tatpipТехника и технология пищевых производствFood Processing: Techniques and Technology2074-94142313-1748Кемеровский государственный университет10.21603/2074-9414-2023-1-2415tatpip-10Research ArticleСтатьиSO2-резистентность как критерий отбора штаммов Saccharomyces cerevisiae для органического виноделияResistance to Sulfur Dioxide as a Criterion for Selecting Saccharomyces cerevisiae for Organic Winemakinghttps://orcid.org/0000-0002-7847-1246ПесковаИ. В.PeskovaI. V.

Ялта

Irina V. Peskova

Yalta

bioxim2012@mail.ruhttps://orcid.org/0000-0002-7847-1246ТанащукТ. Н.TanashchukT. N.

Ялта

Tatiana N. Tanashchuk

Yalta

https://orcid.org/0000-0003-0638-9187ОстроуховаЕ. В.OstroukhovaE. V.

Ялта

 Elena V. Ostroukhova

Yalta

https://orcid.org/0000-0002-8126-7596ЛутковаН. Ю.LutkovaN. Yu.

Ялта

Nataliya Yu. Lutkova

Yalta

https://orcid.org/0000-0001-6146-2151ВьюгинаМ. А.VyuginaM. A.

Ялта

Mariya A. Vyugina

Yalta

Всероссийский национальный научно-исследовательский институт виноградарства и виноделия «Магарач» РАНРоссияAll-Russian National Research Institute of Viticulture and Winemaking “Magarach” of the RASRussian Federation2023170820265316068Copyright © Пескова И.В., Танащук Т.Н., Остроухова Е.В., Луткова Н.Ю., Вьюгина М.А., 20262026Пескова И.В., Танащук Т.Н., Остроухова Е.В., Луткова Н.Ю., Вьюгина М.А.Peskova I.V., Tanashchuk T.N., Ostroukhova E.V., Lutkova N.Y., Vyugina M.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.fptt.ru/jour/article/view/10

Диоксид серы, используемый в виноделии как консервант и антиоксидант, негативно влияет на здоровье человека. Снижение доз диоксида серы возможно путем применения дрожжей с хорошей бродильной активностью, доминирующих при инокуляции в сусло, и низкой способностью синтезировать SO2 и SO2-связывающие вещества. Последнее связано с механизмами детоксикации SO2 дрожжами. Цель работы – изучение взаимосвязи устойчивости дрожжей к диоксиду серы и их способности синтезировать SO2 и ацетальдегид в процессе роста.

Исследовали 17 штаммов дрожжей рода Saccharomyces. Культивирование дрожжей осуществляли на установке CGQ на виноградном сусле до достижения стационарной фазы роста. Концентрацию диоксида серы определяли титриметри ческим методом, альдегидов – бисульфитным, сульфитоустойчивость дрожжей – по ростовой реакции на SO2 по технологии CGQ.

Установлено, что штаммы различались по степени SO2-резистентности, оцениваемой по увеличению продолжительности лаг-фазы в присутствии (100 мг/дм3) диоксида серы: первая группа (чувствительные) – 8 ч и более, вторая – 2–6 ч, третья (устойчивые) – без изменения. Выявлено (Wilks L = 0,228, α = 0,05), что в среде без SO2 чувствительные культуры отличались наибольшим значением минимального времени генерации в экспоненциальной фазе роста (5,3 ± 2,1 ч), устойчивые – наибольшим синтезом ацетальдегида (54,7 ± 11,1 мг/дм3) и диоксида серы (21,0 ± 10,3 мг/дм3), культуры второй группы – наименьшим содержанием в среде связанных форм SO2 (10,9 ± 4,2 мг/дм3), занимая по остальным показателям промежуточное положение.

В работе была показана возможность использования длительности адаптации дрожжей к SO2 для первичного отбора культур в эковиноделии. По совокупности физиологических и биохимических особенностей выделили перспективные культуры: из третьей группы – для производства вин с пониженным содержанием SO2, из второй – органических вин. Продолжение исследования направлено на расширение спектра штаммов дрожжей и уточнение диапазонов показателей для эффективного включения культур в виноделие.

Sulfur dioxide is a popular conserving agent and antioxidant in winemaking. Unfortunately, it is bad for human health. Some yeast strains can reduce the dose of sulfur dioxide. Such yeasts should have good fermentation activity and dominate when inoculated into grape must. In addition, it should not synthesize sulfur dioxide and SO2-binding substances. The synthesis of sulfur dioxide and carbonyl compounds by yeast is related to the mechanisms of sulfur dioxide detoxification. The research objective was to study the relationship between the resistance of yeast to sulfur dioxide and its ability to synthesize sulfur dioxide and acetaldehyde during growth.

