Abstract
The genetic improvement of winemaking yeasts is a virtually infinite process, as the design of new strains must always cope with varied and ever-evolving production contexts. Good wine yeasts must feature both good primary traits, which are related to the overall fermentative fitness of the strain, and secondary traits, which provide accessory features augmenting its technological value. In this context, the superiority of “blind,” genetic improvement techniques, as those based on the direct selection of the desired phenotype without prior knowledge of the genotype, was widely proven. Blind techniques such as adaptive evolution strategies were implemented for the enhancement of many traits of interest in the winemaking field. However, these strategies usually focus on single traits: this possibly leads to genetic tradeoff phenomena, where the selection of enhanced secondary traits might lead to sub-optimal primary fermentation traits. To circumvent this phenomenon, we applied a multi-step and strongly directed genetic improvement strategy aimed at combining a strong fermentative aptitude (primary trait) with an enhanced production of glutathione (secondary trait). We exploited the random genetic recombination associated to a library of 69 monosporic clones of strain UMCC 855 (Saccharomyces cerevisiae) to search for new candidates possessing both traits. This was achieved by consecutively applying three directional selective criteria: molybdate resistance (1), fermentative aptitude (2), and glutathione production (3). The strategy brought to the selection of strain 21T2-D58, which produces a high concentration of glutathione, comparable to that of other glutathione high-producers, still with a much greater fermentative aptitude.
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References
Ambroset C, Petit M, Brion C, Sanchez I, Delobel P, Guérin C, Chiapello H, Nicolas P, Bigey F, Dequin S, Blondin B (2011) Deciphering the molecular basis of wine yeast fermentation traits using a combined genetic and genomic approach. G3 Genes Genom Gene 1:263–281. https://doi.org/10.1534/g3.111.000422
Baleiras Couto MM, Eijsma B, Hofstra H, Huis in’t Veld JH, van der Vossen JM (1996) Evaluation of molecular typing techniques to assign genetic diversity among Saccharomyces cerevisiae strains. Appl Environ Microbiol 62(1):41–46
Bisson LF (1999) Stuck and sluggish fermentations. Am J Enol Vitic 50:107–119
Bonciani T, Solieri L, De Vero L, Giudici P (2016) Improved wine yeasts by direct mating and selection under stressful fermentative conditions. Eur Food Res Technol 6(6):899–910. https://doi.org/10.1007/s00217-015-2596-6
Cacho J, Castells JE, Esteban A, Laguna B, Sagristá N (1995) Iron, copper, and manganese influence on wine oxidation. Am J Enol Vitic 46:380–384
Çakar ZP, Seker UOS, Tamerler C, Sonderegger M, Sauer U (2005) Evolutionary engineering of multiple-stress resistant Saccharomyces cerevisiae. FEMS Yeast Res 5(6-7):569–578. https://doi.org/10.1016/j.femsyr.2004.10.010
Çakar ZP, Turanli-Yildiz B, Alkim C, Yilmaz Ü (2012) Evolutionary engineering of Saccharomyces cerevisiae for improved industrially important properties. FEMS Yeast Res 12(2):171–182. https://doi.org/10.1111/j.1567-1364.2011.00775.x
De Vero L, Solieri L, Giudici P (2011) Evolution-based strategy to generate non-genetically modified organisms Saccharomyces cerevisiae strains impaired in sulfate assimilation pathway. Lett Appl Microbiol 53(5):572–575. https://doi.org/10.1111/j.1472-765X.2011.03140.x
De Vero L, Bonciani T, Verspohl A, Mezzetti F, Giudici P (2017) High-glutathione producing yeasts obtained by genetic improvement strategies: a focus on adaptive evolution approaches for novel wine strains. AIMS Microbiol 3(2):155–170. https://doi.org/10.3934/microbiol.2017.2.155
Dragosits M, Mattanovich D (2013) Adaptive laboratory evolution—principles and applications for biotechnology. Microb Cell Factories 12(1):1–17. https://doi.org/10.1186/1475-2859-12-64
Dubourdieu D, Lavigne V (2004) The role of glutathione on the aromatic evolution of dry white wine. Vinidea Net - Wine Internet Tech J 2:1–9
