混菌发酵体系中异常汉逊酵母生长抑制机制研究
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摘要
为揭示异常汉逊酵母(Hansenula anomala)早期衰亡的特征及机制,建立酿酒酵母(Saccharomycescerevisiae)与异常汉逊酵母混合发酵体系,研究酿酒酵母产酸、有氧混菌发酵、无氧添加新鲜培养基发酵、混菌发酵上清液及活、死酿酒酵母细胞添加对异常汉逊酵母生长的影响。结果表明,酵母产酸对异常汉逊酵母有抑制作用;有氧混菌和无氧添加新鲜培养基混菌发酵处理中,异常汉逊酵母的活菌数均高于无氧混菌发酵;而混菌发酵上清液添加后异常汉逊酵母的活菌数低于其纯培养;高浓度活酿酒酵母细胞的加入导致异常汉逊酵母的早期死亡,而死酿酒酵母细胞对异常汉逊酵母无抑制作用。因此,异常汉逊酵母早期衰亡,一方面是由酿酒酵母产酸及营养竞争造成,另一方面酿酒酵母的代谢产物及高密度活酿酒酵母细胞也会有促进作用。上述结果有助于加深对非酿酒酵母在果酒发酵中早于酿酒酵母衰亡的理解。
In order to reveal the characteristics and mechanism of early death of Hansenula anomala,this paper established a mixed fermentation system for Saccharomyces cerevisiae and H. anomala; and studied the effects of acids produced by S. cerevisiae,aerobic fermentation,mixed-culture in fresh medium without oxygen,mixed-culture supernatant,viable or dead S. cerevisiae cells on the growth of H. anomala. The results showed that the growth of H.anomala was inhibited by acid produced by yeast. In the fermentation treatments of aerobic mixed bacteria and anaerobic add fresh medium,the viable counts of H. anomala were all higher than that of anaerobic mixture fermentation; while the viable count of H. anomala in mixed-culture supernatant was lower after adding fresh medium.The presence of viable S. cerevisiae cells at high concentrations could cause the early death of H. anomala,while the dead S. cerevisiae cells had no inhibition effect on H. anomala. Therefore,the early death of H. anomala was due either to the acids produced by S. cerevisiae and nutrition competition during wine fermentation,or to growth-inhibitory compounds produced by S. cerevisiae and viable S. cerevisiae cells at high concentration. These results were very helpful for thorough understanding of why H. anomala died before S. cerevisiae during wine fermentation.
In order to reveal the characteristics and mechanism of early death of Hansenula anomala,this paper established a mixed fermentation system for Saccharomyces cerevisiae and H. anomala; and studied the effects of acids produced by S. cerevisiae,aerobic fermentation,mixed-culture in fresh medium without oxygen,mixed-culture supernatant,viable or dead S. cerevisiae cells on the growth of H. anomala. The results showed that the growth of H.anomala was inhibited by acid produced by yeast. In the fermentation treatments of aerobic mixed bacteria and anaerobic add fresh medium,the viable counts of H. anomala were all higher than that of anaerobic mixture fermentation; while the viable count of H. anomala in mixed-culture supernatant was lower after adding fresh medium.The presence of viable S. cerevisiae cells at high concentrations could cause the early death of H. anomala,while the dead S. cerevisiae cells had no inhibition effect on H. anomala. Therefore,the early death of H. anomala was due either to the acids produced by S. cerevisiae and nutrition competition during wine fermentation,or to growth-inhibitory compounds produced by S. cerevisiae and viable S. cerevisiae cells at high concentration. These results were very helpful for thorough understanding of why H. anomala died before S. cerevisiae during wine fermentation.
引文
[1] Sadoudi M,Tourdot-Maréchal R,Rousseaux S,et al.. Yeast-yeast interactions revealed by aromatic profile analysis ofSauvignon Blanc wine fermented by single or co-culture of non-Saccharomyces and Saccharomyces yeasts[J]. Food Microbiol.,2012,32(2):243-253.
[2] Gobbi M, Comitini F, Domizio P, et al.. Lachanceathermotolerans and Saccharomyces cerevisiae in simultaneous andsequential co-fermentation:A strategy to enhance acidity andimprove the overall quality of wine[J]. Food Microbiol.,2013,33(2):271-281.
