Reverse transcription quantitative PCR revealed persistency of thermophilic lactic acid bacteria metabolic activity until the end of the ripening of Emmental cheese

Helene Falentin; Nadine Henaff; Pierre Le Bivic; Stephanie-Marie Deutsch; Sandrine Parayre; Romain Richoux; Daniele Sohier; Anne Thierry; Sylvie Lortal; Florence Postollec

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Reverse transcription quantitative PCR revealed persistency of thermophilic lactic acid bacteria metabolic activity until the end of the ripening of Emmental cheese

机译：逆转录定量PCR显示嗜热乳酸菌的代谢活性持续存在，直至埃门塔尔干酪成熟

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For Emmental manufacture two kinds of adjunct culture are added: (i) thermophilic lactic acid bacteria (starters) such as Lactobacillus helveticus (LH), and Streptococcus thermophilus (ST) growing the first day of the manufacture and (ii) ripening culture. STand LH have a key role in curd acidification and proteolysis at the beginning of the manufacture but are considered to be lyzed for a great part of them at the ripening step. The aim of this work was to assess the metabolic activity of these bacteria throughout manufacture and ripening. During Emmental cheesemaking, LH and ST were subjected to i) population quantification by numerations and by quantitative PCR (qPCR) ii) reverse transcription (RT) Temporal Temperature Gel Electrophoresis (TTGE) iii) transcript quantification by RT-qPCR targeting 16S rRNA, tuf and grol mRNAs to evaluate bacterial metabolic activity. During ripening, STand LH numerations showed a 2.5 logio loss of culturability whereas qPCR on pelleted cells revealed only one logio of decrease for both of these species. 109 STand 108 LH cells/g of cheese still remained. They contained a stable number of 16S transcript and at least 106 copies of mRNAs per 109 cells until the end of ripening. These results prove the unexpected persistency .of thermophilic lactic acid bacteria starters (ST and LH) metabolic activity until the end of ripening and open new perspectives in term of their involvement in the quality of cheeses during ripening.

机译：对于Emmental制造，添加了两种辅助培养：（i）嗜热乳酸菌（发酵剂），如瑞士乳杆菌（LHtobacillus helveticus，LH），以及在生产的第一天生长的嗜热链球菌（ST），以及（ii）培养成熟。 STand LH在生产开始时就对凝乳酸化和蛋白水解起关键作用，但在成熟步骤中，大部分被认为可将其溶解。这项工作的目的是评估这些细菌在整个制造和成熟过程中的代谢活性。在Emmental奶酪制作过程中，对LH和ST进行i）通过计数和定量PCR（qPCR）进行种群定量ii）反转录（RT）时温凝胶电泳（TTGE）iii）通过针对16S rRNA，tuf的RT-qPCR进行转录本定量和grol mRNAs来评估细菌的代谢活性。在成熟过程中，STand LH计数显示可培养性下降了2.5 log10，而沉淀细胞上的qPCR显示这两个物种仅下降了1 logio。仍保留109 STand 108 LH细胞/克奶酪。它们包含稳定数量的16S转录物和每109个细胞至少106个拷贝的mRNA，直至成熟。这些结果证明了嗜热乳酸菌发酵剂（ST和LH）的代谢活动具有出乎意料的持久性，直到成熟结束为止，并为它们在成熟过程中对奶酪品质的影响开辟了新的视野。

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来源
《Food microbiology》 |2012年第1期|p.132-140|共9页
作者
Helene Falentin; Nadine Henaff; Pierre Le Bivic; Stephanie-Marie Deutsch; Sandrine Parayre; Romain Richoux; Daniele Sohier; Anne Thierry; Sylvie Lortal; Florence Postollec;
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作者单位

INRA, UMR 1253, Science et Technologie du Lait et de Voeuf, F-35000, Rennes, France,AGROCAMPUS OUEST, UMR12S3, Science et Technologie du Lait et de Voeuf, F-35000 Rennes, France;

ADRIA Developpement, F-29000, Quimper, France;

INRA, UMR 1253, Science et Technologie du Lait et de Voeuf, F-35000, Rennes, France,AGROCAMPUS OUEST, UMR12S3, Science et Technologie du Lait et de Voeuf, F-35000 Rennes, France;

INRA, UMR 1253, Science et Technologie du Lait et de Voeuf, F-35000, Rennes, France,AGROCAMPUS OUEST, UMR12S3, Science et Technologie du Lait et de Voeuf, F-35000 Rennes, France;

