高压脉冲电场对酵母活性变化与致死双重效应研究
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摘要
为探究高压脉冲处理后酿酒酵母在工业生产领域的实际应用,以酿酒酵母为材料,研究高压脉冲电场处理后酿酒酵母活性变化和致死双重效应。在控制电场强度为唯一变量,其他影响因素不变(脉宽、频率、作用时间、脉冲个数等)的前提下对酵母悬浮液进行PEF处理,测定PEF处理后酿酒酵母的存活率和浓度及酵母活性的变化。结果表明,在电场强度1~30 k V·cm-1范围内,酵母细胞死亡比例随着电场强度的升高而增加,非特异性内酯酶活性增强的酵母细胞比例随着电场强度的升高而减少。当电场强度较低时,非特异性内酯酶活性增加的酵母比例高于死亡酵母细胞比例;电场强度大于10 k V·cm-1时,死亡酵母细胞比例高于非特异性内酯酶活性增加的酵母细胞比例;PEF处理过程中出现的亚致死酵母细胞比例在10 k V·cm-1时达到最大值。低电场PEF处理可以提高酵母细胞活性和发酵能力且酵母死亡比例较低;高电场PEF处理对酿酒酵母具有灭活效果。本研究结果对酿酒酵母的工业应用具有指导意义。
In order to explore the practical application of Saccharomyces cerevisiae in industrial production after high voltage pulse treatment,Saccharomyces cerevisiae was used as material to investigate the effect of high voltage pulsed electric field(PEF) treatment on yeast activity and lethal. The suspension of Saccharomyces cerevisiae was subjected to PEF treatment,where the field intensity was the only variable and other influencing factors(pulse width,frequency,time of action,number of pulses,etc.) were kept constant. The survival rate,the concentration of Saccharomyces cerevisiae and the change of yeast activity after PEF treatment were measured. The results showed that the proportion of yeast cell death were positively correlated with the electric field intensity,and the ratio of yeast cells with non-specific lactone activity increased were nagative correlation with electric field intensity was the range of 1 ~ 30 k V·cm-1. When the electric field intensity was low,the ratio of yeast with non-specific lactone activity increased was higher than that the ratio of dead yeast cells. When the electric field intensity was greater than 10 k V·cm-1,the ratio of dead yeast cells was higher than those non-specific lactone activity increased yeast cells. The proportion of sub-lethal yeast cells in PEF treatment reached a maximum at 10 k V·cm-1. Low-electric-field PEF treatment can improve the activity of yeast cells and the ability of fermentation,at the same time,the ratio of yeast death was low; while high-energy PEF treatment had the inactivation effect on Saccharomyces cerevisiae. The results of this study have significance for the industrial application of Saccharomyces cerevisiae.
In order to explore the practical application of Saccharomyces cerevisiae in industrial production after high voltage pulse treatment,Saccharomyces cerevisiae was used as material to investigate the effect of high voltage pulsed electric field(PEF) treatment on yeast activity and lethal. The suspension of Saccharomyces cerevisiae was subjected to PEF treatment,where the field intensity was the only variable and other influencing factors(pulse width,frequency,time of action,number of pulses,etc.) were kept constant. The survival rate,the concentration of Saccharomyces cerevisiae and the change of yeast activity after PEF treatment were measured. The results showed that the proportion of yeast cell death were positively correlated with the electric field intensity,and the ratio of yeast cells with non-specific lactone activity increased were nagative correlation with electric field intensity was the range of 1 ~ 30 k V·cm-1. When the electric field intensity was low,the ratio of yeast with non-specific lactone activity increased was higher than that the ratio of dead yeast cells. When the electric field intensity was greater than 10 k V·cm-1,the ratio of dead yeast cells was higher than those non-specific lactone activity increased yeast cells. The proportion of sub-lethal yeast cells in PEF treatment reached a maximum at 10 k V·cm-1. Low-electric-field PEF treatment can improve the activity of yeast cells and the ability of fermentation,at the same time,the ratio of yeast death was low; while high-energy PEF treatment had the inactivation effect on Saccharomyces cerevisiae. The results of this study have significance for the industrial application of Saccharomyces cerevisiae.
