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Preparation method of high-purity kallidinogenase

A kallikrein and high-purity technology, which is applied in the field of preparation of high-purity kallikrein, can solve the problems of uneven product purity, inability to meet national standards, and low technological level, and achieve unique and novel methods. Safe to use and high level of craftsmanship

Inactive Publication Date: 2016-04-13
CHANGZHOU YAHUAN ENVIRONMENTAL PROTECTION TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem mainly solved by the present invention: In the process of purifying pancreatic kininogenase in the current traditional method, although the pancreatic kininogenase can be purified, there are still low technological levels, and the degree of product purity is not high enough. Balanced, poor quality stability, resulting in the status quo that can not meet the national indicators, provides a tea seed extract mixed with humus sludge, filtered to obtain a filtered liquid, put into tea seed bran, steamed and extruded to make tea seed bran Then collect pig pancreas and saliva, after mixing, program temperature rise, and mix with tea seed bran cake in turn, gradually increase the amount during the mixing process, and use different concentrations of deionized water and sodium chloride to carry out three rinses respectively, Then collect the eluent, adjust the pH value with sodium hydroxide, add ethanol to filter, and finally filter the obtained filtrate with suction, and dry it to obtain the preparation method of high-purity pancreatic kininogenase

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0026] First, take 7kg of rural river silt with a thickness of 7cm, place it in a dryer, and dry it at 65°C. When the water content in the river silt reaches 10%, stop drying, take it out and put it in a container, and place it in a cool place to cool down to Room temperature, and make it layer up and down, then use a scraper to gently scrape off the upper layer of river silt after 4cm to get the lower layer of river channel humus sludge; then take 500g of dry tea seeds without mold, put them in clean water and wash them repeatedly to remove surface residues After the dirt, spread it on a steel sieve covered with gauze, place it in a dryer, and dry it at 100°C for 15 minutes, so that the water content of the tea seeds is 3%, and then put the dried tea seeds into a beaker , add 800mL of ethanol solution with a mass concentration of 75%, mix evenly, place in an ultrasonic oscillator, set the ultrasonic oscillation frequency to 20KHz, ultrasonic power to 100W, ultrasonic oscillati...

example 2

[0030]First, take 7.5kg of rural river silt with a thickness of 9 cm, place it in a dryer, and dry it at 68°C. When the water content in the river silt reaches 13%, stop drying, take it out and put it in a container, and put it in a cool place to cool it by itself to room temperature, and make it layer up and down, and then use a scraper to gently scrape off the upper 7cm of river silt to get the lower layer of river humus sludge; then take 550g of dry tea seeds without mold, put them in clean water and wash them repeatedly to remove the surface After the dirt remains, lay it flat on a steel sieve covered with gauze, place it in a dryer, and dry it for 20 minutes at a temperature of 103°C, so that the water content of the tea seeds is 4%, and then put the dried tea seeds into the In the beaker, add 900mL of ethanol solution with a mass concentration of 75%, mix evenly, place it in an ultrasonic oscillator, set the ultrasonic oscillation frequency to 23KHz, and the ultrasonic po...

example 3

[0033] First take 8kg of rural river silt with a thickness of 10cm, place it in a dryer, and dry it at 70°C. When the water content in the river silt is 15%, stop drying, take it out and put it in a container, and place it in a cool place to cool down to Room temperature, and make it layered up and down, then use a scraper to gently scrape off the upper layer of river silt after 8cm to get the lower layer of river channel humus sludge; then take 600g of dry tea seeds without mold, put them in clean water and wash them repeatedly to remove surface residues After the dirt is removed, lay it flat on a steel sieve covered with gauze, place it in a dryer, and dry it at 105°C for 25 minutes so that the water content of the tea seeds is 5%, and then put the dried tea seeds into a beaker , add 1000mL of ethanol solution with a mass concentration of 75%, mix well, place in an ultrasonic oscillator, set the ultrasonic oscillation frequency to 25KHz, ultrasonic power to 150W, ultrasonic o...

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PUM

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Abstract

The invention discloses a preparation method of high-purity kallidinogenase, and belongs to the technical field of kallidinogenase extraction and preparation. The preparation method comprises the steps of mixing a tea seed extracting solution with humus sludge, filtering to obtain filter liquor, placing into tea seed bran for cooking, squeezing to prepare into tea seed bran cakes, then collecting pork pancreas and saliva, mixing, performing programmed temperature, and sequentially mixing with the tea seed bran cakes, gradually increasing amount during mixing, respectively performing three times of drip washing by using deionized water of different concentrations and sodium chloride, then collecting leacheate, adjusting a pH value through sodium hydroxide, then adding ethyl alcohol for filtering, and finally performing suction filtration and drying on obtained filter liquor to obtain the high-purity kallidinogenase. Examples show that the operation is simple, the technological level is high, not only is the specific activity enabled to reach not less than 850units / mg of protein, but also the purity is enabled to reach not less than 92 percent, and the kallidinogenase reaches state indicators and suitable for large-scale application.

Description

technical field [0001] The invention discloses a preparation method of high-purity pancreatic kininogenase, which belongs to the technical field of extraction and preparation of pancreatic kininogenase. Background technique [0002] Pancreatic kallikrein, also known as kallikrein or pancreatic kallikrein, is a type of endo-proteolytic enzyme, which is a protease isolated and purified from porcine pancreas. [0003] The pharmacological and clinical functions of pancreatic kininogenase are mainly: as a peripheral vasodilator for dilating blood vessels and improving microcirculation, activating plasmin, inhibiting phospholipase A2, reducing blood viscosity, preventing platelet aggregation, and preventing thrombosis formation etc. It is mainly used for the treatment of microcirculatory disorders, such as nephropathy caused by diabetes, peripheral neuropathy, retinopathy, fundus disease and ischemic cerebrovascular disease, and can also be used for adjuvant treatment of hyperten...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/64
CPCC12N9/6445C12Y304/21
Inventor 高玉梅张帆
Owner CHANGZHOU YAHUAN ENVIRONMENTAL PROTECTION TECH