The study featured 17 yeast strains of the genus Saccharomyces. The yeasts were cultivated on grape must in a CGQ device until the stationary growth phase. The concentration of free and bound forms of sulfur dioxide was determined by titration, while that of aldehydes was determined by bisulfite method. The sulfite resistance of strains was measured by the growth response of yeast cells to sulfur dioxide using CGQ technology.

Yeast strains differed in the degree of sulfur dioxide resistance. The samples were divided according to the increase in the lag phase time: by ≥ 8 h (sensitive), by 2–6 h, without changes (resistant). At Wilks L = 0.228 and α = 0.05, the sensitive cultures in a SO2-free medium had the highest value of minimal generation time in the exponential growth phase (5.3 ± 2.1 h).

The resistant samples demonstrated the highest synthesis of acetaldehyde (54.7 ± 11.1 mg/L) and sulfur dioxide (21.0 ± 10.3 mg/L). The second group cultures had the lowest content of SO2-bound forms in the medium (10.9 ± 4.2 mg/L) and were in an interposition in terms of other indicators.

The time it takes a yeast strain to adapt to sulfur dioxide can be used as a parameter for the primary culture selection in eco-winemaking. According to the physiological and biochemical profile, the resistant strains can be recommended for the production of SO2-low wines, while the samples from the second test group proved optimal for organic wines. Further research will expand the range of yeast strains and their indicators.

