Method for preparing ganoderma sinensis streptokinase and application thereof

Abstract

The invention relates to a process for preparing purple chi thrombolytic enzyme, which comprises solid state fermentation process of purple chi, firstly, activating bacteria, evenly coating purple chimycelium on inclined medium surface to culture, secondly, culturing seeds, utilizing soya bean to be seed culture medium, inoculating activated strain which is obtained from the step one to seed culture medium shake bottle to culture, thirdly, preparing soya bean to be solid fermentation culture medium, fourthly, fermenting to produce enzyme, inoculating bacterial liquid which is obtained from the step two to solid fermentation culture medium which is obtained from the step three to ferment and culture to obtain solid fermentation material, separation and purification process of enzyme comprises firstly, preparing crude enzyme preparation, adding solid fermentate material which is obtained from the above solid state fermentation process into sodium phosphate buffer solution homogenate, stirring to prepare and centrifuging, getting supernatant fluid to be crude enzyme solution, second, purifying enzyme, doing Sephadex G-50 pillar layer chromatography for crude enzyme solution which is obtained from step one, gathering eluent portion which has enzymatic activity, utilizing CMC ion to change pillar layer chromatography, gathering eluent portion of enzymatic activity peak, and obtaining purified purple chi thrombolytic enzyme.

Description

technical field [0001] The present invention relates to a preparation method of a thrombolytic enzyme and the application of the thrombolytic enzyme, in particular to a method for preparing a thrombolytic enzyme from Zizhi lucidum and the application of the prepared thrombolytic enzyme in the preparation of medicines for treating thromboembolic diseases . Background technique [0002] Thromboembolic disease is one of the main diseases that endanger people's health and life. At present, the incidence of thromboembolic diseases caused by various reasons, such as cerebral thrombosis, myocardial infarction, coronary heart disease, and limb vascular embolism, is on the rise. Among them, There are about 15 million patients with thromboembolic diseases in the world, and their morbidity, disability and mortality are very high, seriously threatening human health and life. One of the main causes of thromboembolic diseases is due to the formation of thrombus in blood vessels, and the ...

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Problems solved by technology

[0008] Among the thrombolytic agents currently used clinically, streptokinase (SK) and urokinase (UK) are mainly used, but these two drugs have their limitations: streptokinase is prone to produce streptokinase antibodies in the human body, so there is antagonism, The dosage is difficult to control, and has obvious side effects; although urokinase has no obvious antigenicity, because urokinase is very unstable, its activity is difficult to control, and it is easy to cause massive bleeding or fail to dissolve thrombus due to inactivation

At the same time, the preparation process of these thrombolytic agents is complicated, expensive, and difficult to be widely used.

[0009] In view of the difficulty in the source of commercialized thrombolytic agents, complex production processes and other factors, the output of this type of product is limited, which directly affects the treatment of diseases. procedures, causing adverse consequences to environmental sanitation and affecting the lives of surrounding residents; at the same time, due to weather changes, urine is easy to deteriorate, which directly affects the output of production; The nattokinase yield of bacillus (Bacillus natto) is not high, and the production method of nattokinase obtained thus is also not ideal enough

All these factors affect the yield and quality of thrombolytic preparations, thus limiting their popularization and application.

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