Black Trichoderma Reesei Fermentation Productiom Of T-PA And Kineticstudies

This paper summarizes the present status of thrombotic diseases, the role of t-PA mechanism, the t-PA study historyand the status quo, t-PA physical and chemical properties and biological activity, genetic engineering of the production of t-PA, people from research t-PA start with the structure, the structure-activity relationship to the principle of using recombinant DNA and protein engineering technology building a series of t-PA variant, which extended its half-life in vivo, an increase of fibrin affinity and improve the fibrinolytic The catalytic activity of plasminogen areas have greater improvement than the natural t-PA more broad application prospects. And the production of t-PA was introduced, the use of genetic engineering of high-yielding strains of Trichoderma reesei, through fermentation flask experiment, the purified liquid fermentation, separation and purification steps are: the supernatant collection system solution, after a slightly lysine affinity chromatography, received t-PA pure. And on t-PA detection method and the t-PA clinical application.This paper mainly on the production of t-PA genetically engineered bacteria Trichoderma reesei 306 for breeding, and its fermentation conditions, the fermentation medium composition and dynamic characteristics were studied.First, using genetically engineered bacteria Trichoderma reesei 306 as the starting strains isolated by screening, and then flask-screen, select a high yield of t-PA 306 (8).Secondly, the study of culture conditions on seeds through single factor experiment to determine the high-yielding strain 306 (8) the seeds of the culture time for 32 hours, after 32 hours of training on the number of seeds into the growth phase of the latter part of the initial period of stability, seed the most dynamic through the flask experiment to determine the high-yielding strain 306 (8) seed inoculation of 12%, inoculum size is too small and too much of the middle class is not good.Third, the strains on the fermentation medium conditions, the application of quantitative analysis single factor experiment, response surface experiment to determine the optimal formula for the media: 2% bran water +1.25% lactose +0.8% ammonium sulfate + 0.2 Pancreatic peptone +0.1% Tween-80.Fourth, the single factor experiment, research, culture fermentation conditions that determine the initial pH of the medium was 5.4; ventilation capacity of 150 r / min; medium volume of 30 ml, but also medium components were analyzed found that surfactant Tween 80 and the promotion of sodium acetate t-PA has to improve production. Under these conditions t-PA output up 80.72 U / ml, respectively.Fifth, the fermentation broth of initial treatment were studied to determine the salt out of the ammonium sulfate saturation of 50% and 60% by the stability of the experimental studies to determine protease should be kept in refrigerator layer.Finally, the t-PA study of the dynamics of fermentation, identified with the time, cell growth, the consumption of substrate and the product of the law .