引文
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[2]Klomklieng W,Prateepasen A.Molasses fermentation to ethanol by Saccharomycescerevisiae M30 using low ultrasonic frequency stimulation[J].Asia-Pacific Journal of Science and Technology,2017,17(6):950-957
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[4]Huang K,Tian H,Gai L,Wang J.A review of kinetic models for inactivating microorganisms and enzymes by pulsed electric field processing[J].Journal of Food Engineering,2012,111(2):191-207
[5]Li Y Q,Tian W L,Mo H Z,Zhang Y L,Zhao X Z.Effects of pulsed electric field processing on quality characteristics and microbial inactivation of soymilk[J].Food&Bioprocess Technology,2013,6(8):1907-1916
[6]Huang K,Jiang T,Wang W,Gai L,Wang J.A comparison of pulsed electric field resistance for three microorganisms with different biological factors in grape juice via numerical simulation[J].Food&Bioprocess Technology,2014,7(7):1981-1995
[7]Marsellés-Fontanet A R,Puig-Pujol A,Olmos P,Mínguez_Sanz S,Martín-Belloso O.A comparison of the effects of pulsed electric field and thermal treatments on grape juice[J].Food&Bioprocess Technology,2013,6(4):978-987
[8]Martínez J M,Guillermo C,lvarez L,Raso J.Release of mannoproteins during Saccharomyces cerevisiae autolysis induced by pulsed electric field[J].Frontiers in Microbiology,2016,7(19778):1435
[9]Min S,Jin Z T,Zhang Q H.Commercial scale pulsed electric field processing of tomato juice[J].Journal of Agricultural&Food Chemistry,2003,51(11):3338-3344
[10]刘珂舟,袁夏冰,刘露,杨勇,韩其国,应南娇.高压脉冲电场对大肠杆菌灭菌条件的优化[J].核农学报,2015,29(8):1566-1571
[11]应南娇,付荣,杨勇,章娟.高压脉冲电场对大肠杆菌噬菌体灭活效果初探[J].核农学报,2014,28(8):1440-1445
[12]Mattar J R,Turk M F,Nonus M,Lebovka N I,Zakhem H E,Vorobiev E.S.cerevisiae fermentation activity after moderate pulsed electric field pre-treatments[J].Bioelectrochemistry,2015,103:92-97
[13]Zou T,Liu X,Ding S,Xing J.Evaluation of sperm mitochondrial function using rh123/PI dual fluorescent staining in asthenospermia and oligoasthenozoospermia[J].Journal of Biomedical Research,2010,24(5):404-410
[14]Pina-Pérez M C,Rodrigo D,López A M.Sub-lethal damage in Cronobacter sakazakii subsp.sakazakii,cells after different pulsed electric field treatments in infant formula milk[J].Food Control,2009,20(12):1145-1150
[15]Somolinos M,Ma1as P,Condón S,Pagan R,Garcia D.Recovery of Saccharomyces cerevisiae sublethally injured cells after pulsed electric fields[J].International Journal of Food Microbiology,2008,125(3):352-356
[16]陈晨,赵伟,杨瑞金,顾艳洁.PEF作用下亚致死损伤酿酒酵母修复条件的初步研究[J].食品工业科技,2014,35(21):49-53
[17]李静,廖小军,钟葵,张燕.高压脉冲电场对酿酒酵母钝化机理研究[J].农业工程学报,2011,27(4):355-360
[18]顾艳洁,赵伟,杨瑞金,闫文.高压脉冲电场作用下亚致死酵母的存在与检测[J].食品与发酵工业,2012,38(7):90-93
[19]Zeng X A,Han Z,Zi Z H.Effects of pulsed electric field treatments on quality of peanut oil[J].Food Control,2010,21(5):611-614
[20]Harrison S L,Barbosacanovas G V,Swanson B G.Saccharomyces cerevisiae structural changes induced by pulsed electric field treatment[J].LWT-Food Science and Technology,1997,30(3):236-240
[21]廖小军.高压脉冲电场系统设计及其杀菌灭酶效果与对苹果汁品质影响研究[D].北京:中国农业大学,2004
[22]Aronsson K,R9nner U,Borch E.Inactivation of Escherichia coli,Listeria innocua and Saccharomyces cerevisiae in relation to membrane permeabilization and subsequent leakage of intracellular compounds due to pulsed electric field processing[J].International Journal o Food Microbiology,2005,99(1):19-32
[23]Zakhem H E,LanoiselléJ L,Lebovka N I,Nonus M,Vorobiev E.Behavior of yeast cells in aqueous suspension affected by pulsed electric field[J].Journal of Colloid&Interface Science,2006,300(2):553-563
