Submerged Culture Condition For Fibrinolytic Enzyme Production, Anticoagulant And Thrombolytic Effects Of The Enzyme By Acidophilic Actinomycete701

Thromboembolic disease endangers human health each year in the world. So far,every kinds of thrombolysis drug still have some disadvantages, therefore, it is very important to develop a new thrombolysis drug with safe, effective and low-cost characteristics. Actinomycetes is a natural bacteria which can produce many kinds of bioactive constituents. Study findout some strong fibrinolytic activity constituents in its fermentation liquor, which is very important for developing a new drug of dissolving thrombus. Therefore, in this paper, an Acidophilic actinomycete701was investigated, and got some important results.An Acidophilic actinomycete701was investigated, and obtained the optimumparameters of the submerged fermentation medium and conditions. We investiaged thefermentation kinetics, and modeled the fermentation kinetics equations in shaking culture theunder the optimization culture medium and conditions. And then we used the anticoagulantand thrombolytic experiments to prove the anticoagulant and thrombolytic effects. Simulationhyperlipidemia animal experiments were used to prove the effects of blood lipid reducted andarteriosclerosis prevented the fibrinolytic enzyme, which can help to develop a newgeneration thrombolysis drug.Enzyme activity of fibrinolytic enzyme of Acidophilic actinomycete701as the index,and obtained the optimum parameters of the submerged fermentation medium and conditions.The optimum conditions were obtained by single factor experiment, the optimum nitrogensources was beef extract, the optimum metal ions were Ca²⁺and Na⁺, the optimumconcentration of millet was1%, liquid volume was30mL in250mL flask. Plackett-Burmandesign was used to evaluate the eight factors which related to fibrinolytic enzyme productivity,and four factors （initial pH, beef extract, inoculum size, liquid volume） have significanteffects on enzyme activity. Finally, the four major factors were optimized by response surfacemethodology, and the results showed that the optimal culture conditions were initial pH of5.32, beef extract of1.13%, inoculums size of7.1%and liquid volume of20mL/250mL;fibrinolytic activity increased to228.17mm²/10μL, and under the optimum conditions, theenzyme activity increased41.02%. The fermentation kinetics of Acidophilic actinomycete701submerged batch culture forfibrinolytic enzyme production was investigated. The results showed that there was arelationship between the production of fibrinolytic enzyme and actinomycete, the kineticequations for biomass growth, production of fibrinolytic enzyme and consumption of glucosewere established as follow：These kinetics models were good agreement with the experimental results.The anticoagulation and thrombolytic experiment results of Acidophilic actinomycete701showed that the fibrinolytic enzyme have good anticoagulant and thrombolytic effects,and the thrombus hydrolysis degree reached to65%when in vitro experiments. Induced miceto produce hyperlipidemia by feeding high fat feed, to prove regulating action for blood lipidof the fibrinolytic enzyme, results showed that fibrinolytic enzyme have significant effects ondecreasing the concentration of TC, TG and LDL-C in blood lipid, and rising theconcentration of HDL-C, and inhibiting the formation of atherosclerotic plaque, reducing therisk of blood clots.