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Preparation for asparaginic acid protease inhibitors

A protease inhibitor, aspartic acid technology, applied in the field of fungal fermentation bioactive products, can solve the limitations; foreign countries have extracted aspartic acid from potatoes, tomatoes and legumes and other problems, reaching the high level of medicine Health care value, good application prospect, effect of eliminating pollution

Active Publication Date: 2008-10-01
江苏博立生物制品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the protease A inhibitors reported at home and abroad are synthesized by chemical methods, and these inhibitors are subject to considerable restrictions in application; foreign countries have extracted aspartic acid from potatoes, tomatoes and legumes. Protease inhibitors; our laboratory has mentioned two protease A inhibitors, GLPIA1 and GLPIA2, from Ganoderma lucidum fermented whole powder, and extracted a protease A inhibitor from potato tubers

Method used

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  • Preparation for asparaginic acid protease inhibitors
  • Preparation for asparaginic acid protease inhibitors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Put the Yunzhi slant into the seed medium, after cultivation, fill the flask with a liquid volume of 75mL / 250mL, and then put it into the fermentation medium according to the inoculation volume of 10%. The composition of the seeds and the fermentation medium is as described in the instructions. The initial pH was 6.0, cultured at 26°C and 170r / min for 5 days. Put the fermentation broth in a -20°C refrigerator to freeze, take it out and thaw it in a water bath at 30°C, and repeat this process 3 times; ultrasonically break the cells according to the ultrasonic time of 10min, power of 280W, temperature of 20°C, and processing capacity of 40mL, and centrifuge at 12000r / min for 15mim. The precipitate was discarded, and 65 mL of the supernatant collected after multiple batches of cell disruption and centrifugation was decolorized with macroporous resin D101 to obtain 60 mL of crude inhibitor, the concentration ratio of protein to sugar was about 0.77, and the IC for pepsin 50...

Embodiment 2

[0036] Insert the Yunzhi plate into the seed medium, after cultivation, fill the flask with a liquid volume of 75mL / 250mL, and then insert it into the fermentation medium according to the inoculation volume of 10%. The composition of the seeds and fermentation medium is as described in the instructions. The initial pH was 6.0, cultured at 26°C and 170r / min for 5 days. Place the fermentation broth in a -20°C refrigerator to freeze, take it out and thaw it in a water bath at 30°C, and repeat this process 3 times; ultrasonically break the cells according to the ultrasonic time of 10min, ultrasonic power of 280W, ultrasonic temperature of 20°C, and processing capacity of 40mL, and centrifuge at 12000r / min 15mim, discard the precipitate, and decolorize the 65mL supernatant collected after multiple batches of cell disruption and centrifugation with macroporous resin D101 to obtain a crude inhibitor. The concentration ratio of protein to sugar is about 0.93, and the IC for pepsin 50 ...

Embodiment 3

[0038] The Yunzhi plate is directly connected to the fermentation medium, and the composition of the fermentation medium is as described in the instructions. The initial pH was 6.0, cultured at 26°C and 170r / min for 5 days. Place the fermentation broth in a -20°C refrigerator to freeze, take it out and thaw it in a water bath at 30°C, and repeat this process 3 times; ultrasonically break the cells according to the ultrasonic time of 10min, ultrasonic power of 280W, ultrasonic temperature of 20°C, and processing capacity of 40mL, and centrifuge at 12000r / min 15mim, discard the precipitate, and decolorize the 65mL supernatant collected after multiple batches of cell disruption and centrifugation with macroporous resin D101 to obtain a crude inhibitor. The concentration ratio of protein to sugar is about 0.73, and the IC for pepsin 50 Value is about 1.99mg / mL, yield is about 83.4%; Get 15mL crude inhibitor and carry out DEAE52 ion-exchange chromatography, column specification is ...

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Abstract

The present invention provides a preparing method of aspartic proteinase inhibitor, belonging to technical field of fungi fermentation bioactivity product. The invention adopts Lucid Ganoderma fermentation, joining the Lucid Ganoderma in culture medium which the main component is potato, pH is adjusted by HCl, fermenting culture, collecting cells, repeated freezing and thawing, breaking cells by combining with ultrasonic, centrifugatiing in high speed, obtaining crude inhibitor by decoloring the supernatant by a macroporous resin, obtaining the purifying aspartic proteinase inhibitor by DEAE52 ion exchange chromatography, obtaining the aspartic proteinase inhibitor powder by vacuum drying. The inhibitor prepared by the method inhibits activity of aspartic proteinase, the crude inhibitor containing all kinds of fermentation products has high medical and health prices, having no pigment effect in the application by decoloring treatment, keeping security, having better application prospects in medical, food additive and insect prevention field.

Description

technical field [0001] A method for preparing an aspartic acid protease inhibitor, specifically using Yunzhi fermentation, repeated freezing and thawing combined with ultrasonic waves for cell disruption, decolorization by macroporous resin, and DEAE ion exchange chromatography to obtain relatively pure aspartic acid The invention relates to an acid protease inhibitor, which belongs to the technical field of fungal fermentation bioactive products. Background technique [0002] In China, as the "pure heat" continues to heat up, the foam stability of pure draft beer has attracted more and more attention from manufacturers and researchers. Existing studies have proved that the existence of active protease A and inactive protease A precursor will directly or indirectly destroy the foam protein of pure draft beer, thereby reducing the foam stability of pure draft beer. Protease A and its precursor The total amount of the substance determines the comprehensive ability to degrade ...

Claims

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Application Information

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IPC IPC(8): C12P1/02C12R1/645
Inventor 田亚平马亚敏刘琦
Owner 江苏博立生物制品有限公司
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