foodsystFood systemsПищевые системы2618-97712618-7272Федеральный научный центр пищевых систем им. В.М. Горбатова РАН10.21323/2618-9771-2025-8-4-488-497foodsyst-914Research ArticleСтатьиThe effect of lyophilization and cryoprotection on survival of Lactiplantibacillus plantarum strainsВлияние лиофилизации и криопротекции на выживаемость культур Lactiplantibacillus plantarumhttps://orcid.org/0000-0002-1108-3695СорокинаН. П.SorokinaN. P.

Сорокина Нинель Петровна — кандидат технических наук, ведущий научный сотрудник, отдел микробиологических исследований

152613, Ярославская область, Углич, ул. Красноармейский бульвар, 19 

Ninel P. Sorokina, Candidate of Technical Sciences, Leading Researcher, Department of Microbiological Research

19, Krasnoarmeysky Boulevard, Uglich, 152613, Yaroslavl Region

n.sorokina@fncps.ruhttps://orcid.org/0000-0002-5663-3662СмыковИ. Т.SmykovI. T.

Смыков Игорь Тимофеевич — доктор технических наук, главный научный сотрудник, отдел физико-химических исследований

152613, Ярославская область, Углич, ул. Красноармейский бульвар, 19 

Igor’ T. Smykov, Doctor of Technical Sciences, Chief Researcher, Department of Physico-chemical Research

19, Krasnoarmeysky Boulevard, Uglich, 152613, Yaroslavl Region

i.smykov@fncps.ruhttps://orcid.org/0000-0001-8251-992XКучеренкоИ. В.KucherenkoI. V.

Кучеренко Ирина Валентиновна — старший научный сотрудник, отдел микробиологических исследований

152613, Ярославская область, Углич, ул. Красноармейский бульвар, 19 

Irina V. Kucherenko, Senior Researcher, Department of Microbiological Research

19, Krasnoarmeysky Boulevard, Uglich, 152613, Yaroslavl Region

i.kucherenko@fncps.ruhttps://orcid.org/0000-0002-0710-3083КураеваЕ. В.KuraevaE. V.

Кураева Елена Вячеславовна — старший научный сотрудник, лаборатория селекции микроорганизмов

152613, Ярославская область, Углич, ул. Красноармейский бульвар, 19 

Elena V. Kuraeva, Senior Researcher, Microorganism Selection Laboratory

19, Krasnoarmeysky Boulevard, Uglich, 152613, Yaroslavl Region

e.kuraeva@fncps.ruhttps://orcid.org/0000-0001-9033-5928ДугановаА. Ю.DuganovaA. Yu.

Дуганова Анна Юрьевна — младший научный сотрудник, аспирант, лаборатория селекции микроорганизмов

152613, Ярославская область, Углич, ул. Красноармейский бульвар, 19 

Anna Yu. Duganova, Junior Researcher, Graduate Student, Microorganism Selection Laboratory

19, Krasnoarmeysky Boulevard, Uglich, 152613, Yaroslavl Region

a.duganova@fncps.ruВсероссийский научно-исследовательский институт маслоделия и сыроделияРоссияAll-Russian Scientific Research Institute of Butter and CheesemakingRussian Federation20252801202684488497Copyright © Sorokina N.P., Smykov I.T., Kucherenko I.V., Kuraeva E.V., Duganova A.Y., 20262026Сорокина Н.П., Смыков И.Т., Кучеренко И.В., Кураева Е.В., Дуганова А.Ю.Sorokina N.P., Smykov I.T., Kucherenko I.V., Kuraeva E.V., Duganova A.Y.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.fsjour.com/jour/article/view/914