[24]Ohshima T,Sato M.Bacterial sterilization and intracellular protein release by a pulsed electric field[J].Advances In Biochemica Engineering/Biotechnology,2004,90(1):113-133
[25]El Zakhem H,LanoiselléJ L,Lebovka N I,Vorobiev E.The early stages of Saccharomyces cerevisiae yeast suspensions damage in moderate pulsed electric fields[J].Colloids&Surfaces BBiointerfaces,2006,47(2):189-197
[26]Zhao W,Yang R,Gu Y,Li C.Effects of pulsed electric fields on cytomembrane lipids and intracellular nucleic acids of Saccharomyces cerevisiae[J].Food Control,2014,39(1):204-213
[27]Marx G,Moody A,Bermúdezaguirre D.A comparative study on the structure of Saccharomyces cerevisiae under nonthermal technologies:high hydrostatic pressure,pulsed electric fields and thermosonication[J].International Journal of Food Microbiology,2011,151(3):327-337
[28]Rieder A,Schwartz T,Sch9n-H9lz K,Marten S M,SüJ,Gusbeth C,Kohnen W,Swoboda W,Obst U,Frey W.Molecular monitoring of inactivation efficiencies of bacteria during pulsed electric field treatment of clinical wastewater[J].Journal of Applied Microbiology,2008,105(6):2035-2045
[29]顾艳洁.高压脉冲电场下亚致死酵母的产生及进一步灭活[D].无锡:江南大学,2013
[30]Araújo O Q F,Oliveira A A C,Torres C C O,Rocha-le2o M H,Margarit I C P,Coelho M A Z.Glucose uptake in electrically stimulated cultures of Saccharomyces cerevisiae[C]//Copenhagen:Proceedings of European Congress of Chemical Engineering(ECCE-6)Copenhagen,2007:1-9
[31]Nakanishi K,Tokuda H,Soga T,Yoshinaga T,Takeda M.Effect of electric current on growth and alcohol production by yeast cells[J].Journal of Fermentation&Bioengineering,1998,85(2):250-253
[2]Klomklieng W,Prateepasen A.Molasses fermentation to ethanol by Saccharomycescerevisiae M30 using low ultrasonic frequency stimulation[J].Asia-Pacific Journal of Science and Technology,2017,17(6):950-957
[3]Muniz J B,Marcelino M,Motta M D,Schuler A,Motta A D.Influence of static magnetic fields on S.cerevisae biomass growth[J].Brazilian Archives of Biology and Technology,2007,50(3):515-520
[4]Huang K,Tian H,Gai L,Wang J.A review of kinetic models for inactivating microorganisms and enzymes by pulsed electric field processing[J].Journal of Food Engineering,2012,111(2):191-207
[5]Li Y Q,Tian W L,Mo H Z,Zhang Y L,Zhao X Z.Effects of pulsed electric field processing on quality characteristics and microbial inactivation of soymilk[J].Food&Bioprocess Technology,2013,6(8):1907-1916
[6]Huang K,Jiang T,Wang W,Gai L,Wang J.A comparison of pulsed electric field resistance for three microorganisms with different biological factors in grape juice via numerical simulation[J].Food&Bioprocess Technology,2014,7(7):1981-1995
[7]Marsellés-Fontanet A R,Puig-Pujol A,Olmos P,Mínguez_Sanz S,Martín-Belloso O.A comparison of the effects of pulsed electric field and thermal treatments on grape juice[J].Food&Bioprocess Technology,2013,6(4):978-987
[8]Martínez J M,Guillermo C,lvarez L,Raso J.Release of mannoproteins during Saccharomyces cerevisiae autolysis induced by pulsed electric field[J].Frontiers in Microbiology,2016,7(19778):1435
[9]Min S,Jin Z T,Zhang Q H.Commercial scale pulsed electric field processing of tomato juice[J].Journal of Agricultural&Food Chemistry,2003,51(11):3338-3344
[10]刘珂舟,袁夏冰,刘露,杨勇,韩其国,应南娇.高压脉冲电场对大肠杆菌灭菌条件的优化[J].核农学报,2015,29(8):1566-1571
[11]应南娇,付荣,杨勇,章娟.高压脉冲电场对大肠杆菌噬菌体灭活效果初探[J].核农学报,2014,28(8):1440-1445
