Abstract
Flor yeasts confer a wide range of organoleptic properties to Sherry-type wines during a process called “biological aging” that takes place after alcoholic fermentation. These kinds of yeasts adapt to a biological aging condition by forming a biofilm known as “flor velum” and by changing from fermentative to oxidative metabolism. It has been reported that some functions such as increase of cell surface hydrophobicity or changes to lipid metabolism are enhanced when yeasts switch to biofilm lifestyle. Here, we attempt to reveal intracellular metabolites and protein molecular functions not documented before that are relevant in biofilm formation and in fermentation by an endometabolome and proteome screening. We report that at early stages of biofilm formation, flor yeasts accumulate mannose, trehalose, glycerol, oleic and stearic acids and synthesize high amounts of GTPases, glycosylases and lipoproteins. On the other hand, in early fermentation, flor yeasts rapidly consume glucose and phosphoric acid; and produce abundant proteins related to chromatin binding, transcription factors and methyl transferases.
Similar content being viewed by others
References
Albertyn J, Hohmann S, Thevelein JM, Prior BA (1994) GPD1, which encodes glycerol-3-phosphate dehydrogenase is essential for growth under osmotic stress in Saccharomyces cerevisiae and its expression is regulated by the high-osmolarity glycerol response pathway. Mol Cell Biol 14:4135–4144. https://doi.org/10.1128/MCB.14.6.4135
Alexandre H, Blanchet S, Charpentier C (2000) Identification of a 49 kDa hydrophobic cell wall mannoprotein present in velum yeast which may be implicated in velum formation. FEMS Microbiol Lett 185:147–150. https://doi.org/10.1111/j.1574-6968.2000.tb09053.x
Ansell R, Granath K, Hohmann S, Thevelein JM, Adler L (1997) The two isoenzymes for yeast NAD+-dependent glycerol 3-phosphate dehydrogenase encoded by GPD1 and GPD2 have distinct roles in osmoadaptation and redox regulation. EMBO J 16:2179–2187. https://doi.org/10.1093/emboj/16.9.2179
Bell W, Klaassenm P, Ohnacker M. Boller T, Herweijer M, Schoppink P, Van der Zee P, Wiemken A (1992) Characterization of the 56-kDa subunit of yeast trehalose-6-phosphate synthase and cloning of its gene reveal its identity with the product of CIF1, a regulator of carbon catabolite inactivation. Eur J Biochem 209:951–959. https://doi.org/10.1111/j.1432-1033.1992.tb17368.x
Blum S, Schmid SR, Pause A, Buser P, Linder P, Sonenberg N, Trachsel H (1992) ATP hydrolysis by initiation factor 4A is required for translation initiation in Saccharomyces cerevisiae. Proc Natl Acad Sci USA 89:7664–7668. https://doi.org/10.1073/pnas.89.16.7664
Carlson M, Botstein D (1982) Two differentially regulated mRNAs with different 5′ ends encode secreted and intracellular forms of yeast invertase. Cell 28:145–154. https://doi.org/10.1016/0092-8674(82)90384-1
Esteve-Zarzoso B, Peris-Torán MJ, García-Maiquez E, Uruburu F, Querol A (2001) Yeast population dynamics during the fermentation and biological aging of Sherry wines. Appl Environ Microbiol 67:2056–2061. https://doi.org/10.1128/AEM.67.5.2056-2061.2001
Farkas I, Hardy TA, Goebl MG, Roach PJ (1991) Two glycogen synthase isoforms in Saccharomyces cerevisiae are coded by distinct genes that are differentially controlled. J Biol Chem 266:15602–15607
Farris GA, Sinigaglia M, Budroni M, Guerzoni ME (1993) Cellular fatty acid composition in film-forming strains of two physiological races of Saccharomyces cerevisiae. Lett Appl Microbiol 17:215–219. https://doi.org/10.1111/j.1472-765X.1993.tb01450.x
Fierro-Risco J, Rincón AM, Benítez T, Codón AC (2013) Overexpression of stress-related genes enhances cell viability and velum formation in Sherry wine yeasts. Appl Microbiol Biotechnol 97:6867–6881. https://doi.org/10.1007/s00253-013-4850-9
Fleet GH, Phaff HJ (1981) Fungal glucans-structure and metabolism. In: Tanner W, Loewns FA (eds) Encyclopedia of plant physiology new series, plant carbohydrates II. Verlag KG Springer, Berlin, pp 416–440
