白酒固液复合发酵模式及动力学研究
摘要
白酒是我国传统的饮料，虽然生产历史已经上千年，却仍然保持着古老的生产模式。粮食消耗量较大，生产过程大部分还是定性的，未进行定量描述，这对白酒生产的发展是不利的。鉴于此，本文针对浓香型白酒生产新模式开展了如下研究工作：
①主发酵期各关键因素与生产的关系
主要内容有培养温度对大曲制备的影响；入窖温度对粮耗及发酵升温的影响；大曲储存时间对自身生化性质及生产的影响；入窖酸度对粮耗的影响，及其他几种因素之间的相互关系。
通过对大曲的培养，得出大曲的内部温度变化的趋势。
入窖温度在17℃以上时，粮耗随温度的升高而升高．并且发酵的升温时间随入窖温度的升高而缩短．
大曲在储存6个月时，酵母数达到最高值约8×10~5个/g；细菌数降到接近最低值约0．8x10~5个/g；水分降到12%；酸度为0．86mmol/10g，接近最佳1mmol/10g；液化力为1.4g/g.h，发酵力为2.1g/g．h，糖化力为1．5g/g．h，都在最高水平上；因此储存6个月是适宜的。
入窖酸度的升高可引起粮耗的增加。
对淀粉浓度，酵母数，温度，酸度及产酒量等因素进行回归分析，发现五种因素相互之间存在较大相关；相对因变量产酒量，淀粉浓度和品温是显著因素。
②发酵动力学方程的建立
根据对微生物生长，产物形成，底物消耗的分析，建立发酵动力学方程
③酯化最佳条件的选择及酯化本初动力学的推导
对影响质量的四大酯类物质的生物合成过程进行了研究，得出最佳生成温度是35℃，最佳酸醇比是1：5-7(随酸不同而异)。在最佳条件研究的基础上，对己酸酯化动力学方程进行了推导，获得己酸在大曲催化下的本初反应动力学方程：
④新模式的建立
在以上工作的基础上，对发酵模式进行了探索，用黄水混合物进行了固液复合发酵实验，在新的发酵模式下，代表质量的总酸总酯含量同比增长116．3%和38．55％，产量同比增长20．24％。
Distillate spirits is a kind of Chinese traditional beverage.Although it has a history of thousand years,it still keeps its age-old productive model with great (?) consumption,qualitative but not quantitative production process (?),all of which is adverse for development of distillate spirit production.(?) on such a status,this paper carried out the following study in (?) to new production mode of distillate spirit:
①The relationship between key factors of main fermentation and (?)The main content includes:the effect of (?) production; (?) grain (?) temperature; the effects of Daqu depot time on (?) nature and (?); the effects of putting in acidity on grain consumption; the relationship of other important factors.
The inside temperature change tendency of Daqu can be obtained from its cultivation,and regression analysis was conducted.
When putting in temperature was above 17℃,the grain consumption rised with rising putting in temperature.The heatingup time was shortened with putting in temperature raises.
When shelf life of Daqu was 6 months,the number of yeast achieved the crest value of about 8×10~5/g;the bacteria number approximated the minimum of about 0.8×10~5/g;the water content droped to 12%;the acidity was 0.86mmol/10g,close to the optimum 1mmol/10g;the liquefy capacity was 1.4g/g.h,the fermentation capacity was 2.1g/g.h,and the saccharogenic power is 1.5g/g.h,all of them fit best; so 6 months of shelf life is eligible.
The rise of putting in acidity can cause the rise of grain consumption.
Regression analysis for the factors of starch concentration,yeast number,temperature,acidity and spirits output was conducted.The result showed that correlation among the five factors was great; starch concentration and temperature are significant factor.
②The foundation of fermentative kinetic equation
Fermentative kinetic equation can be founded on the basis of analysis on microbiological growth,productive formation and consumption of substrate.