Fleet GH (2003) Yeast interactions and wine flavour. Int J Food Microbiol 86(1-2):11–22. https://doi.org/10.1016/S0168-1605(03)00245-9
García-Esparza MA, Capri E, Pirzadeh P, Trevisan M (2006) Copper content of grape and wine from Italian farms. Food Addit Contam 23(3):274–280. https://doi.org/10.1080/02652030500429117
Giudici P, Kunkee RE (1994) The effect of nitrogen deficiency and sulfur-containing amino acids on the reduction of sulfate to hydrogen sulfide by wine yeasts. Am J Enol Vitic 45:107–112
Giudici P, Zambonelli C (1992) Criteri di selezione dei lieviti per enologia. Vignevini 9:29–34
Giudici P, Solieri L, Pulvirenti AM, Cassanelli S (2005) Strategies and perspectives for genetic improvement of wine yeasts. Appl Microbiol Biotechnol 66(6):622–628. https://doi.org/10.1007/s00253-004-1784-2
Gobbi M, De Vero L, Solieri L, Comitini F, Oro L, Giudici P, Ciani M (2014) Fermentative aptitude of non-Saccharomyces wine yeast for reduction in the ethanol content in wine. Eur Food Res Technol 239:41–48. https://doi.org/10.1007/s00217-014-2187-y
Grant CM, MacIver FH, Dawes IW (1996) Glutathione is an essential metabolite required for resistance to oxidative stress in the yeast Saccharomyces cerevisiae. Curr Genet 29(6):511–515. https://doi.org/10.1007/BF02426954
Hara S, Iimura Y, Otsuka K (1980) Breeding of useful killer wine yeasts. Am J Enol Vitic 31:28–33
Hoffman CS, Winston F (1987) A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene 57(2-3):267–272. https://doi.org/10.1016/0378-1119(87)90131-4
Kritzinger EC, Bauer FF, Du Toit WJ (2013) Role of glutathione in winemaking. J Agric Food Chem 61(2):269–277. https://doi.org/10.1021/jf303665z
Legras JL, Karst F (2003) Optimisation of interdelta analysis for Saccharomyces cerevisiae strain characterisation. FEMS Microbiol Lett 221(2):249–255. https://doi.org/10.1016/S0378-1097(03)00205-2
Marullo P, Bely M, Masneuf-Pomarede I, Aigle M, Dubourdieu D (2004) Inheritable nature of enological quantitative traits is demonstrated by meiotic segregation of industrial wine yeast strains. FEMS Yeast Res 4(7):711–719. https://doi.org/10.1016/j.femsyr.2004.01.006
Mezzetti F, De Vero L, Giudici P (2014) Evolved Saccharomyces cerevisiae wine strains with enhanced glutathione production obtained by an evolution-based strategy. FEMS Yeast Res 14(6):977–987. https://doi.org/10.1111/1567-1364.12186
Mezzetti F, Fay JC, Giudici P, De Vero L (2017) Genetic variation and expression changes associated with molybdate resistance from a glutathione producing wine strain of Saccharomyces cerevisiae. PLoS One 12(7):e0180814. https://doi.org/10.1371/journal.pone.0180814
Mortimer RK, Romano P, Suzzi G, Polsinelli M (1994) Genome renewal: a new phenomenon revealed from a genetic study of 43 strains of Saccharomyces cerevisiae derived from natural fermentation of grape musts. Yeast 10(12):1543–1552. https://doi.org/10.1002/yea.320101203
Patzschke A, Steiger MG, Holz C, Lang C, Mattanovich D, Sauer M (2015) Enhanced glutathione production by evolutionary engineering of Saccharomyces cerevisiae strains. Biotechnol J 10(11):1719–1726. https://doi.org/10.1002/biot.201400809
Penninckx M (2000) A short review on the role of glutathione in the response of yeasts to nutritional, environmental, and oxidative stresses. Enzym Microbiol Technol 26(9-10):737–742. https://doi.org/10.1016/S0141-0229(00)00165-4
Pretorius IS (2000) Tailoring wine yeast for the new millennium: novel approaches to the ancient art of winemaking. Yeast 16(8):675–729. https://doi.org/10.1002/1097-0061(20000615)16:8<675::AID-YEA585>3.0.CO;2-B
Quatrini P, Marineo S, Puglia AM, Restuccia C, Randazzo CL, Spagna G, Barbagallo RN, Palmeri R, Giudici P (2008) Partial sequencing of the β-glucosidase-encoding gene of yeast strains isolated from musts and wines. Ann Microbiol 58(3):503–508. https://doi.org/10.1007/BF03175549
Rainieri S, Zambonelli C, Giudici P, Castellari L (1998) Characterisation of thermotolerant Saccharomyces cerevisiae strains. Biotechnol Lett 20(6):543–547. https://doi.org/10.1023/A:1005389309527
Regodón JA, Peréz F, Valdés ME, De Miguel C, Ramírez M (1997) A simple and effective procedure for selection of wine yeast strains. Food Microbiol 14(3):247–254. https://doi.org/10.1006/fmic.1996.0091
