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Separation and purification method of mung bean trypsin inhibitor and its application in the production of surimi products

A trypsin inhibition, separation and purification technology, which is applied to the separation and purification of mung bean trypsin inhibitor and the application field in the production of surimi products, can solve the problems of cumbersome separation and purification process, complicated operation, long time consumption, etc. Sample time, simplified purification steps, simple process effect

Active Publication Date: 2011-12-28
JIMEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the separation and purification of mung bean trypsin inhibitor mainly adopts methods such as sulfuric acid extraction, ammonium sulfate fractional precipitation, membrane ultrafiltration, ion exchange column chromatography, affinity chromatography or high performance liquid chromatography. The separation and purification process is cumbersome and the operation is complicated. Long time-consuming, low yield, high cost

Method used

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  • Separation and purification method of mung bean trypsin inhibitor and its application in the production of surimi products
  • Separation and purification method of mung bean trypsin inhibitor and its application in the production of surimi products
  • Separation and purification method of mung bean trypsin inhibitor and its application in the production of surimi products

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Embodiment 1

[0028] A kind of separation and purification method of mung bean trypsin inhibitor (Mung bean trypsin inhibitor, MBTI) of the present embodiment comprises the steps:

[0029] 1. Extraction: Grind 400 g of mung beans into powder, add 1600 ml of sulfuric acid with a concentration of 0.1 mol / L, and stir for 24 h. Use a Kinematica (Switzerland) tissue masher to mash for 30 min, centrifuge at 12,000 g for 20 min, and collect about 700 mL of the supernatant, which is the crude MBTI extract.

[0030] 2. Neutralization: Add 1 mol / L NaOH to 700 mL crude extract until the pH of the solution is neutral.

[0031] 3. Heat treatment: The neutralized MBTI crude extract was heated in a 60°C water bath for 90 min, cooled immediately after heating, centrifuged at 12000 g for 10 min, and the supernatant was collected.

[0032] 4. Membrane treatment: the above-mentioned MBTI supernatant obtained through heat treatment is ultrafiltered with an ultrafiltration membrane with a molecular weight cut-...

Embodiment 2

[0038] A kind of separation and purification method of mung bean trypsin inhibitor (Mung bean trypsin inhibitor, MBTI) of the present embodiment comprises the steps:

[0039] 1. Extraction: take 100g mung bean powder, add 500 mL 0.1 mol / L H 2 SO 4 Stir and stand for 20 hours, fully mash the tissue, centrifuge at 10,000 g for 20 min, and collect about 170 mL of the supernatant, which is the crude MBTI extract.

[0040] 2. Neutralization: adjust the pH of the crude extract with 2 mol / L NaOH to stabilize the pH of the solution at around 7.

[0041] 3. Heat treatment: heat the neutralized MBTI crude extract in a water bath at 80°C for 30 min, cool immediately after heating, centrifuge at 10000g for 15 min, and collect the supernatant.

[0042] 4. Membrane treatment: The MBTI supernatant obtained by the above heat treatment is ultrafiltered with an ultrafiltration membrane with a molecular weight cut-off of 30 kDa to retain large molecular weight proteins, while the target protei...

Embodiment 3

[0049] A kind of separation and purification method of mung bean trypsin inhibitor (Mung bean trypsin inhibitor, MBTI) of the present embodiment comprises the steps:

[0050] 1. Extraction: Grind 500 g of dried mung beans into powder, add 3,000 ml of sulfuric acid with a concentration of 0.075 mol / L, stir for 36 h, mash the tissue for 30 min, centrifuge at 12,000 g for 20 min, and collect the supernatant About 1250 mL is the crude extract of MBTI.

[0051] 2. Neutralization: Add 1.2 mol / L NaOH to 1250 mL crude extract until the pH of the solution is neutral.

[0052] 3. Heat treatment: Heat the neutralized MBTI crude extract in a water bath at 80°C for 30 min, cool immediately after heating, centrifuge at 12,000 g for 15 min, and collect the supernatant.

[0053] 4. Membrane treatment: the above-mentioned MBTI supernatant obtained through heat treatment is ultrafiltered with an ultrafiltration membrane with a molecular weight cut-off of 30 kDa, and the external liquid is coll...

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Abstract

The invention discloses a separation and purification method of an MBTI. The method comprises the following steps: 1, extraction; 2, neutralization; 3, heat treatment; 4, membrane treatment; and 5, adsorptive separation, and desorption. So the high purity MBTI is obtained. The invention also discloses an application of the MBTI, and the application is that the MBTI, which is added to surimi products treating seawater fish or freshwater fish as a main raw material and is uniformly mixed, is used for enhancing the elasticity of the surimi products. The separation and purification method of the invention has the advantages of simple technology and good separation effect, and the MBTI has a great improvement effect on the elasticity enhancement of the surimi products.

Description

technical field [0001] The invention relates to a separation and purification method of mung bean trypsin inhibitor and its application in the production of surimi products. Background technique [0002] Mung bean trypsin inhibitor (Mung Bean Trypsin Inhibitor, MBTI) is obtained from mung bean ( Phaseolus radiatus L. ) isolated from mature seeds belong to the Bowman-Birk type of serine protease inhibitors. Chinese scholar Qi Zhengwu et al. have deeply and systematically studied the physical, chemical and biochemical properties of MBTI, and determined its primary structure. MBTI contains a total of 72 amino acid residues, a molecular weight of 8,883 Da, and 7 pairs of disulfide bonds. It has high stability to heat, acid, alkali, and enzyme digestion. It has two structural domains with the same inhibitory activity, and two inhibitory active sites (Lys20-Ser21, Arg47-Ser48) both act on trypsin. The former consists of a long chain of 26 residues and a short chain of 9 residue...

Claims

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Application Information

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IPC IPC(8): C07K14/81C07K1/36C07K1/30C07K1/18A23L1/326A23L17/10
Inventor 曹敏杰
Owner JIMEI UNIV
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