③The selection of optimum condition and deduction on initial kinetic equation about estrification
Biosynthesis process of four kinds of esters that can affect the quality was studied.The results showed that the optimum temperature is 35℃,the optimum ratio of acerbity and alcohol is 1:5-7.Based on the optimum condition study, the kinetic equation of hexylic acid esterification was deducted, and its initial kinetic equation catalyzed by Daqu is obtained :
④The foundation of new productive mode
Based on the above operation,new fermentative mode was studied.The experiment was carried out with mixture of yellow water.With the new fermentative mode,total acerbity increased 116.3% and total ester 38.55%,and the output 20.24%.
①主发酵期各关键因素与生产的关系
主要内容有培养温度对大曲制备的影响；入窖温度对粮耗及发酵升温的影响；大曲储存时间对自身生化性质及生产的影响；入窖酸度对粮耗的影响，及其他几种因素之间的相互关系。
通过对大曲的培养，得出大曲的内部温度变化的趋势。
入窖温度在17℃以上时，粮耗随温度的升高而升高．并且发酵的升温时间随入窖温度的升高而缩短．
大曲在储存6个月时，酵母数达到最高值约8×10~5个/g；细菌数降到接近最低值约0．8x10~5个/g；水分降到12%；酸度为0．86mmol/10g，接近最佳1mmol/10g；液化力为1.4g/g.h，发酵力为2.1g/g．h，糖化力为1．5g/g．h，都在最高水平上；因此储存6个月是适宜的。
入窖酸度的升高可引起粮耗的增加。
对淀粉浓度，酵母数，温度，酸度及产酒量等因素进行回归分析，发现五种因素相互之间存在较大相关；相对因变量产酒量，淀粉浓度和品温是显著因素。
②发酵动力学方程的建立
根据对微生物生长，产物形成，底物消耗的分析，建立发酵动力学方程
③酯化最佳条件的选择及酯化本初动力学的推导
对影响质量的四大酯类物质的生物合成过程进行了研究，得出最佳生成温度是35℃，最佳酸醇比是1：5-7(随酸不同而异)。在最佳条件研究的基础上，对己酸酯化动力学方程进行了推导，获得己酸在大曲催化下的本初反应动力学方程：
④新模式的建立
在以上工作的基础上，对发酵模式进行了探索，用黄水混合物进行了固液复合发酵实验，在新的发酵模式下，代表质量的总酸总酯含量同比增长116．3%和38．55％，产量同比增长20．24％。
Distillate spirits is a kind of Chinese traditional beverage.Although it has a history of thousand years,it still keeps its age-old productive model with great (?) consumption,qualitative but not quantitative production process (?),all of which is adverse for development of distillate spirit production.(?) on such a status,this paper carried out the following study in (?) to new production mode of distillate spirit:
①The relationship between key factors of main fermentation and (?)The main content includes:the effect of (?) production; (?) grain (?) temperature; the effects of Daqu depot time on (?) nature and (?); the effects of putting in acidity on grain consumption; the relationship of other important factors.
The inside temperature change tendency of Daqu can be obtained from its cultivation,and regression analysis was conducted.
When putting in temperature was above 17℃,the grain consumption rised with rising putting in temperature.The heatingup time was shortened with putting in temperature raises.
When shelf life of Daqu was 6 months,the number of yeast achieved the crest value of about 8×10~5/g;the bacteria number approximated the minimum of about 0.8×10~5/g;the water content droped to 12%;the acidity was 0.86mmol/10g,close to the optimum 1mmol/10g;the liquefy capacity was 1.4g/g.h,the fermentation capacity was 2.1g/g.h,and the saccharogenic power is 1.5g/g.h,all of them fit best; so 6 months of shelf life is eligible.
The rise of putting in acidity can cause the rise of grain consumption.
Regression analysis for the factors of starch concentration,yeast number,temperature,acidity and spirits output was conducted.The result showed that correlation among the five factors was great; starch concentration and temperature are significant factor.
②The foundation of fermentative kinetic equation
Fermentative kinetic equation can be founded on the basis of analysis on microbiological growth,productive formation and consumption of substrate.