Romano P, Soli MG, Suzzi G, Grazia L, Zambonelli C (1985) Improvement of a wine Saccharomyces cerevisiae strain by a breeding program. Appl Environ Microbiol 50:1064–1067
Roussis IG, Papadopoulou D, Sakarellos-Daitsiotis M (2009) Protective effect of thiols on wine aroma volatiles. Open Food Sci J 3:98–102
Sala C, Fort F, Busto O, Zamora F, Arola L, Guasch J (1996) Fate of some common pesticides during vinification process. J Agric Food Chem 44(11):3668–3671. https://doi.org/10.1021/jf960218y
Salvadó Z, Arroyo-López FN, Barrio E, Querol A, Guillamón JM (2011) Quantifying the individual effects of ethanol and temperature on the fitness advantage of Saccharomyces cerevisiae. Food Microbiol 28(6):1155–1161. https://doi.org/10.1016/j.fm.2011.03.008
Schütz M, Gafner J (1994) Dynamics of the yeast strain population during spontaneous alcoholic fermentation determined by CHEF gel electrophoresis. Lett Appl Microbiol 19(4):253–257. https://doi.org/10.1111/j.1472-765X.1994.tb00957.x
Simpson RF (1982) Factors affecting oxidative browning of white wine. Vitis 21:233–239
Sipiczki M (2011) Diversity, variability and fast adaptive evolution of the wine yeast (Saccharomyces cerevisiae) genome—a review. Ann Microbiol 61(1-2):85–93. https://doi.org/10.1016/S0168-1605(03)00245-9
Sipiczki M, Romano P, Capece A, Paraggio M (2004) Genetic segregation of natural Saccharomyces cerevisiae strains derived from spontaneous fermentation of Aglianico wine. J Appl Microbiol 96(5):1169–1175. https://doi.org/10.1111/j.1365-2672.2004.02254.x
Sonni F, Clark AC, Prenzler PD, Riponi C, Scollary GR (2011a) Antioxidant action of glutathione and the ascorbic acid/glutathione pair in a model white wine. J Agric Food Chem 59(8):3940–3949. https://doi.org/10.1021/jf104575w
Sonni F, Moore EG, Clark AC, Chinnici F, Riponi C, Scollary GR (2011b) Impact of glutathione on the formation of methylmethine-and carboxymethine-bridged (+)-catechin dimers in a model wine system. J Agric Food Chem 59(13):7410–7418. https://doi.org/10.1021/jf200968x
Swiegers J, Bartowsky E, Henschke PA, Pretorius IS (2005) Yeast and bacterial modulation of wine aroma and flavour. Aust J Grape Wine Res 11(2):139–173. https://doi.org/10.1111/j.1755-0238.2005.tb00285.x
Swinnen S, Schaerlaekens K, Pais T, Claesen J, Hubmann G, Yang Y, Demeke M, Foulquié-Moreno MR, Goovaerts A, Souvereyns K, Clement L, Dumortier F, Thevelein JM (2012) Identification of novel causative genes determining the complex trait of high ethanol tolerance in yeast using pooled-segregant whole-genome sequence analysis. Genome Res 22(5):975–984. https://doi.org/10.1101/gr.131698.111
Tirelli A, Fracassetti D, De Noni I (2010) Determination of reduced cysteine in oenological cell wall fractions of Saccharomyces cerevisiae. J Agric Food Chem 58(8):4565–4570. https://doi.org/10.1021/jf904047u
Van Leeuwen C, Darriet P (2016) The impact of climate change on viticulture and wine quality. J Wine Econ 11(01):150–167. https://doi.org/10.1017/jwe.2015.21
Verspohl A, Solieri L, Giudici P (2017) Exploration of genetic and phenotypic diversity within Saccharomyces uvarum for driving strain improvement in winemaking. Appl Microbiol Biotechnol 101(6):2507–2521. https://doi.org/10.1007/s00253-016-8008-4
Winkler JD, Kao KC (2014) Recent advances in the evolutionary engineering of industrial biocatalysts. Genomics 104(6):406–411. https://doi.org/10.1016/j.ygeno.2014.09.006
Wysocki R, Tamás MJ (2010) How Saccharomyces cerevisiae copes with toxic metals and metalloids. FEMS Microbiol Rev 34(6):925–951. https://doi.org/10.1111/j.1574-6976.2010.00217.x
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This research project was financially supported by the AEB Group (Brescia, Italy).
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Bonciani, T., De Vero, L., Mezzetti, F. et al. A multi-phase approach to select new wine yeast strains with enhanced fermentative fitness and glutathione production. Appl Microbiol Biotechnol 102, 2269–2278 (2018). https://doi.org/10.1007/s00253-018-8773-3
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DOI: https://doi.org/10.1007/s00253-018-8773-3