дрожжиCGQ технологиялаг-фазагенерациясинтезацетальдегиддиоксид серыэковиноделиеYeastsCGQ technologylag phasegenerationsynthesisacetaldehydesulfur dioxideeco-winemakingРабота выполнена на базе Всероссийского национального научно-исследовательского института виноградарства и виноделия «Магарач» РАН (ВННИИВиВ «Магарач» РАН) в рамках Государственного задания № 0833-2019-0022.The research was performed on the premises of the All-Russian National Research Institute of Viticulture and Winemaking “Magarach” of the RAS (“Magarach”) , public assignment No. 0833-2019-022.ReferencesGuerrero RF, Cantos-Villar E. Demonstrating the efficiency of sulphur dioxide replacements in wine: A parameter review. Trends in Food Science and Technology. 2015;42(1):27–43. https://doi.org/10.1016/j.tifs.2014.11.004Guerrero RF, Cantos-Villar E. Demonstrating the efficiency of sulphur dioxide replacements in wine: A parameter review. Trends in Food Science and Technology. 2015;42(1):27–43. https://doi.org/10.1016/j.tifs.2014.11.004Romano P, Ciani M, Fleet GH. Yeasts in the production of wine. New York: Springer; 2019. 515 р. https://doi.org/10.1007/978-1-4939-9782-4Romano P, Ciani M, Fleet GH. Yeasts in the production of wine. New York: Springer; 2019. 515 р. https://doi.org/10.1007/978-1-4939-9782-4Costanigro M, Appleby C, Menke SD. The wine headache: Consumer perceptions of sulfites and willingness to pay for non-sulfited wines. Food Quality and Preference. 2014;31:81–89. https://doi.org/10.1016/j.foodqual.2013.08.002Costanigro M, Appleby C, Menke SD. The wine headache: Consumer perceptions of sulfites and willingness to pay for non-sulfited wines. Food Quality and Preference. 2014;31:81–89. https://doi.org/10.1016/j.foodqual.2013.08.002Zara G, Nardi T. Yeast metabolism and its exploitation in emerging winemaking trends: From sulfite tolerance to sulfite reduction. Fermentation. 2021;7(2). https://doi.org/10.3390/fermentation7020057Zara G, Nardi T. Yeast metabolism and its exploitation in emerging winemaking trends: From sulfite tolerance to sulfite reduction. Fermentation. 2021;7(2). https://doi.org/10.3390/fermentation7020057Walker ME, Zhang J, Sumby KM, Lee A, Houlès A, Li S, et al. Sulfate transport mutants affect hydrogen sulfide and sulfite production during alcoholic fermentation. Yeast. 2021;38(6):367–381. https://doi.org/10.1002/yea.3553Walker ME, Zhang J, Sumby KM, Lee A, Houlès A, Li S, et al. Sulfate transport mutants affect hydrogen sulfide and sulfite production during alcoholic fermentation. Yeast. 2021;38(6):367–381. https://doi.org/10.1002/yea.3553Divol B, du Toit M, Duckitt E. Surviving in the presence of sulphur dioxide: Strategies developed by wine yeasts. Applied Microbiology and Biotechnology. 2012;95:601–613. https://doi.org/10.1007/s00253-012-4186-xDivol B, du Toit M, Duckitt E. Surviving in the presence of sulphur dioxide: Strategies developed by wine yeasts. Applied Microbiology and Biotechnology. 2012;95:601–613. https://doi.org/10.1007/s00253-012-4186-xGarcıa-Rıos E, Nuevalos M, Barrio E, Puig S, Guil- lamon JM. A new chromosomal rearrangement improves the adaptation of wine yeasts to sulfite. Environmental Microbiology. 2019;21(5):1771–1781. https://doi.org/10.1111/1462-2920.14586Garcıa-Rıos E, Nuevalos M, Barrio E, Puig S, Guil- lamon JM. A new chromosomal rearrangement improves the adaptation of wine yeasts to sulfite. Environmental Microbiology. 2019;21(5):1771–1781. https://doi.org/10.1111/1462-2920.14586Valero E, Tronchoni J, Morales P, Gonzalez R. Autophagy is required for sulfur dioxide tolerance in Saccharomyces cerevisiae. Microbial Biotechnology. 2019;13(2):599–604. https://doi.org/10.1111/1751-7915.13495Valero E, Tronchoni J, Morales P, Gonzalez R. Autophagy is required for sulfur dioxide tolerance in Saccharomyces cerevisiae. Microbial Biotechnology. 2019;13(2):599–604. https://doi.org/10.1111/1751-7915.13495Zimmer A, Durand C, Loira N, Durrens P, Sherman DJ, Marullo P. QTL dissection of lag phase in wine fermentation reveals a new translocation responsible for Saccharomyces cerevisiae adaptation to sulfite. PLoS ONE. 2014;9(1). https:// doi.org/10.1371/journal.pone.0086298Zimmer A, Durand C, Loira N, Durrens P, Sherman DJ, Marullo P. QTL dissection of lag phase in wine fermentation reveals a new translocation responsible for Saccharomyces cerevisiae adaptation to sulfite. PLoS ONE. 2014;9(1). https:// doi.org/10.1371/journal.pone.0086298Casalone E, Colella CM, Daly S, Gallori E, Moriani L, Polsinelli M. Mechanism of resistance to sulphite in Saccharomyces cerevisiae. Current Genetics. 1992;22(6):435–440. https://doi.org/10.1007/BF00326407Casalone E, Colella CM, Daly S, Gallori E, Moriani L, Polsinelli M. Mechanism of resistance to sulphite in Saccharomyces cerevisiae. Current Genetics. 1992;22(6):435–440. https://doi.org/10.1007/BF00326407ЦКП Коллекция микроорганизмов виноделия «Магарач». URL: http://magarach-institut.ru/kollekcija-mikroorganizmov-vinodelija-magarach (дата обращения: 10.06.2022).Magarach Common Use Center: Collection of Winemaking Microorganisms [Internet]. [cited 2022 Jun 10]. Available from: http://magarach-institut.ru/kollekcija-mikroorganizmov-vinodelija-magarach.Коллекция микроорганизмов виноделия института «Магарач» и ее роль в микробиологическом обеспечении отрасли / С. А. Кишковская [и др.] // Виноградарство и виноделие. 2016. Т. 46. С. 46–51.Kishkovskaya SA, Tanashchouk TN, Ivanova EV, Skorikova TK. Collection of microorganisms of winemaking of institute “Magarach” and role in microbiological supply industry. Viticulture and Winemaking. 