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Isovaleric Acid Is Mainly Produced by Propionibacterium Freudenreichii in Swiss Cheese Anne Thierry, Romain Richoux, Jean-René Kerjean

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Isovaleric Acid Is Mainly Produced by Propionibacterium Freudenreichii in Swiss Cheese Anne Thierry, Romain Richoux, Jean-René Kerjean Isovaleric acid is mainly produced by Propionibacterium freudenreichii in Swiss cheese Anne Thierry, Romain Richoux, Jean-René Kerjean To cite this version: Anne Thierry, Romain Richoux, Jean-René Kerjean. Isovaleric acid is mainly produced by Pro- pionibacterium freudenreichii in Swiss cheese. International Dairy Journal, Elsevier, 2004, 14 (9), pp.801-807. 10.1016/j.idairyj.2004.02.002. hal-02264923 HAL Id: hal-02264923 https://hal.archives-ouvertes.fr/hal-02264923 Submitted on 31 May 2020 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. ARTICLE IN PRESS International Dairy Journal 14 (2004) 801–807 Isovaleric acid is mainly produced by Propionibacterium freudenreichii in Swiss cheese Anne Thierrya,*, Romain Richouxb, Jean-Rene! Kerjeanb a Laboratoire de Recherches de Technologie Laitiere," INRA., 65 rue de Saint-Brieuc, Rennes Cedex 35042, France b Institut Technique Fran@ais des Fromages, BP 6224, Rennes Cedex 35062, France Received 7 July 2003; accepted 18 February 2004 Abstract Isovaleric acid (3-methylbutyric acid) and 2-methylbutyric acid contribute to Swiss cheese flavour. In order to determine the contribution of propionibacteria (PAB) to the production of methylbutyric acids, mini-Swiss cheeses were manufactured with or without PAB as a secondary starter (25 Propionibacterium freudenreichii strains), associated with different cultures of thermophilic lactic starters. In the presence of PAB, the quantity of methylbutyric acids was three to ten times greater, depending on PAB strain, than in the absence of PAB, regardless of the species and the strain of lactobacilli used (20–63 vs. o6mgkgÀ1 ripened cheese). PAB produced methylbutyric acids concomitantly with acetic and propionic acids, then kept on producing methylbutyric acids after propionic fermentation. The increase in salt-in-moisture content of cheese from 1.0% to 1.8% induced a strain-dependent inhibition of isovaleric acid production. This study shows that P. freudenreichii is the main contributor to methylbutyric acid production in Swiss cheese. r 2004 Elsevier Ltd. All rights reserved. Keywords: Propionibacterium; Isovaleric acid; Swiss-type cheese; Flavour; Thermophilic lactic starter; Salt 1. Introduction Buchin, Hauwuy, & Coulon, 2001). The concentration of isovaleric acid in Swiss cheeses varies over a large Flavour compounds are formed during the cheese range (from undetectable values up to 100 mg kgÀ1 ripening process from the enzymatic breakdown of milk cheese) (Patton, 1964; Langler & Day, 1966; Bosset et al., protein, fat, lactose and lactate (McSweeney, Nursten, & 1992), withmean values of around 26 mg kg À1 in French Urbach, 1997). However, we have only fragmentary Emmental cheese (Berdague,! Grappin, Delacroix- knowledge of the microflora and the pathways involved Buchet, & Chaillet, 1990). In many of the available in the formation of many flavour compounds. To reports, the term ‘‘isovaleric acid’’ referred to the control and diversify cheese flavour, it is necessary to mixture of 3-methyl-butyric acid and 2-methyl-butyric understand how microflora and other factors affect the acid, which were not separated by the chromatographic formation and synthesis pathways for each family of method used to analyse volatile acids. In this paper, the flavour compounds. term ‘‘methylbutyric acids’’ was preferred to refer to the 3-Methylbutyric acid (actual isovaleric acid) and mixture of bothacids. 3-Methylbutyric acid and 2-methylbutyric acid are branched-chain volatile acids 2-methylbutyric acid originate from leucine and iso- involved in Swiss cheese flavour (Langler, Libbey, & leucine catabolism, respectively, via a first step of Day, 1967; Preininger, Warmke, & Grosch, 1996). The transamination followed by oxidation steps. presence of isovaleric acid is related to the intensity and The microflora that produce methylbutyric acids in aromatic richness of several cheeses such as Comte! Swiss cheese have never been completely clarified. Most (Berdague! & Grappin, 1988) and Abondance (Bugaud, species of Swiss cheese ecosystem are able to produce methylbutyric acids in vitro, as recently shown using *Corresponding author. Tel.: +33-2-2348-5337; fax: +32-2-2348- resting cells of different species incubated in the presence 5350. of leucine and a-ketoglutarate: Lactobacillus helveticus, E-mail address: [email protected] (A. Thierry). Lactobacillus