

Inactive Publication Date: 2016-11-16
HARBIN UNIV OF COMMERCE
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Problems solved by technology

[0006] In order to solve the problems in the prior art that the ice structural protein extraction raw materials are limited, the extraction process is complicated, the cost is high, and the extracted ice structural protein cannot be widely used in frozen food and medicine, a method for extracting ice structural protein from alfalfa is proposed. Methods for Structural Proteins

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  • Method for extracting ice structuring protein from clovers
  • Method for extracting ice structuring protein from clovers

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specific Embodiment approach 1

[0026] Specific embodiment one: the method for extracting ice structural protein from alfalfa in the present embodiment is carried out according to the following steps:

[0027] 1. Alfalfa hay treatment

[0028] Get the alfalfa hay, remove impurities and dust in the alfalfa hay, discard the grass stems, keep the alfalfa hay leaves, crush them and pass through an 80-mesh sieve for later use;

[0029] 2. Preparation of ice puck

[0030] Place a glass ball with a diameter of 1 cm in each ice hole of the silicone ice tray, pour distilled water into the ice hole, and then freeze in a -18°C refrigerator to obtain an ice ball with a diameter of 2 cm;

[0031] 3. Supernatant preparation

[0032] The ratio of solid to liquid is 1g: (5-30mL) mix the alfalfa powder prepared in step 1 with phosphate buffer solution to obtain a mixed solution, place the obtained mixed solution at room temperature, stir and extract for 0.5-3.0h, and then The extracted mixed solution is placed in a centri...

specific Embodiment approach 2

[0042] Embodiment 2: This embodiment is different from Embodiment 1 in that: the moisture content of the alfalfa hay described in Step 1 is 1.3%. Other steps and parameters are the same as those in the first embodiment.

specific Embodiment approach 3

[0043]Embodiment 3: The difference between this embodiment and Embodiment 1 or 2 is that the silica gel ice tray described in step 2 is a silica gel ice tray with a temperature resistance of -40°C, and the silica gel ice tray is plate-shaped, 24 cm long and 24 cm wide. 7.3cm, 10×2 spherical ice holes are arranged on the board, and the inner diameter of each ice hole is 2cm. Other steps and parameters are the same as those in Embodiment 1 or 2.

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Abstract

The invention relates to a method for extracting ice structuring protein from clovers. The invention aims to solve the problems of the prior art that the extraction raw materials of the ice structuring protein are limited, the extraction process is complex, the cost is relatively high, and the extracted ice structuring protein can not be widely used in frozen food and medical science. The extraction method comprises the following steps: (1) taking dry leaves of the clovers, and grinding and sieving; (2) preparing an ice ball of which the diameter is 2cm; (3) mixing alfalfa meal and a phosphate buffer solution, and centrifuging; (4) adding the ice ball into supernatant liquid, and extracting; and (5) extracting the ice structuring protein again. The method provided by the invention has the advantages that the extraction concentration is high, impurities are few, the cost and time are saved, the molecular weight can be determined by directly performing SDS-PAGE, cold induction is not needed, the raw materials are convenient and easy to obtain, the cost is low, and the extraction process is simple and can be universally applied to frozen food. The method is suitable for extracting the ice structuring protein from the clovers.

Description

technical field [0001] The invention relates to a method for extracting ice structural protein from alfalfa. Background technique [0002] Ice structuring proteins (ISPs), also known as antifreeze proteins and antifreeze proteins (AFPs), are a class of proteins produced by certain fish, insects, plants, fungi and bacteria to resist external environmental stress. The polypeptide produced by the reaction has the ability to prevent the formation of ice crystals, modify the shape of ice crystals and inhibit recrystallization. It has a certain thermal hysteresis activity, that is, it can lower the freezing point of the solution in a non-colligative form, but has little effect on the melting point. This creates a difference between the freezing point and melting point of the solution. The unique properties of ice structural proteins ensure the survival of organisms under low temperature conditions, and they have the ability to specifically bind to ice. In 2006, the Ministry of H...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C07K1/14
CPCC07K14/415
Inventor 曲敏刘羽佳李凌俐卢曼曼耿浩源
Owner HARBIN UNIV OF COMMERCE
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