[3] Contreras A,Curtin C,Varela C. Yeast population dynamicsreveal a potential ‘collaboration’ between Metschnikowiapulcherrima and Saccharomyces uvarum for the production ofreduced alcohol wines during Shiraz fermentation[J]. Appl.Microb. Biotechnol.,2015,99(4):1885-1895.
[4] Lee P R,Hui S,Kho C,et al.. Yeast ratio is a critical factorfor sequential fermentation of papaya wine by Williopsis saturnusand Saccharomyces cerevisiae[J]. Microb. Biotechnol.,2012,6(4):385-393.
[5] Ciani M,Beco L,Comitini F. Fermentation behaviour andmetabolic interactions of multi-starter wine yeast fermentations[J]. Int. J. Food Microbiol.,2006,108(2):239-245.
[6] Contreras A,Hidalgo C,Schmidt S,et al.. The application ofnon-Saccharomyces yeast in fermentations with limited aerationas a strategy for the production of wine with reduced alcoholcontent[J]. Int. J. Food Microbiol.,2015,205:7-15.
[7] AndorràI,Berradre M,Mas A,et al.. Effect of mixed culturefermentations on yeast populations and aroma profile[J]. LWTFood Sci. Technol.,2012,49(1):8-13.
[8] Branco P,Francisco D,Chambonc,et al.. Identification ofnovel GAPDH-derived antimicrobial peptides secreted bySaccharomyces cerevisiae and involved in wine microbialinteractions[J]. Appl. Microbiol. Biotechnol.,2014,98(2):843-853.
[9] Taillandier P,Lai Q P,Julien-ortiz A,et al.. Interactionsbetween Torulaspora delbrueckii and Saccharomyces cerevisiae inwine fermentation:Influence of inoculation and nitrogen content[J]. World J. Microb. Biotechnol.,2014,30(7):1959-1967.
[10] Wang C, Mas A, Esteve-zarzoso B. Interaction betweenHanseniaspora uvarum and Saccharomyces cerevisiae duringalcoholic fermentation[J]. Int. J. Food Microbiol.,2015,206:67-74.
[11]中国国家标准化管理委员会. GB/T 4789.2-2010食品卫生微生物学检验菌落总数测定标准[S].北京:中国标准出版社,2010.Standardization Administration of The People's Republic ofChina. GB/T 4789.2-2010 Microbiological examination of foodhygiene———Detection of aerobic bacterial count[S]. Beijing:China Standard Press,2010.
[12] Hanl L,Sommer P,Arneborg N. The effect of decreasingoxygen feed rates on growth and metabolism of Torulasporadelbrueckii[J]. Appl. Microbiol. Biot.,2005,67(1):113-118.
[13] Hansen E H,Nissen P, Sommer P, et al.. The effect ofoxygen on the survival of non-Saccharomyces yeasts duringmixed culture fermentations of grape juice with Saccharomycescerevisiae[J]. J. Appl. Microbiol.,2001,91(3):541-547.
[14] Nissen P,Nielsen D,Arneborg N. The relative glucose uptakeabilities of non-Saccharomyces yeasts play a role in theircoexistence with Saccharomyces cerevisiae in mixed cultures[J]. Appl. Microbiol. Biot.,2004,64(4):543-550.
[15] Kemsawasd V,Viana T,ArdY,et al.. Influence of nitrogensources on growth and fermentation performance of differentwine yeast species during alcoholic fermentation[J]. Appl.Microbiol. Biotechnol.,2015,99(23):10191-10207.
[16] Pérez-nevado F,Albergaria H,Hogg T,et al.. Cellular deathof two non-Saccharomyces wine-related yeasts during mixedfermentations with Saccharomyces cerevisiae[J]. Int. J. FoodMicrobiol.,2006,108(3):336-345.
[17] Albergaria H,Francisco D,Gori K,et al.. Saccharomycescerevisiae CCMI 885 secretes peptides that inhibit the growth ofsome non-Saccharomyces wine-related strains[J]. Appl.Microbiol. Biotechnol.,2010,86(3):965-972.
[18] Albergaria H,Branco P,Francisco D,et al.. Dominance ofSaccharomyces cerevisiae in wine fermentations:Secretion ofantimicrobial peptides and microbial interactions[A]. In:Proceedings International Conference on Microbial Diversity:Microbial Interactions in Complex Ecosystems[C]. Torino,Italy,2013:98-101.