③The selection of optimum condition and deduction on initial kinetic equation about estrification
Biosynthesis process of four kinds of esters that can affect the quality was studied.The results showed that the optimum temperature is 35℃,the optimum ratio of acerbity and alcohol is 1:5-7.Based on the optimum condition study, the kinetic equation of hexylic acid esterification was deducted, and its initial kinetic equation catalyzed by Daqu is obtained :
④The foundation of new productive mode
Based on the above operation,new fermentative mode was studied.The experiment was carried out with mixture of yellow water.With the new fermentative mode,total acerbity increased 116.3% and total ester 38.55%,and the output 20.24%.
引文
[1] 沈怡方主编．白酒生成技术全书[M]．北京：中国轻工业出版社，1998：1-10．
[2] 沈怡方．中国新型白酒生产技术的发展[J]．酿酒科技，(2000)04：23-25．
[3] 曾祖训,新型白酒的技术进步[J]．酿酒科技，(2000)01：20-21．
[4] 沈怡方．白酒风味质量形成的主要原冈[J]．酿酒科技，(2005)1：30-34．
[5] 无锡轻工业学院等．酒精与白酒工艺学[M]．北京：轻工业出版社，1983：1-5．
[6] Alexander Dimitrov Kroumov,Aparecido Nivaldo M'odenes,Maicon C de Araujo Tait Development of new unstructured model for simultaneous saccharificationand fermentation of starch to ethanol by recombinant strain[J].Biochemical Engineering Journal 28(2006):243-255.
[7] S.Siva Kesava,T.Panda & S.K.Rakshit.Production of Ethanol by Immobilised Whole Cells of Zymomonas mobilis in an Expanded Bed[J].Bioreactor Process Biochemistry,Vol31,No.5:449-456.
[8] B.Prasad I.M.Mishra.ON THE KINETICS AND EFFECTIVENESS OF IMMOBILIZED WHOLE-CELL BATCH CULTURES [J].Bioresource Technology 53 (1995):269-275.
[9] Claudio Rossi,Marcello Porcelli,Chiara Mocenni,Nadia Marchettini,Steven Loiselle,Simone Bastianoni.A modelling approach for the analysis of xylose-ethanol[J].Bioconversion Ecological Modelling 113 (1998): 157-162.
[10] Muenduen Phisalaphong,Nuttapan Srirattana,Wiwut Tanthapanichakoon.Mathematical model of direct ethanol production from starch in Immobilized recombinant yeast culture [J].Biochemical Engineering Journal 21 (2004):93-101.
[11] Muenduen Phisalaphong,Nuttapan Srirattana,Wiwut Tanthapanichakoon.Mathematical modeling to investigate temperature effect on kineticparameters of ethanol fermentation[J].Biochemical Engineering Journal 28 (2006):36-43.
[12] Elmer Ccopa Riveraa,Aline C.Costa a,Daniel I.P.Atala b,Francisco Maugerib,Maria R.Wolf Maciel a,Rubens Maciel Filhoa.Evaluation of optimization techniques for parameter estimationApplication to ethanol fermentation considering the effect of temperature[J].Process Biochemistry 41 (2006): 1682-1687.
[13] L.E.Romero,D.Cantero.Mathematical Model for Liquid-Gas Equilibrium in Acetic Acid Fermentations[J].BIOTECHNOLOGY AND BIOENGINEERING,VOL.59,NO.3,AUGUST 5,1998.
[14] 戚以政，汗叔雄编著．生化反应动力学与反应器[M]北京：化学工业出版社，1999．
[15] Salman Zafar.Mohammad Owais Ethanol production from crude whey by Kluyveromyces marxianus [J].Biochemical Engineering Journal,27 (2006) 295-298.
[16] Morteza Khanahmadia,David A.Mitchellb,Masoud Beheshtic,Reza Roostaazadc,Luis R.Sanchezd.Continuous solid-state fermentation as affected by substrate flowpattern[J].Chemical Engineering Science 61(2006): 2675-2687.
[17] Luong J.H.T.Generalization of Monod kinetics for analysis of growth data with substrate inhibition.Biotechnol Bioeng [J].1987,29,February:242-248.
[18] Fumiko Tanakaa,Akira Andoa,Toshihide Nakamuraa,Hiroshi Takagib,Jun Shima.Functional genomic analysis of commercial baker's yeast during initialstages of model dough-fermentation[J].Food Microbiology 23 (2006) 717-728.
