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A set of peptide markers for evaluating the quality of live oysters

A technology of markers and live oysters, applied in the field of food science, can solve problems such as evaluating the quality of live oysters

Active Publication Date: 2021-11-26
山东佛坤投资有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no way to evaluate the quality of live oysters by detecting endogenous differential peptides

Method used

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  • A set of peptide markers for evaluating the quality of live oysters
  • A set of peptide markers for evaluating the quality of live oysters
  • A set of peptide markers for evaluating the quality of live oysters

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Screening of polypeptide markers:

[0032] (1) Sample information:

[0033] There are 30 fresh oysters just caught and 30 oysters whose quality has been declining over time after harvesting.

[0034] (2) Sample pretreatment steps:

[0035] (1) Shell the oysters, take the edible parts, freeze them in liquid nitrogen, and homogenize them. Weigh 20 g of the homogenized oyster samples into a powder state, heat at 95°C for 10 minutes, and add 20ml of ammonium bicarbonate solution at 4°C, 15000 r / min. Centrifuge for 15 min, collect the supernatant, and obtain the crude peptide extract;

[0036] (2) Add 100 µL of 1 mol / L Dithiothreotol (DTT) to the above protein solution, shake in a water bath at 60°C for 30 min, and then cool to room temperature;

[0037] (3) Take 1000 µL of 1 mol / L freshly prepared iodoacetamide (IAA) solution and react at room temperature for 1 h in the dark;

[0038] (4) Desalting the above solution using a solid-phase extraction cartridge; ...

Embodiment 2

[0054] Example 2 Synthesis of identified differential peptides and establishment of a mass spectrometry multiple reaction monitoring method for verification:

[0055] (1) The above-mentioned peptides are synthesized by a third-party company and are ready for use; and the m / z and product ions of the polypeptide markers are shown in the following table:

[0056] SEQ ID NO.1: SSSTGEVGTYSGTTN;

[0057] SEQ ID NO.2: TARNEANVNI;

[0058] SEQ ID NO.3: VGIIKGSSSEEA;

[0059] SEQ ID NO. 4: TARNEANVNIY.

[0060] Table 1. m / z and product ions of peptide markers SEQ ID NO.1-4

[0061]

[0062] Reconstitute the peptide, and before further analysis on the machine, transfer the reaction solution to a centrifuge tube equipped with a 10 kDa ultrafiltration membrane, and centrifuge at 8000 r / min for 20 minutes at room temperature to collect the peptide filtrate in the lower layer and wait for the test on the machine ;

[0063] (2) On-board testing:

[0064] Using AB SCIEX 5500 triple q...

Embodiment

[0071] Example 3 Expression of the above-mentioned peptides in oysters of different qualities:

[0072] (1) Sample information:

[0073] The oysters were collected from a farm in Rongcheng City, Shandong Province, and transported to the laboratory on the same day under ice storage conditions. After washing with running water, they were temporarily kept and purified, and the experiment of keeping alive at 4°C was carried out. Samples were taken on the 1st, 3rd, 5th, 7th, and 9th days of the shelf life, and 30 oysters were taken each time. After oysters leave the water body, the degree of freshness shows a downward trend. Therefore, sampling and analysis of oysters collected at different times can have a greater correlation with oyster quality evaluation.

[0074] (2) Sample pretreatment steps:

[0075] (1) Shell the oysters, take the edible parts, freeze them in liquid nitrogen, and homogenize them. Weigh 20 g of the homogenized oyster samples into a powder state, heat at 95°...

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Abstract

The invention discloses a group of polypeptide markers for evaluating the quality of live oysters. The sequence of the polypeptide is: SEQ ID NO.1: SSSTGEVGTYSGTTN; SEQ ID NO.2: TARNEANVNI; SEQ ID NO.3: VGIIKGSSSEEA; SEQ ID NO.4: TARNEANVNIY. For the first time, the present invention uses peptidomics technology to comprehensively study the endogenous peptides of oysters of different qualities, and obtains a set of polypeptide markers that can be used to evaluate the quality of live oysters. The multiple reaction monitoring analysis method established by stage mass spectrometry verifies the correctness of the marker peptide sequence and the applicability for routine liquid mass analysis. It can supplement the mechanism of quality change of live oysters and improve the evaluation system of quality change in the process of keeping alive.

Description

technical field [0001] The invention belongs to the field of food science, and in particular relates to a group of polypeptide markers for evaluating the quality change of live oysters. Background technique [0002] Oyster is an important marine aquatic resource. It is rich in nutrition and fresh and tender, and is deeply loved by consumers. With the changes in the product structure of shellfish seafood in the world, fresh and live oysters are the most popular in domestic and foreign markets, especially with the improvement of people's consumption level, the demand for fresh and live shellfish in some inland areas is also increasing high. Therefore, we need to optimize the fresh supply chain of live oysters. The premise of improving the live oyster supply chain is to establish an accurate quality evaluation method for live oysters. However, due to the complexity of changes in living organisms, there is still a lack of reliable quality evaluation methods for live oysters, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06C07K7/08G01N30/02
CPCC07K7/06C07K7/08G01N30/02G01N2410/00
Inventor 李兆杰陈李品张鸿伟张晓梅胡玲萍蒋冰雪薛勇薛长湖
Owner 山东佛坤投资有限公司
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