Gutiérrez P, Roldán A, Caro I, Pérez L (2010) Kinetic study of the velum formation by Saccharomyces cerevisiae (beticus ssp.) during the biological aging of wines. Process Biochem 45:493–499. https://doi.org/10.1016/j.procbio.2009.11.005
Ishigami M, Nakagawa Y, Hayakawa M, Iimura Y (2006) FLO11 is the primary factor in flor formation caused by cell surface hydrophobicity in wild-type flor yeast. Biosci Biotechnol Biochem 70:660–666. https://doi.org/10.1271/bbb.70.660
Kind T, Wohlgemuth G, Lee DY, Lu Y, Palazoglu M, Shahbaz S, Fiehn O (2009) FiehnLib: mass spectral and retention index libraries for metabolomics based on quadrupole and time-of-flight gas chromatography/mass spectrometry. Anal Chem 81:10038–10048. https://doi.org/10.1021/ac9019522
Krogan NJ, Dover J, Khorrami S, Greenblatt JF, Schneider J, Johnston M, Shilatifard A (2002) COMPASS, a histone H3 (Lysine 4) methyltransferase required for telomeric silencing of gene expression. J Biol Chem 277:10753–10755. https://doi.org/10.1074/jbc.C200023200
Legras JL, Moreno-García J, Zara S, Zara G, Garcia-Martinez T, Mauricio JC, Mannazzu I, Coi AL, Zeidan MB, Dequin S, Moreno J, Budroni M (2016) Flor Yeast: new perspectives beyond wine aging. Front Microbiol 7:1–11. https://doi.org/10.3389/fmicb.2016.00503
Li Y, Yan J, Kim I, Liu C, Huo K, Rao H (2010) Rad4 regulates protein turnover at a postubiquitylation step. Mol Biol Cell 21:177–185. https://doi.org/10.1091/mbc.E09-04-0305
López-Martínez G, Rodríguez-Porrata B, Margalef-Català M, Cordero-Otero R (2012) The Stf2p hydrophilin from Saccharomyces cerevisiae is required for dehydration stress tolerance. PLoS ONE 7:1–10. https://doi.org/10.1371/journal.pone.0033324
Lutfiyya LL, Johnston M (1996) Two zinc-finger-containing repressors are responsible for glucose repression of SUC2 expression. Mol Cell Biol 16:4790–4797
Martínez P, Pérez Rodríguez L, Benítez T (1997) Factors which affect velum formation by flor yeasts isolated from sherry wine. Syst Appl Microbiol 20:154–157. https://doi.org/10.1016/S0723-2020(97)80060-4
Mauricio JC, Moreno JJ, Ortega JM (1997) In vitro specific activities of alcohol and aldehyde dehydrogenases from two flor yeasts during controlled wine ageing. J Agric Food Chem 45:1967–1971. https://doi.org/10.1021/jf960634i
Mazur P, Morin N, Baginsky W, el-Sherbeini M, Clemas JA, Nielsen JB, Foor F (1995) Differential expression and function of two homologous subunits of yeast 1,3-beta-d-glucan synthase. Mol Cell Biol 15:5671–5681. https://doi.org/10.1128/MCB.15.10.5671
Medicherla B, Kostova Z, Schaefer A, Wolf DH (2004) A genomic screen identifies Dsk2p and Rad23p as essential components of ER-associated degradation. EMBO Rep 5:692–697. https://doi.org/10.1038/sj.embor.7400164
Moqtaderi Z, Struhl K (2004) Genome-wide occupancy profile of the RNA polymerase III machinery in Saccharomyces cerevisiae reveals loci with incomplete transcription complexes. Mol Cell Biol 24:4118–4127. https://doi.org/10.1128/MCB.24.10.4118-4127.2004
Moreno-García J, Raposo RM, Moreno J (2013) Biological aging status characterization of Sherry type wines using statistical and oenological criteria. Food Res Int 54:285–292. https://doi.org/10.1016/j.foodres.2013.07.031
Moreno-García J, García-Martínez T, Moreno J, Millán MC, Mauricio JC (2014) A proteomic and metabolomic approach for understanding the role of the flor yeast mitochondria in the velum formation. Int J Food Microbiol 172:21–29. https://doi.org/10.1016/j.ijfoodmicro.2013.11.030
Moreno-García J, García-Martínez T, Millán MC, Mauricio JC, Moreno J (2015a) Proteins involved in wine aroma compounds metabolism by a Saccharomyces cerevisiae flor-velum yeast strain grown in two conditions. Food Microbiol 51:1–9. https://doi.org/10.1016/j.fm.2015.04.005
Moreno-García J, García-Martínez T, Moreno J, Mauricio JC (2015b) Proteins involved in flor yeast carbon metabolism under biofilm formation conditions. Food Microbiol 46:25–33. https://doi.org/10.1016/j.fm.2014.07.001