delbrueckii subsp. lactis (Yvon & Rijnen, 0958-6946/$ - see front matter r 2004 Elsevier Ltd. All rights reserved. doi:10.1016/j.idairyj.2004.02.002 ARTICLE IN PRESS 802 A. Thierry et al. / International Dairy Journal 14 (2004) 801–807 2001; Helinck, Moreau, & Yvon, 2001), Propionibacter- Kerjean, 1995; Richoux, Faivre, & Kerjean, 1998). The ium freudenreichii (Thierry, Maillard, & Yvon, 2002), starters were composed of S. thermophilus TA060, and Lactobacillus paracasei, a non-starter bacterium L. helveticus and/or L. delbrueckii subsp. lactis, withor (Tammam, Williams, Noble, & Lloyd, 2000; Williams, without the addition of P. freudenreichii as secondary Noble, & Banks, 2001; Tavaria, Dahl, Carballo, & starters. Briefly, the milk was ripened with starters at Malcata, 2002). However, methylbutyric acids appeared 32C for 1 hand coagulated by theaddition of calf only in samples containing P. freudenreichii in cream rennet (Chr Hansen, Beaune, France). When the desired fermented by Swiss cheese starter organisms both singly firmness was reached, the gel was cut into pieces of and in combinations at temperature and pH simulating 4 mm, stirred 15 min and cooked at 53.5C for 45 min. cheese ripening conditions (Biede, Paulsen, Hammond, The pressing step (6 h) and the acidification step (14 h) & Glatz, 1979). Similarly, methylbutyric acid were were conducted in thermostated ovens at 48C and found mainly produced by P. freudenreichii (one strain 36C, respectively, in order to mimic the cooling rate of tested) in small-scale Swiss cheeses (Thierry & Maillard, the centre of a full-size Emmental wheel. Cheeses were 2002). Our hypothesis was that P. freudenreichii is the then cooled and salted in a NaCl-saturated brine at 7C organism responsible for the majority of the methylbu- for 3.5 or 21 h(to obtain, respectively, 1% and 1.8% tyric acid production in Swiss-type cheese. To check this salt-in-moisture). Eachcheesewas manufactured in hypothesis, we determined the production of methylbu- triplicate. Cheeses were divided into four pieces (200 g tyric acids in small-scale Swiss cheeses manufactured each) before waxing. Cheeses were ripened at 11C (cold with or without the addition of P. freudenreichii as room) for 21 days and then at 24C (warm room) for 28 secondary starter (25 strains tested), in the presence of days. One piece was used for all the analyses made at different combinations of thermophilic lactic starters. each ripening stage (cheeses entering in the warm room, We showed that methylbutyric acids were mainly and at different times of ripening in the warm room: d14 produced by P. freudenreichii in Swiss cheese and that and d28 (end) for all cheeses, and two added ripening the production was strain-dependent. stages (d7 and d21) for some cheeses, in order to determine the time course of short-chain carboxylic acid production. 2. Materials and methods The data presented in this manuscript refer to cheeses made using the same standardised protocol from 1991 to 2.1. Starters 1996, for the earlier studies of Richoux and Kerjean (1995), Richoux et al. (1998). These experiments aimed Several thermophilic lactic starters and PAB starters at screening PAB strains for their ability to produce were used. Pure cultures of L. helveticus (ITGLH1, acetate, propionate and opening in cheese (Richoux ITGLH77 and LHRR), L. delbrueckii subsp. lactis et al., 1998). Volatile acids (from acetic to hexanoic (ITGLL14, ITGLL51), S. thermophilus (ITGST49/50) acids) were analysed at that time. Although a peak of and P. freudenreichii (ITGP1, ITGP3, ITGP4, ITGP5, ‘‘isovaleric acid’’ (2-methyl+3-methylbutyric acids) was ITGP6, ITGP8, ITGP9, ITGP10, ITGP11, ITGP12, detected and integrated in the chromatograms of volatile ITGP13, ITGP14, ITGP15, ITGP16, ITGP17, ITGP18, acids, these data were not systematically dealt with at ITGP19, ITGP20, ITGP21, ITGP22, ITGP23, ITGP24, that time. Among these different sets of data, we ITGP25) were obtained from the collection of the selected: (1) all the trials in which different strains of Institut Technique Fran@ais du Fromage (35062 Re- thermophilic lactobacilli were used as starters (with or nnes, France). P. freudenreichii strains A and B are two without added PAB); (2) trials carried out with the same industrial strains from the TL collection (Laboratoire de strains of lactic starters (TA060+LH100) and various Recherches de Technologie Laitiere," INRA, 35042 PAB strains, at two levels of salt-in-moisture (Richoux Rennes, France), and L. helveticus LHRR was an et al., 1998). industrial strain from UCC Culture Collection (Cork, Ireland). LH100 (L. helveticus and L. delbrueckii subsp. 2.3. Biochemical analyses lactis) and S.
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