Study On Criticality Technology Of Multi-Utilization Of The Waste Brewing Yeast Paste

Research on key technology of beer waste yeast paste multiple utilization key technology of multiple utilization of beer waste yeast paste is attached to the key agricutural program of techology hell in Jilin province(item number: 20070201,the program mainly focuses on four aspects of researches,and the main achievemens of the program is illustrated below.)1.It can be revealed by the research on the four kinds of dividing mixed and debitten of beer waste yeast methods that:(1)we have studied four kinds of beer waste yeast obtained from barley bear, adjuvant bear, rye bear, wheat bear separatively,taking the yields of clear yeast paste as parameter, analyzing the effects of concentration of yeast, screen meshing, and debittenizing on the dividing mixed and debittenizing, achieving the optimum conditions for the pretreatment. The results show that the best parameters are: concentration of the yeast: 8%, 80 screen mesh: twice, time of 5% tartrate debittenizing: 30 min. the yield of beer waste yeast can reach to 29.89%, and the electronic conductivity of the solution can reach to 4.17.(2)Comparasons on the different beer waste yeast pastes obtained from barley, adjuvant, wheat and rye tell us that :Taking the electronic conductivity as parameter, we find that the yields of beer waste yeast pastes is different in the four raw material, and adjuvant > rye > barley >wheat .The test reveals that the the level of significance of electronic conductivity of barley is less than 0.01 compared with wheat, which means that the diversity is significant. Compared with wheat, the level of significance of barley is 0.05, the signigicance levesl of barley and rye, adjuvant and wheat are all less than 0.01, which means the dervisity is extremely significant. The level of significance is not significant between adjuvant and rye, wheat and rye.(3)Analyzing the two parameters of electronic conductivity and yield of clear beer waste yeast, we have figured out the the beer waste yeast obtained from adjuvant is the optimum resource for the main experimental raw materials for the later research. Quality test after processes of dividing mixed, debitterize shows that the compounds of the beer waste yeast paste are illustrated below. Protein: 40.332%, RNA:6.539%, moisture: 73.941% . the pretreatment has been a base stone for the later research on the functions of RNA, yeast proteins, and the like.2.Researches on the amount of dissolution of RNA from beer yeast cell and the six methods of cell wall breaking tell us that:(1)Ultrasonic cell wall breaking methodWe have made the single factor experiment and the L9(34)orthogonal experiment design, and found that the relationship between factors that have effects on the amount of dissolution of RNA are ultrasonic solid-liquid ratio1(g:v)> ultrasonic power(W)> ultrasonic time(min), and the optimum parameters of technology of cell wall breaking are: ultrasonic time:5min; ultrasonic power: 500W; solid-liquid ratio1/50, amount of dissolution of RNA: 1620μg, yield of RNA: 0.181%, solid-liquid ratio1has significant effects on the dissolution of RNA.(α= 0.05)(2)Ultrasonic assistant cell wall breaking methodWe have made the single factor experiment and the L9(34)orthogonal experiment design, the results tell us that the relationship among factors that effect the dissolution of RNA is ultranic power(W)>temperature of cell wall breaking(℃)> ultrasonic time(min)> cell wall breaking time(h); the optimum conditions of ultrasonic assistant cell wall breaking method are: ultrasonic time: 20min; ultrasonic power: 600W; time of cell wall breaking: 6h, temperature of cell wall breaking: 80℃; the dissolution of RNA:652.65μg; the yield of RNA is 0.073%. the ultrasonic power has significant effects on the dissolution of RNA.(α=0.01)(3)Microwave cell wall breaking methodThrough the single factor experiment and three dimensions orthogonal polynomial regressive experimental design we have found that three dimensions orthogonal polynomial regressive equation of microwave cell wall breaking method isy=174.6-3.601Z12-113.742Z1+0.182Z3-2.798Z1Z2-0.285Z12Z2-20.829Z2(where Z1: microwave time/min, Z2: solid-liquid ratio1/g:v; Z3: microwave power/W)y=174.6-3.601Z12-113.742Z1+0.182Z3-2.798Z1Z2-0.285Z12Z2-20.829Z2,the regressive analysis tells us that the solid-liquid ratio and microwave time have significant effects on dissolution of RNA(α= 0.01 or 0.05). The value of the significant level of regressive equation is 0.01. the optimum conditions of microwave cell wall breaking method are: microwave power: 600W;microwave time: 15min; solid-liquid ratio1/45; the amount of dissolution of RNA is 1740μg, and the yield of RNA is 1.941%.(4)Microwave assistant cell wall breaking methodThrough the four factors and the four dimensions orthogonal polynomial regressive experimental design, we can contruct the four dimension linear regressive equation: y=6+0.0031Z1+1.15Z2+2.5Z3-0.106Z4-0.0011Z1Z2; (where Z1: microwave power/W; Z2:microwave time/min; Z3:time of cell wall breaking/h; Z4:temperature of cell wall breaking/℃), the regressive analysis informs us that the temperature and time of cell wall breaking have signigicant effects on the amount of dissolution of RNA.(α=0.01 or 0.05). The values of signigicant level of regressive equation is 0.01; the optimum conditions for the microwave assistant cell wall breaking are: microwave power: 800w; microwave time: 5min; temperature of cell wall breaking: 100℃;time of cell wall breaking: 3.5h; the amount of RNA dissolution: 1485.12μg, and the yield of RNA is 0.166%.