[19] Branco P, Viana T, Albergaria H, et al.. Antimicrobialpeptides(AMPs)produced by Saccharomyces cerevisiae inducealterations in the intracellular pH,membrane permeability andculturability of Hanseniaspora guilliermondii cells[J]. Int. J.Food Microbiol.,2015,205:112-118.
[20] Nissen P,Arneborg N. Characterization of early deaths of non-Saccharomyces yeasts in mixed cultures with Saccharomycescerevisiae[J]. Arch. Microbiol.,2003,180(4):257-263.
[21] Hirano M,Hirano K,Nishimura J,et al.. Transcriptional up-regulation of p27Kip1during contact-induced growth arrest invascular endothelial cells[J]. Exp. Cell Res.,2001,271:356-367.
[22] Kleerebezem M,Quadri L E,Kuipers O P,et al.. Quorumsensing by peptide pheromones and two-component signal-transduction systems in Gram-positive bacteria[J]. Mol.Microbiol.,1997,24(5):895-904.
[23] Liu Y, Rousseaux S, Tourdot-Maréchal R, et al.. Winemicrobiome:A dynamic world of microbial interactions[J].Crit. Rev. Food Sci. Nutr.,2015,57(4):856-873.
[24] Ciani M,Comitini F. Yeast interactions in multi-starter winefermentation[J]. Curr. Opin. Food Sci.,2015,1(1):1-6.
[25] Renault P E,Albertin W,Bely M. An innovative tool revealsinteraction mechanisms among yeast populations underoenological conditions[J]. Appl. Microbiol. Biot.,2013,97(9):4105-4119.
[26] Nehme N,Mathieu F,Taillandier P. Impact of the co-cultureof Saccharomyces cerevisiae-Oenococcus oeni on malolacticfermentation and partial characterization of a yeast-derivedinhibitory peptidic fraction[J]. Food Microbiol.,2010,27(1):150-157.
[2] Gobbi M, Comitini F, Domizio P, et al.. Lachanceathermotolerans and Saccharomyces cerevisiae in simultaneous andsequential co-fermentation:A strategy to enhance acidity andimprove the overall quality of wine[J]. Food Microbiol.,2013,33(2):271-281.
[3] Contreras A,Curtin C,Varela C. Yeast population dynamicsreveal a potential ‘collaboration’ between Metschnikowiapulcherrima and Saccharomyces uvarum for the production ofreduced alcohol wines during Shiraz fermentation[J]. Appl.Microb. Biotechnol.,2015,99(4):1885-1895.
[4] Lee P R,Hui S,Kho C,et al.. Yeast ratio is a critical factorfor sequential fermentation of papaya wine by Williopsis saturnusand Saccharomyces cerevisiae[J]. Microb. Biotechnol.,2012,6(4):385-393.
[5] Ciani M,Beco L,Comitini F. Fermentation behaviour andmetabolic interactions of multi-starter wine yeast fermentations[J]. Int. J. Food Microbiol.,2006,108(2):239-245.
[6] Contreras A,Hidalgo C,Schmidt S,et al.. The application ofnon-Saccharomyces yeast in fermentations with limited aerationas a strategy for the production of wine with reduced alcoholcontent[J]. Int. J. Food Microbiol.,2015,205:7-15.
[7] AndorràI,Berradre M,Mas A,et al.. Effect of mixed culturefermentations on yeast populations and aroma profile[J]. LWTFood Sci. Technol.,2012,49(1):8-13.
[8] Branco P,Francisco D,Chambonc,et al.. Identification ofnovel GAPDH-derived antimicrobial peptides secreted bySaccharomyces cerevisiae and involved in wine microbialinteractions[J]. Appl. Microbiol. Biotechnol.,2014,98(2):843-853.
[9] Taillandier P,Lai Q P,Julien-ortiz A,et al.. Interactionsbetween Torulaspora delbrueckii and Saccharomyces cerevisiae inwine fermentation:Influence of inoculation and nitrogen content[J]. World J. Microb. Biotechnol.,2014,30(7):1959-1967.
[10] Wang C, Mas A, Esteve-zarzoso B. Interaction betweenHanseniaspora uvarum and Saccharomyces cerevisiae duringalcoholic fermentation[J]. Int. J. Food Microbiol.,2015,206:67-74.