2016;46:46–51. (In Russ.).Ivanova E. Study of acid-reducing ability of collection strains of wine yeast fungi-saccharomycetes. Norwegian Journal of Development of the International Science. 2021;(54–1):3–5. (In Russ.). https://doi.org/10.24412/3453-9875-2021-54-1-3-5Ivanova E. Study of acid-reducing ability of collection strains of wine yeast fungi-saccharomycetes. Norwegian Journal of Development of the International Science. 2021;(54–1):3–5. (In Russ.). https://doi.org/10.24412/3453-9875-2021-54-1-3-5Morgan SC, Haggerty JJ, Johnston B, Jiranek V, Durall DM. Response to sulfur dioxide addition by two commercial Saccharomyces cerevisiae strains. Fermentation. 2019;5(3). https://doi.org/10.3390/fermentation5030069Morgan SC, Haggerty JJ, Johnston B, Jiranek V, Durall DM. Response to sulfur dioxide addition by two commercial Saccharomyces cerevisiae strains. Fermentation. 2019;5(3). https://doi.org/10.3390/fermentation5030069Bruder S, Reifenrath M, Thomik T, Boles E, Herzod K. Parallelized online biomass monitoring in shake flasks enables efficient strain and carbon source dependent growth characterization of Saccharomyces cerevisiae. Microbial Cell Factories. 2016;15. https://doi.org/10.1186/s12934-016-0526-3Bruder S, Reifenrath M, Thomik T, Boles E, Herzod K. Parallelized online biomass monitoring in shake flasks enables efficient strain and carbon source dependent growth characterization of Saccharomyces cerevisiae. Microbial Cell Factories. 2016;15. https://doi.org/10.1186/s12934-016-0526-3Kuzmina SS, Kozubaeva LA, Egorova EYu, Kulushtayeva BM, Smolnikova FKh. Effect of berry extracts on Saccharomyces cerevisiae yeast. Food Processing: Techniques and Technology. 2021;51(4):819–831. (In Russ.). https://doi.org/10.21603/2074-9414-2021-4-819-831Kuzmina SS, Kozubaeva LA, Egorova EYu, Kulushtayeva BM, Smolnikova FKh. Effect of berry extracts on Saccharomyces cerevisiae yeast. Food Processing: Techniques and Technology. 2021;51(4):819–831. (In Russ.). https://doi.org/10.21603/2074-9414-2021-4-819-831Makarov AS, Lutkov IP. Yeast race effect on the quality of base and young sparkling wines. Foods and Raw Materials. 2021;9(2):290–301. https://doi.org/10.21603/2308-4057-2021-2-290-301Makarov AS, Lutkov IP. Yeast race effect on the quality of base and young sparkling wines. Foods and Raw Materials. 2021;9(2):290–301. https://doi.org/10.21603/2308-4057-2021-2-290-301García-Ríos E, Guillamón JM. Sulfur dioxide resistance in Saccharomyces cerevisiae: beyond SSU1. Microbial Cell. 2019;6(12):527–530. https://doi.org/10.15698/mic2019.12.699García-Ríos E, Guillamón JM. Sulfur dioxide resistance in Saccharomyces cerevisiae: beyond SSU1. Microbial Cell. 2019;6(12):527–530. https://doi.org/10.15698/mic2019.12.699Noble J, Sanchez I, Blondin B. Identification of new Saccharomyces cerevisiae variants of the MET2 and SKP2 genes con-trolling the sulfur assimilation pathway and the production of undesirable sulfur compounds during alcoholic fermentation. Microbial Cell Factories. 2015;14. https://doi.org/10.1186/s12934-015-0245-1Noble J, Sanchez I, Blondin B. Identification of new Saccharomyces cerevisiae variants of the MET2 and SKP2 genes con-trolling the sulfur assimilation pathway and the production of undesirable sulfur compounds during alcoholic fermentation. Microbial Cell Factories. 2015;14. https://doi.org/10.1186/s12934-015-0245-1Li E, Mira de Orduña R. Acetaldehyde metabolism in industrial strains of Saccharomyces cerevisiae inhibited by SO2 and cooling during alcoholic fermentation. OENO One. 2020;54(2):351–358. https://doi.org/10.20870/oeno-one.2020.54.2.2391Li E, Mira de Orduña R. Acetaldehyde metabolism in industrial strains of Saccharomyces cerevisiae inhibited by SO2 and cooling during alcoholic fermentation. OENO One. 2020;54(2):351–358. https://doi.org/10.20870/oeno-one.2020.54.2.2391Kishkovskaia SA, Tanashchuk TN, Avdanina DA, Eldarov MA, Ivanova EV, Shalamitskiy My, et al. Screening for promising yeast strains for sherry wine production using genetic and enological markers. Agricultural Biology. 2021;56(3):537–548. (In Russ.). https://doi.org/10.15389/agrobiology.2021.3.537engKishkovskaia SA, Tanashchuk TN, Avdanina DA, Eldarov MA, Ivanova EV, Shalamitskiy My, et al. Screening for promising yeast strains for sherry wine production using genetic and enological markers. Agricultural Biology. 2021;56(3):537–548. (In Russ.). https://doi.org/10.15389/agrobiology.2021.3.537engLi E, Mira de Orduña R. Acetaldehyde kinetics of enological yeast during alcoholic fermentation in grape must. Journal of Industrial Microbiology and Biotechnology. 2017;44(2):229–236. https://doi.org/10.1007/s10295-016-1879-7Li E, Mira de Orduña R. Acetaldehyde kinetics of enological yeast during alcoholic fermentation in grape must. Journal of Industrial Microbiology and Biotechnology. 2017;44(2):229–236. https://doi.org/10.1007/s10295-016-1879-7Peskova IV, Ostroukhova EV, Zaitseva OV, Lutkova NYu, Vyugina MA. The role of technological factors in the formation of SO2-binding complex of base wines. Magarach. Viticulture and Vinemaking. 2021;23(1):83–90. https://doi.org/10.35547/IM.2021.96.76.014Peskova IV, Ostroukhova EV, Zaitseva OV, Lutkova NYu, Vyugina MA. The role of technological factors in the formation of SO2-binding complex of base wines. Magarach. Viticulture and Vinemaking. 2021;23(1):83–90. https://doi.org/10.35547/IM.2021.96.76.014

The authors declare that there are no conflicts of interest present.