INRA, UMR 1253, Science et Technologie du Lait et de Voeuf, F-35000, Rennes, France,AGROCAMPUS OUEST, UMR12S3, Science et Technologie du Lait et de Voeuf, F-35000 Rennes, France;

AOILAIT, F-35000, Rennes, France;

ADRIA Developpement, F-29000, Quimper, France;

INRA, UMR 1253, Science et Technologie du Lait et de Voeuf, F-35000, Rennes, France,AGROCAMPUS OUEST, UMR12S3, Science et Technologie du Lait et de Voeuf, F-35000 Rennes, France;

INRA, UMR 1253, Science et Technologie du Lait et de Voeuf, F-35000, Rennes, France,AGROCAMPUS OUEST, UMR12S3, Science et Technologie du Lait et de Voeuf, F-35000 Rennes, France;

ADRIA Developpement, F-29000, Quimper, France;

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收录信息美国《科学引文索引》(SCI);美国《化学文摘》(CA);
原文格式 PDF
正文语种 eng
中图分类
关键词
swiss cheese; ripening; RT-Qpcr; gene expression; lactic acid bacteria; starter;

机译：瑞士芝士;成熟RT-Qpcr;基因表达;乳酸菌起动机;

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1. Specific metabolic activity of ripening bacteria quantified by real-time reverse transcription PCR throughout Emmental cheese manufacture. [J] . Falentin H, Postollec F, Parayre S, International Journal of Food Microbiology . 2010,第1期

机译：在整个埃门塔尔干酪生产中，通过实时逆转录PCR定量的成熟细菌的比代谢活性。
2. Effect of cook temperature on starter and non-starter lactic acid bacteria viability, cheese composition and ripening indices of a semi-hard cheese manufactured using thermophilic cultures. [J] . Sheehan J J, Fenelon M A, Wilkinson M G, International Dairy Journal . 2007,第6期

机译：蒸煮温度对使用嗜热培养物生产的半硬干酪的起子和非起子乳酸菌活力，干酪组成和成熟指数的影响。
3. Investigation of Geotrichum candidum gene expression during the ripening of Reblochon-type cheese by reverse transcription-quantitative PCR [J] . Castellote Jessie, Fraud Sebastien, Irlinger Francoise, International Journal of Food Microbiology . 2015,第Null期

机译：通过逆转录定量PCR研究再生型奶酪成熟过程中Geotrichum Candidum基因表达的研究
4. EFFECT OF THE TYPE OF STARTER CULTURE ADDED TO THE MILK POST-THERMIZATION ON THE COMPOSITION OF THE LACTIC ACID BACTERIA FLORA DOMINATING IN TRADITIONAL FULLY RIPENED GRAVIERA CHEESE [C] . J. Samelis, A. Kakouri Πανελληνιο Συνεδριο ΤροΦιμων . 2012

机译：高温后牛奶发酵中添加的发酵剂类型对乳酸菌在完全全缘干酪干酪中的组成的影响
5. Evaluation of lactic acid bacteria for the acceleration of cheese ripening using pulsed electric fields. [D] . Briggs, Stephanie Sheryl. 2003

机译：使用脉冲电场评估乳酸菌对奶酪成熟的促进作用。
6. The hsp 16 Gene of the Probiotic Lactobacillus acidophilus Is Differently Regulated by Salt High Temperature and Acidic Stresses as Revealed by Reverse Transcription Quantitative PCR (qRT-PCR) Analysis [O] . Vittorio Capozzi, Mattia Pia Arena, Elisabetta Crisetti, 2011

机译：逆转录定量PCR（qRT-PCR）分析表明盐高温和酸性胁迫对益生菌嗜酸乳杆菌的hsp 16基因有不同的调控作用。
7. The hsp 16 Gene of the Probiotic Lactobacillus acidophilus Is Differently Regulated by Salt, High Temperature and Acidic Stresses, as Revealed by Reverse Transcription Quantitative PCR (qRT-PCR) Analysis [O] . Capozzi, Vittorio, Arena, Mattia Pia, Crisetti, Elisabetta, 2011

机译：逆转录定量PCR（qRT-PCR）分析显示，盐，高温和酸性胁迫对益生菌嗜酸乳杆菌的hsp 16基因有不同的调控作用。

Reverse transcription quantitative PCR revealed persistency of thermophilic lactic acid bacteria metabolic activity until the end of the ripening of Emmental cheese

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