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Method for rapidly detecting and quantifying activity of proteinase in dry-cured ham processing process

A protease activity, dry-cured ham technology, applied in the field of analytical chemistry, can solve problems such as undeveloped, achieve the effect of improving efficiency, reducing labor intensity, and accurate experimental results

Inactive Publication Date: 2017-05-24
夏云
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

No method currently developed for rapid detection and quantification of cathepsin activity during dry-cured ham processing

Method used

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  • Method for rapidly detecting and quantifying activity of proteinase in dry-cured ham processing process
  • Method for rapidly detecting and quantifying activity of proteinase in dry-cured ham processing process
  • Method for rapidly detecting and quantifying activity of proteinase in dry-cured ham processing process

Examples

Experimental program
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Embodiment 1

[0041] A method for rapidly detecting and quantifying protease activity during dry-cured ham processing, comprising the steps of:

[0042] Step (1), making gelatin smears: add potassium chromium sulfate dodecahydrate and gelatin to water, heat to 68°C, put a clean sterile three-well glass slide in it after the gelatin is completely dissolved, and keep the temperature After 10 minutes, the three-well glass slide was taken out and dried naturally to obtain a gelatin smear; wherein, the mass ratio of potassium chromium sulfate dodecahydrate, gelatin and water was 0.43:2.4:3.5×10 3 ;

[0043] Step (2), sample collection and dilution: Under sterile conditions, cut the ham into pieces with a particle size of 2-4 cm for later use; then add sterilized 1×PBS with a pH of 7.0 to the ham pieces Buffer solution, after mixing, stir at 16000r / min for 1.5 minutes, then grind at 10000r / min for 1 minute to obtain meat paste; wherein, the mass-volume ratio of ham pieces to 1×PBS buffer is 1: ...

Embodiment 2

[0052] A method for rapidly detecting and quantifying protease activity during dry-cured ham processing, comprising the steps of:

[0053] Step (1), making gelatin smears: add potassium chromium sulfate dodecahydrate and gelatin to water, heat to 72°C, put a clean sterile three-well glass slide in it after the gelatin is completely dissolved, and keep the temperature After 15 minutes, the three-well glass slide was taken out and dried naturally to obtain a gelatin smear; wherein, the mass ratio of potassium chromium sulfate dodecahydrate, gelatin and water was 0.45: 2.6: 3.5×10 3 ;

[0054] Step (2), sample collection and dilution: Under sterile conditions, cut the ham into pieces with a particle size of 2-4 cm for later use; then add sterilized 1×PBS with a pH of 7.0 to the ham pieces Buffer, after mixing, stir at a speed of 18000r / min for 2.5 minutes, and then grind at a speed of 13000r / min for 3 minutes to obtain a meat paste; wherein, the mass-volume ratio of ham pieces t...

Embodiment 3

[0064] A method for rapidly detecting and quantifying protease activity during dry-cured ham processing, comprising the steps of:

[0065] Step (1), making gelatin smears: add potassium chromium sulfate dodecahydrate and gelatin to water, heat to 70°C, put a clean sterile three-well glass slide in it after the gelatin is completely dissolved, and keep the temperature , after 13 minutes, take out the three-well glass slide, and dry naturally to obtain a gelatin smear; wherein, the mass ratio of potassium chromium sulfate dodecahydrate, gelatin and water is 0.44:2.5:3.5×10 3 ;

[0066] Step (2), sample collection and dilution: Under sterile conditions, cut the ham into pieces with a particle size of 2-4 cm for later use; then add sterilized 1×PBS with a pH of 7.0 to the ham pieces Buffer solution, after mixing, stir at 17000r / min for 2 minutes, then grind at 12000r / min for 2 minutes to obtain meat puree; wherein, the mass-volume ratio of ham pieces to 1×PBS buffer is 1: 10;

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Abstract

The invention relates to a method for rapidly detecting and quantifying the activity of proteinase in a dry-cured ham processing process. The method comprises the following six steps: manufacturing a gelatin smear, collecting and diluting a sample, eluting the proteinase, collecting the proteinase, determining the activity of cathepsin, and observing through a microscope and determining the activity of the cathepsin. According to the method provided by the invention, the proteinase is calibrated and quantified by adopting a green fluorescent proteinase dyeing method, the activity of the proteinase in the dry-cured ham processing process can be rapidly monitored and quantified, and a complicated protein degradation process in the ham processing process is monitored; salinity, water activity, temperature and humidity conditions in the processing process can be adjusted in time according to the activity of the proteinase, so as to realize quality regulation and control of dry-cured hams; a production and processing process of the dry-cured hams is improved and the curing of the dry-cured hams is accelerated, so that the method provided by the invention has important practical significance on facilitation of industrial production of the dry-cured hams in China.

Description

technical field [0001] The invention belongs to the technical field of analytical chemistry, and in particular relates to a method for rapidly detecting and quantifying protease activity during dry-cured ham processing. Background technique [0002] Traditional dry-cured ham is one of the most important meat products. After a long processing process, a large number of physical, chemical and biochemical reactions have occurred inside, making it a product with a unique flavor. Protein is the main chemical component of traditional dry-cured ham, and its changes directly affect the quality of dry-cured ham. An important indicator of the quality of dry-cured ham is flavor. During the production and processing of dry-cured ham, slow lipid and protein degradation occurred, which led to the accumulation of free fatty acids and free amino acids in ham, respectively, and the above two types of compounds further reacted to form volatile and water-soluble flavor substances. Significan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/37
CPCC12Q1/37
Inventor 夏云孔云虹念馨张旭祥郑旴
Owner 夏云
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