The Research Of The Different Mothod Induces Mutation With T-3 The Natto Activating Enzyme Strain

The natto activating enzyme was the serine proteinase which produces in the natto bacillus fermentative process, has the good filament solubility. With the microorganism fermentation method production natto activating enzyme was the method which in the modern natto activating enzyme production first uses, therefore the people also take the natto activating enzyme fermentation to use the mold mushroom spawn the research. This experiment's goal was through the mutation breeding method, obtains one when the fermentation can have the high natto activating enzyme enzyme vigor strain. The experiment uses this laboratory preservation the natto bacillus T-3 strain to embark the strain, selects the ultraviolet ray, the microwave, the ultrasonic wave, nitroso group guanidine and the nitroso group guanidine - ultrasonic wave union mutagenesis method, through the single factor experiment, the multi-factor overlapping combination test, the revolving combination test design's method, studies each mutafacient method the mutafacient condition, And enhances the natto activating enzyme production strain through the mutagenesis to produce the enzyme ability, breeds selectively the natto activating enzyme high production strain, provides the fine mold mushroom spawn for the zymotechnics production natto activating enzyme.1.The ultraviolet radiation method induces mutation or chromosomal change the T-3 natto bacillus mutagenesis condition the research. In induces mutation or chromosomal change in the T-3 natto bacillus's experiment using the ultraviolet radiation method, as embarks the strain take the T-3 natto bacillus, studies the different ultraviolet radiation time to induce mutation or chromosomal change the effect the influence. Through the single factor experiment determination ultraviolet ray mutagenesis's time, and under this mutagenesis time's condition, the natto bacillus carries on mutafacient processing to T-3. The test result indicated: The ultraviolet ray mutagenesis's irradiation time was 240s, undergoes two round mutagenesis and screening, obtains the sudden change strain U2-5 fermentation production to obtain the natto activating enzyme vigor was 5440.37 IU/mL, enhances 8.8 % compared to the primitive strain, and heredity stable property.2.The microwave radiation method induces mutation or chromosomal change the T-3 natto bacillus mutagenesis condition the research. In induces mutation or chromosomal change in the T-3 natto bacillus's experiment using the microwave radiation method, as embarks the strain take the T-3 natto bacillus, through the single factor experiment, studies the different microwave processing power and the different microwave process time under condition microwave to T-3 natto bacillus mutagenesis effect, thus the determination microwave processing time and the power, and under this microwave mutagenesis time's condition, carry on mutafacient processing to the T-3 natto bacillus, and initially sieves using the casein plate with the rocker fermentation affination method, after screening the microwave radiation mutagenesis sudden change. The test result indicated: The microwave mutagenesis's power was 380 W, Heating accumulation time 40 s, undergoes two round mutagenesis and screening obtains the sudden change W2-1 strain, its fermentation production obtains the natto activating enzyme vigor was 5450.00 IU/mL, enhances 9 % compared to the primitive strain, and heredity stable property.3. The ultrasonic wave processing method induces mutation or chromosomal change the T-3 natto bacillus mutagenesis condition the research. In induces mutation or chromosomal change in the T-3 natto bacillus's experiment using the ultrasonic wave processing method, as embarks the strain take the T-3 natto bacillus, through single factor experimental study different ultrasonic wave power, frequency, Time under condition ultrasonic wave to T-3 natto bacillus mutagenesis effect. In the single factor experiment's foundation, uses the orthogonal test optimization mutagenesis condition, thus the determination ultrasonic wave processing power, the frequency and the time, and under this ultrasonic wave mutagenesis condition, carry on mutafacient processing to the T-3 natto bacillus, and initially sieves using the casein plate with Rocker fermentation affination method, after screening favorable balance of trade sound emission mutagenesis sudden change. The test result indicated: The ultrasonic wave processing power was 220 W, accumulation response time 25 min, frequency 70 KHz, carries on two round mutagenesis and screening to the T-3 natto bacillus obtains the sudden change C2-1 strain, its fermentation production obtains the natto activating enzyme vigor is 5585.35 IU/mL, enhances 11.7 % compared to the primitive strain, and heredity stable property.4. The nitroso group guanidine processing method induces mutation or chromosomal change the T-3 natto bacillus mutagenesis condition the research. In induces mutation or chromosomal change in the T-3 natto bacillus's experiment using the nitroso group guanidine processing method, as embarks the strain take the T-3 natto bacillus, through the single factor experiment, studies the different nitroso group guanidine process time and different nitroso group guanidine Processing density under condition nitroso group guanidine to T-3 natto bacillus mutagenesis effect, thus the determination nitroso group guanidine processing time and the density, and under this process condition, carry on mutafacient processing to the T-3 natto bacillus, initially sieves using the casein plate with the rocker fermentation affination method, screens the nitroso group guanidine mutagenesis Latter sudden change. The test result indicated: Through single factor experiment determination nitroso group guanidine process time 20 min and nitroso group guanidine processing density 0.3mg/mL, carries on two round mutagenesis and screening to the T-3 natto bacillus obtains the sudden change Y2-1 strain, its fermentation production obtains the natto activating enzyme vigor is 5740.25 IU/mL, enhances 14.8 % compared to the primitive strain.5. The nitroso group guanidine-ultrasonic wave union mutagenesis method induces mutation or chromosomal change the T-3 natto bacillus mutagenesis condition the research. In the nitroso group guanidine- ultrasonic wave union mutagenesis method induces mutation or chromosomal change in the T-3 natto bacillus's experiment, as embarks the strain take the T-3 natto bacillus, through single factor experimental study different nitroso group guanidine density, The union mutagenesis process time, the processing frequency under the condition mutagenesis processes to the T-3 natto bacillus mutagenesis effect. In the single factor experiment's foundation, uses two return orthogonal revolving combination test design, the optimization induces mutation or chromosomal change the T-3 natto bacillus's mutafacient condition jointly, and Under the optimized mutafacient condition, carries on mutafacient processing to the T-3 natto bacillus, and initially sieves using the casein plate with the rocker fermentation affination method, after screening the union mutagenesis sudden change. The test result indicated: The T-3 natto bacillus's mutafacient condition for nitroso group guanidine density 0.3 mg/mL, process time 20 min, processing frequency 70 KHz, carries on mutafacient processing to the T-3 natto bacillus, Screens sudden change F2-1, it produces the natto activating enzyme vigor in 6600.52 IU/mL, enhanced 30.0 % compared to primitive strain T-3.Draws the conclusion through the research, the ultraviolet ray, the microwave, the ultrasonic wave, nitroso group guanidine and the nitroso group guanidine - ultrasonic wave induces mutation or chromosomal change these five mutafacient method to have the good mutafacient effect jointly, the nitroso group guanidine - ultrasonic wave union mutagenesis method's effect was best, obtains sudden change with this mutafacient method to produce the natto activating enzyme the vigor most to be high was 6600.52 IU/mL, surpasses other to induce mutation or chromosomal change the method solely the mutafacient effect.