(5)High voltage pulse field cell wall breaking methodWe have made the single factor experiment and the L9(34)orthogonal experiment design, and discovered that the relationship of factors that have effects on the amount of RNA dissolution is: number of pulse> solid-liquid ratio1/g:v> the electrical field strength/kv/cm. the best parameters of the high voltage pulse field cell wall breaking method are: electrical field strength: 40kv/cm, solid-liquid ratio1/50;number of pulses: 5, and the amount of RNA dissolution: 1430μg; yield of RNA: 1.603% , and the number of pulse has the significant effects on the amount of RNA dissolution.(6) high temperature salting out cell wall breaking methodThrough the L16(45)orthogonal experiment design we have discovered that: the relationship amount factors that effect the amount of RNA dissolution is: time of cell wall breaking/h> temperature of cell wall breaking /℃> concentration of yeast solution/%> concentration of NaCL solution/%. The best parameters of the high temperature salting out cell wall breaking method: time of cell wall breaking: 5h; temperature of cell wall breaking: 100℃;concentration of yeast solution: 4% ;concentration of NaCL solution: 2.257%. time and temperature of cell wall breaking have significant effects on the amount of RNA dissolution. (α=0.05)(7)The comparason among the six methods of cell wall breaking reveals that taking the amount of RNA dissolution from the beer waste yeast cell as the parameter, we can clearly see that under the best conditions of the respective method, t he high temperature salting out cell wall breaking method is the optimum(2.257%); taking the time of cell wall breaking under the same valume solution as parameter, the high voltage pulse field cell wall breaking method will cost the least time(12min).3.From the researches on the separating technologies of beer yeast protein and RNA, we found that:(1)The separating technologies of beer yeast protein and RNA. The results showed that: the optimum technology with isoelectric point precipitation RNA(pH2.7,4℃settlement 6h,centrifugate 20min with 3000r/min,in the process parameters, the RNA precipitation in the mass fraction of 0.211mg,82.176% purity);the optimum technology with ethanol precipitation(pH2.7, 4℃settlement 9h, adding ethano l20mL, centrifugate 20min with3000r/min, in the process parameters, the precipitation RNA mass fraction of 0.697mg, a purity of 72.471%).(2)The separating technologies of beer yeast protein results showed that: the isoelectric point of precipitation (pH2.7, settlement 7h, centrifugate 10min with 4200r/min, in the process parameters, the protein precipitation in the mass fraction of 33.08%, the heating aggregation(degeneration 15min, In the process parameters, the precipitation protein mass fraction of 25.38%), the isoelectric point and the heating aggregation(degeneration15min).(3)Comparition of the researches on the separating technologies of beer yeast protein and RNA, we found that: adopt the ethanol method to separate RNA; adopt the the isoelectric point and the heating aggregation to separate protein.4.Bakers' yeast proteinum enzymolysis's engineering engineering research, discover :Bakers' yeast proteinum degree of hydrolysis(DH optimum)is measurement indicatrix, in bakers' yeas proteinum density/%, proteolytic ferment concentration/%, enzymolysis time/℃, pH 4 monofactorial's empirical study foundation, though quaternionic linear regression direct cross experiment contrivance specific enforce and analysis know:(1)Construction 2 bakers' yeast proteinum control enzymolysis engineering regression model: y中性酶=119.9.4+0.420Z1+1.575Z2+0.533Z3-20.384Z4 y木瓜酶=4.982+1.239Z1+1.868Z2+0.889Z3-7.914Z4 (where Z1:bakers' yeast proteinum concentration /%, Z2: proteolytic ferment en zyme concentration /%, Z3: enzymolysis temperature/℃, Z4 :enzymolysis pH。)(2)For 2 regression model march regression coefficient analysis, regression equation significance test, partial regression coefficient test ,we knows:with dispase regression model enzymolysis's regression coefficient significance level is 0.001; regression equationsignificance level P=0.004; multiple correlation coefficient R=0.948, multiplicitas determination coefficient R2=0.899, indicate about 94% degree of hydrolysischange can use model interpret; regard topapaya proteolytic ferment regression model, 4 prediction Variable'sregression coefficient are all quiet; regression equation predominance level chalk P=0.052; multiple correlation coefficient R=0.865, multiplicitas determination coefficient R2=0.748, indicate that about 86% degree of hydrolysis alleosis can use model explain.(3)The best parameters of the two enzymes are showed: the neutral proteinase(concentration of the beer yeast protein solution1.13%, concentration of the proteinase([E]/[S])5.02%, temperature 54.3℃, and the pH 5.4, the results have been checked through three times of demonstration test, and the degree of hydrolyses have all reached to above 44.71%); the chaenomelis proteinase(concentration of the beer yeast protein solution1.34%, concentration of the proteinase([E]/[S])6.35%, temperature57.5℃, and the pH 5.4, the results have been checked through three times of demonstration test, and the degree of hydrolyses have all reached to above 26.89%). In conclusion, the regression models have great significance in guiding the large scale engineering product assembly line.