[11]中国国家标准化管理委员会. GB/T 4789.2-2010食品卫生微生物学检验菌落总数测定标准[S].北京:中国标准出版社,2010.Standardization Administration of The People's Republic ofChina. GB/T 4789.2-2010 Microbiological examination of foodhygiene———Detection of aerobic bacterial count[S]. Beijing:China Standard Press,2010.
[12] Hanl L,Sommer P,Arneborg N. The effect of decreasingoxygen feed rates on growth and metabolism of Torulasporadelbrueckii[J]. Appl. Microbiol. Biot.,2005,67(1):113-118.
[13] Hansen E H,Nissen P, Sommer P, et al.. The effect ofoxygen on the survival of non-Saccharomyces yeasts duringmixed culture fermentations of grape juice with Saccharomycescerevisiae[J]. J. Appl. Microbiol.,2001,91(3):541-547.
[14] Nissen P,Nielsen D,Arneborg N. The relative glucose uptakeabilities of non-Saccharomyces yeasts play a role in theircoexistence with Saccharomyces cerevisiae in mixed cultures[J]. Appl. Microbiol. Biot.,2004,64(4):543-550.
[15] Kemsawasd V,Viana T,ArdY,et al.. Influence of nitrogensources on growth and fermentation performance of differentwine yeast species during alcoholic fermentation[J]. Appl.Microbiol. Biotechnol.,2015,99(23):10191-10207.
[16] Pérez-nevado F,Albergaria H,Hogg T,et al.. Cellular deathof two non-Saccharomyces wine-related yeasts during mixedfermentations with Saccharomyces cerevisiae[J]. Int. J. FoodMicrobiol.,2006,108(3):336-345.
[17] Albergaria H,Francisco D,Gori K,et al.. Saccharomycescerevisiae CCMI 885 secretes peptides that inhibit the growth ofsome non-Saccharomyces wine-related strains[J]. Appl.Microbiol. Biotechnol.,2010,86(3):965-972.
[18] Albergaria H,Branco P,Francisco D,et al.. Dominance ofSaccharomyces cerevisiae in wine fermentations:Secretion ofantimicrobial peptides and microbial interactions[A]. In:Proceedings International Conference on Microbial Diversity:Microbial Interactions in Complex Ecosystems[C]. Torino,Italy,2013:98-101.
[19] Branco P, Viana T, Albergaria H, et al.. Antimicrobialpeptides(AMPs)produced by Saccharomyces cerevisiae inducealterations in the intracellular pH,membrane permeability andculturability of Hanseniaspora guilliermondii cells[J]. Int. J.Food Microbiol.,2015,205:112-118.
[20] Nissen P,Arneborg N. Characterization of early deaths of non-Saccharomyces yeasts in mixed cultures with Saccharomycescerevisiae[J]. Arch. Microbiol.,2003,180(4):257-263.
[21] Hirano M,Hirano K,Nishimura J,et al.. Transcriptional up-regulation of p27Kip1during contact-induced growth arrest invascular endothelial cells[J]. Exp. Cell Res.,2001,271:356-367.
[22] Kleerebezem M,Quadri L E,Kuipers O P,et al.. Quorumsensing by peptide pheromones and two-component signal-transduction systems in Gram-positive bacteria[J]. Mol.Microbiol.,1997,24(5):895-904.
[23] Liu Y, Rousseaux S, Tourdot-Maréchal R, et al.. Winemicrobiome:A dynamic world of microbial interactions[J].Crit. Rev. Food Sci. Nutr.,2015,57(4):856-873.
[24] Ciani M,Comitini F. Yeast interactions in multi-starter winefermentation[J]. Curr. Opin. Food Sci.,2015,1(1):1-6.
[25] Renault P E,Albertin W,Bely M. An innovative tool revealsinteraction mechanisms among yeast populations underoenological conditions[J]. Appl. Microbiol. Biot.,2013,97(9):4105-4119.
[26] Nehme N,Mathieu F,Taillandier P. Impact of the co-cultureof Saccharomyces cerevisiae-Oenococcus oeni on malolacticfermentation and partial characterization of a yeast-derivedinhibitory peptidic fraction[J]. Food Microbiol.,2010,27(1):150-157.