Proteinase K freeze drying technology

A protease and freeze-drying technology, which is applied in the freeze-drying process of proteinase K, can solve the problems of long time spent and affecting the production of proteinase K freeze-dried products, and achieve the effect of improving efficiency, ensuring quality, preservation and stability

Inactive Publication Date: 2017-09-19
GUANGZHOU HEAS BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing proteinase K freeze-drying process is still insufficient, an

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The freeze-drying process operating method of embodiment 1 proteinase K:

[0024] A. Pretreatment: Dissolve 50g of proteinase K in 1L of purified water, and divide it into 5ml vials at 1ml / bottle as the sample to be freeze-dried;

[0025] B. Turn on the freeze-drying machine, adjust the temperature in the freeze-drying chamber to below the first temperature of -50°C, put the pretreated proteinase K into the freeze-drying machine for rapid freezing; adjust the temperature in the freeze-drying chamber to -10°C, and keep freeze-drying The room temperature is -10°C, and the product is kept at a pressure of 60Pa for 2 hours, then cooled to below -50°C, and kept for 2 hours to pre-freeze proteinase K;

[0026] C. Adjust the temperature in the freeze-drying chamber to the second temperature -40°C, and keep the sample at -40°C for 2 hours under a pressure of 10 Pa;

[0027] D. Reduce the temperature of the condenser to -40°C, adjust the temperature in the freeze-drying chamber...

Embodiment 2

[0032] Embodiment 2: within the allowable range, change the freeze-drying process of proteinase K under the parameters such as freeze-drying temperature and time Operation method:

[0033] A. Pretreatment: Dissolve 50g of proteinase K in 1L of purified water, and divide it into 5ml vials at 1ml / bottle as the sample to be freeze-dried;

[0034] B. Turn on the freeze dryer, then put the proteinase K solution sample into the freeze dryer, adjust the temperature to the first temperature below -50°C and keep the temperature below -50°C, freeze the proteinase K, and then raise the temperature to - Keep at 15°C for 1 hour, then lower the temperature to below -50°C, and keep for 1 hour to prefreeze proteinase K;

[0035] C. Adjust the temperature in the freeze-drying chamber to the second temperature of -45°C, and keep the sample at -45°C for 1 hour under a pressure of 20 Pa;

[0036] D. Reduce the temperature of the condenser to -40°C, adjust the temperature in the freeze-drying cha...

Embodiment 3

[0042] Example 3: Application of lyophilized proteinase K in the nucleic acid extraction operation of the lyophilization process of the present invention.

[0043] 1. Virus sample nucleic acid extraction

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PUM

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Abstract

The invention discloses a proteinase K freeze drying technology which comprises the following steps: pretreating proteinase K to be freeze dried, performing pre-freezing treatment on the proteinase K, then heating to a second temperature to quickly drying for the first time and then heating temperature to a third temperature to desorption drying; then reducing pressure, cooling to the second temperature, then haeting to the third temperature and performing special technologies of secondary drying and desorption drying. Compared with the prior art, freeze drying time of the proteinase K freeze drying technology is shortened to 20 to 26 hours; furthermore, a freeze dried product prepared in the proteinase K freeze drying technology has appearance shape meeting a standard, can redissolve quickly and has good stability; proteinase K activity cannot decrease after the proteinase K is stored in normal temperature for many years.

Description

technical field [0001] The invention relates to the field of freeze-drying, in particular to the freeze-drying process of proteinase K. Background technique [0002] Proteinase K is a highly active protease of the subtilisin class used for the general degradation of proteins in biological samples. Purified from Tritirachium album limber. According to the data, the enzyme has two ca2+ binding sites, which are a certain distance away from the active center of the enzyme, and have no direct relationship with the catalytic mechanism. However, if Ca2+ is removed from the enzyme, about 80% of the catalytic activity is lost due to long-range structural changes, but the remaining activity is usually sufficient to degrade proteins that normally contaminate acid preparations. [0003] As a protease, proteinase K can digest and degrade other proteins, but it is also a protein itself. Like most proteins, its activity and stability are also affected by factors such as temperature, pH ...

Claims

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Application Information

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IPC IPC(8): C12N9/50
CPCC12N9/641C12Y304/22038
Inventor 肖湘文陈华云刘淑园叶映玲
Owner GUANGZHOU HEAS BIOTECH CO LTD
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