Moreno-García J, Mauricio JC, Moreno J, García-Martínez T (2016) Stress responsive proteins of a flor yeast strain during the early stages of biofilm formation. Process Biochem 51:578–588. https://doi.org/10.1016/j.procbio.2016.02.011
Moreno-García J, Mauricio JC, Moreno J, García-Martínez T (2017) Differential proteome analysis of a flor yeast strain under biofilm formation. Int J Mol Sci 18:1–18. https://doi.org/10.3390/ijms18040720
Moreno-García J, Coi A, Zara G, García-Martínez T, Mauricio JC, Budroni M (2018) Study of the role of the covalently linked cell wall protein (Ccw14p) and yeast GlycoProtein (Ygp1p) within biofilm formation in a flor yeast strain. FEMS Yeast Res 18:1–5. https://doi.org/10.1093/femsyr/foy005
Peinado RA, Moreno JJ, Ortega JM, Mauricio JC (2003) Effect of gluconic acid consumption during simulation of biological aging of sherry wines by a flor yeast strain on the final volatile compounds. J Agric Food Chem 51:6198–6203. https://doi.org/10.1021/jf034512j
Perlman D, Raney P, Halvorson HO (1984) Cytoplasmic and secreted Saccharomyces cerevisiae invertase mRNAs encoded by one gene can be differentially or coordinately regulated. Mol Cell Biol 4:1682–1688
Rodríguez ME, Infante JJ, Mesa JJ, Rebordinos L, Cantoral JM (2013) Enological behaviour of biofilms formed by genetically-characterized strains of sherry flor yeast. Open Biotechnol J 7:23–29. https://doi.org/10.2174/1874070701307010023
Saccharomyces Genome Database. http://www.yeastgenome.org/. Accessed 2 Oct 2017
Vandenbosch D, De Canck E, Dhondt I, Rigole P, Nelis HJ, Coenye T (2013) Genomewide screening for genes involved in biofilm formation and miconazole susceptibility in Saccharomyces cerevisiae. FEMS Yeast Res 13:720–730. https://doi.org/10.1111/1567-1364.12071
Welker S, Rudolph B, Frenzel E, Hagn F, Liebisch G, Schmitz G, Scheuring J, Kerth A, Blume A, Weinkauf S, Haslbeck M, Kessler H, Buchner J (2010) Hsp12 is an intrinsically unstructured stress protein that folds upon membrane association and modulates membrane function. Mol Cell 39:507–520. https://doi.org/10.1016/j.molcel.2010.08.001
Zara S, Farris GA, Budroni M, Bakalinsky AT (2002) HSP12 is essential for biofilm formation by a Sardinian wine strain of Saccharomyces cerevisiae. Yeast 19:269–276. https://doi.org/10.1002/yea.831
Zara S, Bakalinsky AT, Zara G, Pirino G, Demontis MA, Budroni M (2005) FLO11-based model for air-liquid interfacial biofilm formation by Saccharomyces cerevisiae. Appl Environ Microbiol 71:2934–2939. https://doi.org/10.1128/AEM.71.6.2934-2939.2005
Zara G, Zara S, Pinna C, Marceddu S, Budroni M (2009) FLO11 gene length and transcriptional level affect biofilm-forming ability of wild flor strains of Saccharomyces cerevisiae. Microbiology 155:3838–3846. https://doi.org/10.1099/mic.0.028738-0
Zara S, Gross MK, Zara G, Budroni M, Bakalinsky AT (2010) Ethanol-independent biofilm formation by a flor wine yeast strain of Saccharomyces cerevisiae. Appl Environ Microbiol 76:4089–4091. https://doi.org/10.1128/AEM.00111-10
Zara G, Goffrini P, Lodi T, Zara S, Manazzu I (2012) FLO11 expression and lipid biosynthesis are required for air-liquid biofilm formation in a Saccharomyces cerevisiae flor strain. FEMS Yeast Res 12:864–866. https://doi.org/10.1111/j.1567-1364.2012.00831.x
Acknowledgements
Kind help of the staff at the Central Research Support Service (SCAI) of the university of Córdoba (Spain).
Funding
This work was supported by the “XXIII Programa Propio de Fomento de la Investigación 2018 UCO. MOD 4. Submodalidad 4.2. SINERGIAS” (XXIII.PP Mod. 4.2) from University of Cordoba (Spain) [J.C. Mauricio]; funding from the Spain’s Ministry of Education, Culture and Sport, Grants for training university teachers (FPU) [Jaime Moreno García].
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Moreno-García, J., Ogawa, M., Joseph, C.M.L. et al. Comparative analysis of intracellular metabolites, proteins and their molecular functions in a flor yeast strain under two enological conditions. World J Microbiol Biotechnol 35, 6 (2019). https://doi.org/10.1007/s11274-018-2578-5
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11274-018-2578-5