[19] J.P.Smits,A.Rinzema:J.Tramper,H.M.van Sonsbeek,J.C.Hage: A.Kaynak,and W.Knol.The influence of temperature on kinetics in solid-state fermentation [J].Enzyme and Microbial Technology 1998 22:50-57.
[20] Wissam Medawar,Pierre Strehaiano,Marie-Line Delia.Yeast growth: lag phase modelling in alcoholic media[J].Food Microbiology 20 (2003):527-532.
[21] Levenspiel O.The Monod equation:a revisit and a generalization to product inhibitio situations.Biotechnol.Bioeng.[J].1980,22:1671-1687.
[22] Luong J.H.T.Kinetics of ethanol inhibition in alcohol fermentation [J].Biotechnol.Bioeng.l985,27:280-285.
[23] Cysewski G.R,Wilke C.R.Membrane separation in production of alcohol fuels by fermentation.In:MeGegor W C,ed.Membrane separations in biotechnology [M].New York:Marcel-Dekker,1987:43-89.
[24] Kargupta K.,Datta S,Sanyal S.K.Analysis of the performance of a continuous membrane bioreactor with cell recycling during ethanol fermentation [J].Biochemical Engineering Journal,1998,1:31-37.
[25] Cho C.W.Hwang S.T.Continuous membrane fermentor separator for ethanol fermentation[J].J.Membr.Sc.l991,57:21-42.
[26] 张卫，虞星炬，袁权，张衍坤，张铭俊．乙醇发酵-完全细胞截留渗透汽化膜分离耦合过程[J]，膜科学与技术．1997，3(17)：42-47
[27] 山根恒夫著．苏尔馥，胡章助，译．生物反应工程[M]．上海：上海科学技术出版社．1989：176-179．
[23] Lee C.W.,Chang H.W.Kinetics of ethanolfermentations in membrane cell recycles Fermentors[J].Biotechnol Bioeng,29,l105-1112.
[29] Shekar Govindaswamy,Leland M.Vane Kinetics of growth and ethanol production on different carbon substrates using genetically engineered xylose-fermenting yeast[J].Bioresource Technology 98 (2007):677-685.
[30] 吴衍庸．红曲霉孢外酯酶催化己酸乙酯合成的研究，生物工程学报．1995,11(3):388-390
[2] 沈怡方．中国新型白酒生产技术的发展[J]．酿酒科技，(2000)04：23-25．
[3] 曾祖训,新型白酒的技术进步[J]．酿酒科技，(2000)01：20-21．
[4] 沈怡方．白酒风味质量形成的主要原冈[J]．酿酒科技，(2005)1：30-34．
[5] 无锡轻工业学院等．酒精与白酒工艺学[M]．北京：轻工业出版社，1983：1-5．
[6] Alexander Dimitrov Kroumov,Aparecido Nivaldo M'odenes,Maicon C de Araujo Tait Development of new unstructured model for simultaneous saccharificationand fermentation of starch to ethanol by recombinant strain[J].Biochemical Engineering Journal 28(2006):243-255.
[7] S.Siva Kesava,T.Panda & S.K.Rakshit.Production of Ethanol by Immobilised Whole Cells of Zymomonas mobilis in an Expanded Bed[J].Bioreactor Process Biochemistry,Vol31,No.5:449-456.
[8] B.Prasad I.M.Mishra.ON THE KINETICS AND EFFECTIVENESS OF IMMOBILIZED WHOLE-CELL BATCH CULTURES [J].Bioresource Technology 53 (1995):269-275.
[9] Claudio Rossi,Marcello Porcelli,Chiara Mocenni,Nadia Marchettini,Steven Loiselle,Simone Bastianoni.A modelling approach for the analysis of xylose-ethanol[J].Bioconversion Ecological Modelling 113 (1998): 157-162.
[10] Muenduen Phisalaphong,Nuttapan Srirattana,Wiwut Tanthapanichakoon.Mathematical model of direct ethanol production from starch in Immobilized recombinant yeast culture [J].Biochemical Engineering Journal 21 (2004):93-101.