Lyophilization has been recognized for many years as an effective method for storing collection cultures, bacterial starter cultures, and probiotics. Cryoprotectors are used to protect bacterial cells from stress during freezing and drying, and to increase their survival. The purpose of our study was to evaluate an effect of three freezing regimes before freeze-drying at temperatures of –20, –30, and –55 °C and solutions of two cryoprotectors, sucrose and trehalose, on the survival and preservation of the metabolic activity of mesophilic lactobacilli. The cultures were grown in 10 % skimmed milk, and in experimental versions, a 20 % cryoprotector solution was added to the culture in a 4:1 ratio. To determine an effect of freeze-drying conditions on bacterial survival and metabolic activity, complete two-factor, three-level experiments were developed and conducted. Two strains of the species Lactiplantibacillus plantarum from the VNIIMS collection, which are used in bacterial starter cultures for cheeses, were studied as representative microorganisms of lactobacilli. The results of assessing the viability of bacteria after lyophilization showed that both cryoprotectors increased the survival of cultures under all freezing conditions, but the best survival rates were obtained using trehalose. Strain-dependent effectiveness of lyophilization of the studied strains was established: in strain Lactiplantibacillus plantarum 28, the maximum survival was at a freezing temperature of –30 °C, in strain Lactiplantibacillus plantarum 37 — at a temperature of –55 °C with the addition of cryoprotector trehalose to both strains. Pre-freezing at –20 °C, also with trehalose, was the best method for preserving the metabolic activity of both cultures. It was shown that the greatest effectiveness of sucrose and trehalose as lyoprotectants during freeze-drying for both bacterial strains was found within the same pre-freezing temperature range of –30 °C. The identified strain-dependent effectiveness of lyophilization should be considered when selecting drying regimens for various collection cultures intended for long-term storage and requires an individual approach for each culture. The obtained results can also be used in freeze-drying of bacterial mass for the production of single-species bacterial starter cultures.

Лиофилизация на протяжении многих лет признается эффективным методом сохранения коллекционных культур, бактериальных заквасок и пробиотиков. Для защиты бактериальных клеток от стрессовых воздействий в процессе замораживания и сушки, для повышения их выживаемости используются различные криопротекторы. Целью исследования была оценка влияния трех режимов замораживания перед лиофильной сушкой при температуре минус 20, 30 и 55 °C и растворов двух криопротекторов — сахарозы и трегалозы на выживаемость и сохранение метаболической активности мезофильных лактобацилл. Культуры выращивали в 10 % обезжиренном молоке, в опытных вариантах к исследуемой культуре добавляли 20 % раствор криопротектора в соотношении 4:1. Для определения влияния условий лиофильной сушки на выживаемость и метаболическую активность бактерий, были разработаны и проведены полные двухфакторные трёхуровневые эксперименты. В качестве представительных микроорганизмов лактобацилл исследовали два штамма вида Lactiplantibacillus plantarum из коллекции ВНИИМС, которые используются в составе бактериальных заквасок для сыров. Результаты оценки жизнеспособности бактерий после лиофилизации показали, что оба криопротектора повышают выживаемость культур при всех режимах замораживания, но лучшие показатели выживаемости получены с использованием трегалозы. Установлена штамм-зависимость эффективности лиофилизации исследованных образцов: у штамма Lactiplantibacillus plantarum 28 максимальная выживаемость была выявлена при использовании температуры замораживания минус 30 °C, у штамма Lactiplantibacillus plantarum 37 — при температуре минус 55 °C с добавлением криопротектора трегалозы к обоим штаммам. Для сохранения метаболической активности обеих культур наилучшим было предварительное замораживание при температуре минус 20 °C и также с трегалозой. Показано, что наибольшая эффективность действия сахарозы и трегалозы в качестве лиопротекторов при лиофильной сушке, для обоих штаммов бактерий находится в одном и том же диапазоне температур предварительного замораживания близкой к минус 30 °C. Выявленная штамм-зависимость эффективности лиофилизации должна учитываться при выборе режимов сушки различных коллекционных культур, предназначенных для длительного хранения, и предполагает  индивидуальный подход к каждой культуре. Полученные результаты также могут быть использованы при лиофильной сушке бактериальной массы при производстве моновидовых бактериальных заквасок.

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