不同发酵条件对模拟葡萄酒中酒酒球菌柠檬酸代谢的影响
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摘要
以酒酒球菌31-DH进行模拟酒苹果酸-乳酸发酵,使用离子排斥色谱法测定相关代谢物质的含量,研究苹果酸-乳酸发酵期间及发酵后p H、乙醇和葡萄糖对柠檬酸代谢的影响。结果表明:该方法耗时短、前处理简单、分离度良好、回收率为87%~108%、精密度为0.77%~4.33%,满足分析要求。当发酵液处于低p H,高酒精度,低葡萄糖浓度时,柠檬酸代谢缓慢,乳酸、乙酸及2,3-丁二醇的产量较低,双乙酰含量峰值较高。当发酵液中不含有乙醇时,苹果酸、柠檬酸、葡萄糖代谢迅速,伴随菌体的大量生长,乳酸、乙酸、2,3-丁二醇含量较高,但无双乙酰的产生。当发酵液中不含有葡萄糖时,柠檬酸代谢速率加快,乳酸产量较低,2,3-丁二醇及双乙酰产量较高。p H、乙醇、葡萄糖均显著影响酒酒球菌31-DH对柠檬酸的代谢。
With Oenococcus oeni 31- DH as material for malolactic fermentation,the effects of p H,ethanol and glucose on the citrate metabolism during and after malolactic fermentation were studied.Ion exclusion chromatography was used to measure the contents of the related substances. The results showed that the experiment was time- saving with simple pretreatments,good separation degree,recovery rate between 87% ~ 108% and RSD between 0.77% ~ 4.33% which met the requirements of analysis.With lower p H values,higher ethanol contents and lower glucose contents of the fermentation broth,the citrate metabolism was slower,the lactic acid,acetic acid and 2,3- butanediol contents were lower,while the diacetyl content peak was higher.When the fermentation broth contained no ethanol,the malic acid,citric acid and glucose were metabolized rapidly,associated with mass growth of bacteria,higher contents of lactic acid,acetic acid and 2,3- butanediol,while no diacetyl.When the fermentation broth contained no glucose,the rate of citrate metabolism accelerated,lactic acid production was low and the production of 2,3- butanediol and diacetyl was high.p H,ethanol and glucose had significant effects on the citrate metabolism of O.oeni.
With Oenococcus oeni 31- DH as material for malolactic fermentation,the effects of p H,ethanol and glucose on the citrate metabolism during and after malolactic fermentation were studied.Ion exclusion chromatography was used to measure the contents of the related substances. The results showed that the experiment was time- saving with simple pretreatments,good separation degree,recovery rate between 87% ~ 108% and RSD between 0.77% ~ 4.33% which met the requirements of analysis.With lower p H values,higher ethanol contents and lower glucose contents of the fermentation broth,the citrate metabolism was slower,the lactic acid,acetic acid and 2,3- butanediol contents were lower,while the diacetyl content peak was higher.When the fermentation broth contained no ethanol,the malic acid,citric acid and glucose were metabolized rapidly,associated with mass growth of bacteria,higher contents of lactic acid,acetic acid and 2,3- butanediol,while no diacetyl.When the fermentation broth contained no glucose,the rate of citrate metabolism accelerated,lactic acid production was low and the production of 2,3- butanediol and diacetyl was high.p H,ethanol and glucose had significant effects on the citrate metabolism of O.oeni.
引文
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[15]Da Silverira M G,Abee T.Activity of ethanol-stresssed Oenococcus oeni cells:a flow cytometric approach[J].Journal of Applied Microbiology,2009,106:1690-1696.
[16]Lonvaud-Funel A,Zmirou C,Larue F.Le métabolisme de l’acide citrique par les bactéries lactiques de la fermentation malolactique des vins[J].Sci Aliments,1984,4:81-85.
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[2]李瑞国,韩烨,周志江.葡萄酒苹果酸乳酸发酵研究进展[J].食品研究与开发,2010,31(8):228-233.
[3]Ruiz P,Izquierdo P M,Sesena S,et al.Malolactic Fermentation and Secondar Metabolite Production by Oenoccocus oeni Strains in Low p H Wines[J].Journal of Food Science,2012,77(10):579-585.
[4]Lonvaud-Funel A.Lactic acid bacteria in the quality improvement and depreciation of wine[J].Antonie van Leewenhoek,1999,76:317-331.
[5]Latitia G,Pascal D,Yann D.The Citrate Metabolism in Homo-and Heterofermentative LAB:A Selective Means of Becoming Dominant over Other Microorganisms in Complex Ecosystems[J].Food and Nutrition Sciences,2014,5:953-969.
[6]Quintáns G N,Repizo G,Magni C,et al.Citrate Metabolism and Aroma Compound Production in Lactic acid Bacteria[C].In:Mayo B,López P,Pérez-Martínez G,Eds.,Molecular Aspects of Lactic acid Bacteria for Traditional and New Applications.Research Signpost,Kerala,India,2008:65-88.
[7]Bartowsky E J,Henschke P A.The‘buttery’attribute of wine-diacetyl-desirability,spoilage and beyond[J].International Journal of Food Microbiology,2004,96(3):235-252.
[8]林琼.柠檬酸代谢及转运相关基因对柑橘果实酸度的调控机制[D].杭州:浙江大学,2015.
[9]张规富,谢深喜.柑橘果实柠檬酸代谢研究进展[J].中国园艺文摘,2012(8):1-4.
[10]韦函忠,马少敏,韦朝英.啤酒发酵中双乙酰的产生和α-乙酰乳酸脱羧酶的应用[J].广西轻工业,2011(8):17-18,34.
[11]李亚辉.酒类酒球菌SD-2a和31MBR的β-D-葡萄糖苷酶研究[D].杨凌:西北农林科技大学,2013.
[12]Krieger S A,Lempere E,Ernst M.Management of malolactic fermentation with regard to flavour modification.Session 3B,Flavour modification in the winery:microbiological[C].Proceedings of 5th International Symposium on Cool Climate Viticulture and Oenology.Melbourne,Australia,2000:1-4.
[13]García-Quintáns N,Repizo G,Magni M,et al.Activation of the Diacetyl/Acetoin Pathway in Lactococcus lactis subsp.lactis bv.diacetylactis CRL264 by Acidic Growth[J].Applied and Enviromental Microbiology,2008,74(7):1988-1996.
[14]Olguin N,Bordons A,Reguant C.Influence of ethanol and p H on the gene expression of the citrate pathway in Oenococcus oeni[J].Food Microbiology,2009,26:197-203.
[15]Da Silverira M G,Abee T.Activity of ethanol-stresssed Oenococcus oeni cells:a flow cytometric approach[J].Journal of Applied Microbiology,2009,106:1690-1696.
[16]Lonvaud-Funel A,Zmirou C,Larue F.Le métabolisme de l’acide citrique par les bactéries lactiques de la fermentation malolactique des vins[J].Sci Aliments,1984,4:81-85.
[17]Salou P,Loubiere P,Pareilleux A.Growth and energetics of Leconostoc oenos during comatabolism of glucose with citrate or gructose[J].Applied and Enviromental Microbiology,1994,60(5):1459-1466.