[11] Muenduen Phisalaphong,Nuttapan Srirattana,Wiwut Tanthapanichakoon.Mathematical modeling to investigate temperature effect on kineticparameters of ethanol fermentation[J].Biochemical Engineering Journal 28 (2006):36-43.
[12] Elmer Ccopa Riveraa,Aline C.Costa a,Daniel I.P.Atala b,Francisco Maugerib,Maria R.Wolf Maciel a,Rubens Maciel Filhoa.Evaluation of optimization techniques for parameter estimationApplication to ethanol fermentation considering the effect of temperature[J].Process Biochemistry 41 (2006): 1682-1687.
[13] L.E.Romero,D.Cantero.Mathematical Model for Liquid-Gas Equilibrium in Acetic Acid Fermentations[J].BIOTECHNOLOGY AND BIOENGINEERING,VOL.59,NO.3,AUGUST 5,1998.
[14] 戚以政，汗叔雄编著．生化反应动力学与反应器[M]北京：化学工业出版社，1999．
[15] Salman Zafar.Mohammad Owais Ethanol production from crude whey by Kluyveromyces marxianus [J].Biochemical Engineering Journal,27 (2006) 295-298.
[16] Morteza Khanahmadia,David A.Mitchellb,Masoud Beheshtic,Reza Roostaazadc,Luis R.Sanchezd.Continuous solid-state fermentation as affected by substrate flowpattern[J].Chemical Engineering Science 61(2006): 2675-2687.
[17] Luong J.H.T.Generalization of Monod kinetics for analysis of growth data with substrate inhibition.Biotechnol Bioeng [J].1987,29,February:242-248.
[18] Fumiko Tanakaa,Akira Andoa,Toshihide Nakamuraa,Hiroshi Takagib,Jun Shima.Functional genomic analysis of commercial baker's yeast during initialstages of model dough-fermentation[J].Food Microbiology 23 (2006) 717-728.
[19] J.P.Smits,A.Rinzema:J.Tramper,H.M.van Sonsbeek,J.C.Hage: A.Kaynak,and W.Knol.The influence of temperature on kinetics in solid-state fermentation [J].Enzyme and Microbial Technology 1998 22:50-57.
[20] Wissam Medawar,Pierre Strehaiano,Marie-Line Delia.Yeast growth: lag phase modelling in alcoholic media[J].Food Microbiology 20 (2003):527-532.
[21] Levenspiel O.The Monod equation:a revisit and a generalization to product inhibitio situations.Biotechnol.Bioeng.[J].1980,22:1671-1687.
[22] Luong J.H.T.Kinetics of ethanol inhibition in alcohol fermentation [J].Biotechnol.Bioeng.l985,27:280-285.
[23] Cysewski G.R,Wilke C.R.Membrane separation in production of alcohol fuels by fermentation.In:MeGegor W C,ed.Membrane separations in biotechnology [M].New York:Marcel-Dekker,1987:43-89.
[24] Kargupta K.,Datta S,Sanyal S.K.Analysis of the performance of a continuous membrane bioreactor with cell recycling during ethanol fermentation [J].Biochemical Engineering Journal,1998,1:31-37.
[25] Cho C.W.Hwang S.T.Continuous membrane fermentor separator for ethanol fermentation[J].J.Membr.Sc.l991,57:21-42.
[26] 张卫，虞星炬，袁权，张衍坤，张铭俊．乙醇发酵-完全细胞截留渗透汽化膜分离耦合过程[J]，膜科学与技术．1997，3(17)：42-47
[27] 山根恒夫著．苏尔馥，胡章助，译．生物反应工程[M]．上海：上海科学技术出版社．1989：176-179．
[23] Lee C.W.,Chang H.W.Kinetics of ethanolfermentations in membrane cell recycles Fermentors[J].Biotechnol Bioeng,29,l105-1112.
[29] Shekar Govindaswamy,Leland M.Vane Kinetics of growth and ethanol production on different carbon substrates using genetically engineered xylose-fermenting yeast[J].Bioresource Technology 98 (2007):677-685.
[30] 吴衍庸．红曲霉孢外酯酶催化己酸乙酯合成的研究，生物工程学报．1995,11(3):388-390