Optimization Of Fermentation Process On A Fibrinolytic Enzyme And Primary Analysis Of The Enzyme Property

Thrombus is a kind of disease which leads to tip top death rate in clinic for the moment. Scientists of medicine vocation are always paying attention to searching of deliquescent thrombus medicament in the world. And people have been developing the novel medicine to this day.The experiment had screened two superiority strains which named HFZ-1A and HFZ-2D that can produce the higher activity fibrinolytic enzyme.Fermentation conditions, including medium, inoculation quantity, strain age, pH, OD, rpm, had been optimized.Then, purified fibrinolytic enzyme named M-1 A and M-2D,studied prorerties of enzyme and dissolution of thrombolyic. Finally,observed the acute toxicity of M-1 A and M-2D on mouse.1. Fermentation conditions of HFZ-1A and HFZ-2D were optimized through single factor test and orthogonal design study. The conclusion was as follows:(1) HFZ-1A stain: soluble starch 4%,peptone 2%, ammonium ferric citrate 0.1%,calcium phosphate 0.4%,SDS 0.1%,the inoculum size 2%,pH 7.5, fermentation time 96 h,media amount 75/500 (mL/mL),fermentation temperature 37 °C, rotate speed 150 r/min.Under these conditions,the strain's fibrinolytic enzyme activity was 581.81 IU/mL.(2) HFZ-2D stain: soluble starch2%, urea2%, ammonium ferric citrate0.4%, KH₂PO₄0.1%, PEG 0.1%,the inoculum size 4%,pH7.5, fermentation time 72h,media amount 100/500(mL/mL),fermentation temperature32°C,rotate speed 180r/min.Under these conditions,the strain's fibrinolytic enzyme activity was 693.97 IU/mL.(3) Compared to correlative study, fibrinolytic enzyme activity of HFZ-1 A and HFZ-2D strain is higher.2. After purifying, enzyme activity of M-1A reached 1032.69 IU/mL and the activity of M-2D reached 1258.14 IU/mL.The characters of these two enzymes had been studied.The optimum pH and optimum temperature for M-1A and M-2D to dissolute thrombolyic are pH7.5⁸.5 and 45°C⁵0°C,and theses two enzymes activities are not stable at high temperature and low pH, Fe³, Mn², NH₄⁺ activates the activaty of M-1 A and SO₄^2-, Zn²⁺,...