Overview of a Surface-Ripened Cheese Community Functioning by Meta-Omics Analyses
Éric Dugat‐BonyCécile StraubAurélie TeissandierDjamila OnésimeValentin LouxChristophe MonnetFrançoise IrlingerSophie LandaudM.-N. Leclercq-PerlatPascal BentoSébastien FraudJean‐François GibratJulie AubertF. FérÉric GuédonNicolas PonsSean KennedyJean-Marie BeckerichDominique SwennenPascal Bonnarme
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Abstract:
Cheese ripening is a complex biochemical process driven by microbial communities composed of both eukaryotes and prokaryotes. Surface-ripened cheeses are widely consumed all over the world and are appreciated for their characteristic flavor. Microbial community composition has been studied for a long time on surface-ripened cheeses, but only limited knowledge has been acquired about its in situ metabolic activities. We applied metagenomic, metatranscriptomic and biochemical analyses to an experimental surface-ripened cheese composed of nine microbial species during four weeks of ripening. By combining all of the data, we were able to obtain an overview of the cheese maturation process and to better understand the metabolic activities of the different community members and their possible interactions. Furthermore, differential expression analysis was used to select a set of biomarker genes, providing a valuable tool that can be used to monitor the cheese-making process.Keywords:
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It takes long time for cheese ripening,which lead to the increase of the manufacture cost.Accelerated ripening of cheese is one of the effective ways to reduce it.Approaches have been used frequently in cheese ripening include the addition of enzymes,attenuated starter cells,elevated ripening temperatures,high pressure treatment and so on,however there are some defects for each other.Non-starter lactic acid bacteria(NSLAB) could accelerate cheese ripening,so it become one of focus of the methods to accelerate cheese ripening.The roles of microorganisms on the cheesemaking and ripening,the reason for using adjunct culture and its advances will be surveyed in this paper.
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Four cheesemakings of Graviera Kritis cheese (chee ses A-D) were made from a mixture of ewe' s and goal' s milk.Three of these were made with mixed cultures of thermophilic + propionic or mesophilic + thermophilic + propionic starters, while the fourth one was made traditionally without starter cultures.The effect of increasing the temperature of ripening was also determined.The pH 4.4-soluble nitrogen (SN) and trichloroacetic acid (TCA-SN) fractions increased mainly during the warm room ripening.The use of starters caused no significant differences in the ratios of nitrogen fractions of the four mature cheeses (age> 90 d), whatever the ripening temperature.Nitrogen fractions were significantly correlated with cheese age.At 180 d of ripening, the percentage of pH 4.4-SN in total nitrogen (TN) was 20.7-22.8 % and that of TCA-SN in TN was 19.1-19.3%.Polyacrylamide gel electrophoretic (PAGE) results showed that chymosin and plasmin action was intense during the maturation of the cheeses.The proportion of hydrophilic peptides was higher in the water soluble nitrogen (WSN) fraction of the cheeses made with starter cultures at both ripening temperatures.Free amino acids (FAA) were not significantly different in the four types of cheeses, reaching 208-246 mmol•kg-' after 180 d of ripening.The changes in total FAA and in the most abundant FAA during the ripening were significantly correlated with chee se age (r > 0.940).The ripening at higher temperature increased the main FAA and the total FAA content in ail types of cheeses, in particular in the chee se made without starter cultures.© InralElsevier, Paris.
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To accelerate cheese ripening, enhance its flavor types and intensity and make cheese flavoring agent in shorter time, surface ripening bacterium (Brevibacterium linens and Debaryomyces hansenii) and/or enzymes (Flavorzyme 500 MG and Palatase 20000 L) were used in cheese curd. In this study, aroma compounds generated by using ripening cultures and/or enzymes were analyzed. The control l was made by inoculating ripening cultures, while the control 2 was made through enzymes-modified only. Results showed that cheese flavoring agent made by using ripening strains in combination with enzymes had more volatile flavor compounds (at least 44) than that used just ripening bacterium (26) or just two enzymes (27). Then, through Solid-phase microextraction and Gas Chromatography-Mass Spectrometry analysis, we knew that sample 1, which was made through proteolysis first, next sprayed ripening cultures and last lipolysis, generated 54 flavor compounds. Sample 2, which enzymed cheese curd first, then incubated ripening cultures, had 44 aroma compounds. However, the controls 1, incubated ripening strains only, had 26 volatile compounds, while the control 2, enzymed only, had 27 volatile compounds. This study reveals that ripening bacterium could contribute more to the generation of acids, sulphur compounds, miscellaneous compounds and alcohols, it has a good potential to be used in cheese flavoring agents making. Besides, the combination of surface strains and enzymes, especially using Flavorzyme 500 MG first, then sprayed ripening cultures and at last Palatase 20000 L, could get more volatile compounds.
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Monitoring Proteolysis During Ripening of Full‐fat and Low‐fat Cheddar Cheeses by Reverse‐Phase HPLC
ABSTRACT Proteolysis during ripening of full‐fat and low‐fat Cheddar cheese was investigated by applying reverse‐phase HPLC to the pH 4.6 water‐soluble N fraction of cheese. The separated N compounds were divided into four MW ranges. The number of separated peaks and the amount of N compounds separated in each of the MW ranges increased with ripening time. Significant within‐cheese, within‐variety, and between‐varieties differences in the amounts and proportions of N compounds with different MW were observed as related to ripening time. Reverse‐phase HPLC of the pH 4.6 water‐soluble N fraction of cheese can provide detailed quantitative information on proteolytic activities during ripening.
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This experiment was carried out to study the ripening of control cheese made with lactic acid bacteria only and yeast cheese made with lactic acid bacteria and yeast (C. lipolitica, C. pseudotropicalis, S. fragilis, D. hansenii 4193), with 0, 2, and 5% levels of salt content by the modified kosikowski`s method (1977). The work reported here determined the degree of proteolysis in the cheese for 12 weeks of ripening under 5, 10℃ ripening temperature. The results obtained are summarized as followed: 1. WSN and soluble tyrosine contents in cheese were increased with ripening period. WSN contents of yeast cheese were higher significantly (p$lt;0.05) than those of control cheese at 12 weeks of ripening. Also WSN and soluble tyrosine contents of unsalted cheeses and the cheeses ripened at 10℃ generally had higher than those of other cheeses. Average WSN contents of control cheese (No. 1) and yeast cheese (No. 2,3,4,S, 6) increased from 0.31, 0.33, 0.58, 0.30, 0.34, and 0.30%, respectively after manufacture to 1.24, 1.45, 1.53, 1.49, 1.61, and 1.45%, respectively at 12 weeks of ripening. The ripening period, salt contents and ripening temperature had significant effects (p$lt;0.01) on the WSN and soluble tyrosine contents.
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