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Preparation method for umami peptide affinity column

A peptide-affinity and umami-taste technology, which is applied in the field of preparation of umami-taste peptide affinity columns, can solve the problems of complex separation steps, less pure products, and difficulty in maintaining activity, and achieve simple and easy operation, high purification efficiency, and column The effect of large capacity

Active Publication Date: 2016-03-09
ZHEJIANG ACAD OF MEDICAL SCI
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  • Claims
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Problems solved by technology

[0004] At present, the separation and purification methods of umami peptides generally adopt precipitation, electrophoresis, dialysis, chromatography and centrifugation, etc. The main defects are: (1) It takes a long time and the efficiency is not high (2) The purity of the separated umami peptides is not high (3) The separation steps are complicated, the resulting pure product is less, and the activity is difficult to maintain
[0008] An affinity column that specifically adsorbs umami-taste peptides using umami-taste receptor proteins has not been reported yet

Method used

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  • Preparation method for umami peptide affinity column
  • Preparation method for umami peptide affinity column
  • Preparation method for umami peptide affinity column

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Embodiment 1

[0028] Embodiment 1: The preparation method of the umami peptide affinity column of this embodiment comprises the following steps:

[0029] (1) Purification of receptor proteins T1R1 and T1R3

[0030] The T1R1 / T1R3 gene is synthesized by gene recombination, the gene sequence of T1R1 is shown in SEQ ID NO.1, and the gene sequence of T1R3 is shown in SEQ ID NO.2;

[0031] Subclone the T1R1 / T1R3 gene obtained in step (1) into the pcDNA3.1(+)[EF550208.1] vector, express it in mammalian cells, purify His, and obtain the receptor protein T1R1 / T1R3; the purified Receptor protein T1R1 / T1R3 as figure 1 and 2shown. It shows that the column material of umami peptide affinity column can be prepared by gene recombination method, and it can specifically adsorb umami peptide, so as to ensure the purification effect of the affinity column.

[0032] (2) Affinity column preparation

[0033] 2.1 Washing

[0034] Suspend 12 g of agarose dry gel activated by cyanogen bromide (CNBr) in 120 mL...

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Abstract

The present invention discloses a preparation method for a umami peptide affinity column. The method comprises the following steps: synthesizing receptor proteins T1R1 and T1R3; taking protein T1R1 / T1R3 agarose gel as a solid phase support; coupling the protein T1R1 / T1R3 on the support with umami peptide; and chemically bonding the coupled umami peptide with a stationary phase. The preparation method for the affinity column is simple and reusable; purification efficiency is improved, so that the sensitivity of the method is improved; theoretically the method is applicable to various sample matrices; and due to the protein receptors T1R1 / T1R3, the method has the good separation effect on the umami peptide, high coupled specificity, a large adsorption amount and can be used repeatedly.

Description

technical field [0001] The invention relates to a preparation method of an umami peptide affinity column. Background technique [0002] In recent years, the theory and application of the taste function of peptides has attracted widespread attention from scholars from all over the world. Umami peptides are the dominant ingredients in the taste of umami foods. Using traditional column chromatography, scholars have isolated several protein peptides with umami or umami-enhancing effects from meat, cheese, seafood and crops. For example, Noguchi et al. Polypeptides Glu-Glu, Ser-Glu-Glu, Glu-Ser, Thr-Glu, Glu-Asp, Asp-Glu-Ser and flavor enhancing peptides Gly-Leu, Pro-Glu, Val-Glu, B-Ala-His . Yamasaki et al. used papain to hydrolyze beef, separated and purified through gel filtration and ion exchange chromatography, and obtained the octapeptide Lys-Gly-Asp-Glu-Glu-Ser-Leu-Ala from the hydrolyzate, which itself does not have Meat taste, but it has a better synergistic effect w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D15/38B01D15/20C07K1/22
Inventor 党亚丽高新昌包国良李青青吴庆其
Owner ZHEJIANG ACAD OF MEDICAL SCI
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