[12]Mattar J R,Turk M F,Nonus M,Lebovka N I,Zakhem H E,Vorobiev E.S.cerevisiae fermentation activity after moderate pulsed electric field pre-treatments[J].Bioelectrochemistry,2015,103:92-97
[13]Zou T,Liu X,Ding S,Xing J.Evaluation of sperm mitochondrial function using rh123/PI dual fluorescent staining in asthenospermia and oligoasthenozoospermia[J].Journal of Biomedical Research,2010,24(5):404-410
[14]Pina-Pérez M C,Rodrigo D,López A M.Sub-lethal damage in Cronobacter sakazakii subsp.sakazakii,cells after different pulsed electric field treatments in infant formula milk[J].Food Control,2009,20(12):1145-1150
[15]Somolinos M,Ma1as P,Condón S,Pagan R,Garcia D.Recovery of Saccharomyces cerevisiae sublethally injured cells after pulsed electric fields[J].International Journal of Food Microbiology,2008,125(3):352-356
[16]陈晨,赵伟,杨瑞金,顾艳洁.PEF作用下亚致死损伤酿酒酵母修复条件的初步研究[J].食品工业科技,2014,35(21):49-53
[17]李静,廖小军,钟葵,张燕.高压脉冲电场对酿酒酵母钝化机理研究[J].农业工程学报,2011,27(4):355-360
[18]顾艳洁,赵伟,杨瑞金,闫文.高压脉冲电场作用下亚致死酵母的存在与检测[J].食品与发酵工业,2012,38(7):90-93
[19]Zeng X A,Han Z,Zi Z H.Effects of pulsed electric field treatments on quality of peanut oil[J].Food Control,2010,21(5):611-614
[20]Harrison S L,Barbosacanovas G V,Swanson B G.Saccharomyces cerevisiae structural changes induced by pulsed electric field treatment[J].LWT-Food Science and Technology,1997,30(3):236-240
[21]廖小军.高压脉冲电场系统设计及其杀菌灭酶效果与对苹果汁品质影响研究[D].北京:中国农业大学,2004
[22]Aronsson K,R9nner U,Borch E.Inactivation of Escherichia coli,Listeria innocua and Saccharomyces cerevisiae in relation to membrane permeabilization and subsequent leakage of intracellular compounds due to pulsed electric field processing[J].International Journal o Food Microbiology,2005,99(1):19-32
[23]Zakhem H E,LanoiselléJ L,Lebovka N I,Nonus M,Vorobiev E.Behavior of yeast cells in aqueous suspension affected by pulsed electric field[J].Journal of Colloid&Interface Science,2006,300(2):553-563
[24]Ohshima T,Sato M.Bacterial sterilization and intracellular protein release by a pulsed electric field[J].Advances In Biochemica Engineering/Biotechnology,2004,90(1):113-133
[25]El Zakhem H,LanoiselléJ L,Lebovka N I,Vorobiev E.The early stages of Saccharomyces cerevisiae yeast suspensions damage in moderate pulsed electric fields[J].Colloids&Surfaces BBiointerfaces,2006,47(2):189-197
[26]Zhao W,Yang R,Gu Y,Li C.Effects of pulsed electric fields on cytomembrane lipids and intracellular nucleic acids of Saccharomyces cerevisiae[J].Food Control,2014,39(1):204-213
[27]Marx G,Moody A,Bermúdezaguirre D.A comparative study on the structure of Saccharomyces cerevisiae under nonthermal technologies:high hydrostatic pressure,pulsed electric fields and thermosonication[J].International Journal of Food Microbiology,2011,151(3):327-337
[28]Rieder A,Schwartz T,Sch9n-H9lz K,Marten S M,SüJ,Gusbeth C,Kohnen W,Swoboda W,Obst U,Frey W.Molecular monitoring of inactivation efficiencies of bacteria during pulsed electric field treatment of clinical wastewater[J].Journal of Applied Microbiology,2008,105(6):2035-2045
[29]顾艳洁.高压脉冲电场下亚致死酵母的产生及进一步灭活[D].无锡:江南大学,2013
[30]Araújo O Q F,Oliveira A A C,Torres C C O,Rocha-le2o M H,Margarit I C P,Coelho M A Z.Glucose uptake in electrically stimulated cultures of Saccharomyces cerevisiae[C]//Copenhagen:Proceedings of European Congress of Chemical Engineering(ECCE-6)Copenhagen,2007:1-9
[31]Nakanishi K,Tokuda H,Soga T,Yoshinaga T,Takeda M.Effect of electric current on growth and alcohol production by yeast cells[J].Journal of Fermentation&Bioengineering,1998,85(2):250-253