spfpХранение и переработка сельхозсырьяStorage and Processing of Farm Products2072-96692658-767XРОСБИОТЕХ10.36107/spfp.2020.333spfp-147Research ArticleФИЗИЧЕСКИЕ И ХИМИЧЕСКИЕ МЕТОДЫ ПЕРЕРАБОТКИ СЕЛЬХОЗПРОДУКЦИИPHYSICAL AND CHEMICAL METHODS OF FARM RAW MATERIAL PROCESSINGВлияние температурного воздействия на концентрацию лактоферрина в коровьем молокеTemperature Impact on Lactoferrin Concentration in Bovine MilkДонскаяГ. А.DonskayaG. A.

Донская Галина Андреевна

115093, Москва, ул. Люсиновская, д.35, корп.7

Galina A. Donskaya

35/7, Ljusinovskaya Str., Moscow, 115093 

g_donskaya@vnimi.orgЧумаковаИ. В.ChumakovaI. V.

Чумакова Ирина Валерьевна

115093, Москва, ул. Люсиновская, д.35, корп.7

Irina V. Chumakova

35/7, Ljusinovskaya Str., Moscow, 115093 

e_chumakova@vnimi.orgФГАНУ «ВНИМИ»РоссияFGANU “VMINI”Russian Federation202013052021032030Copyright © Донская Г.А., Чумакова И.В., 20212021Донская Г.А., Чумакова И.В.Donskaya G.A., Chumakova I.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.spfp-mgupp.ru/jour/article/view/147

Объектом настоящего исследования являлся сывороточный протеин лактоферрин (ЛФ), который обладает защитными функциями и признан важным компонентом врожденного иммунитета организма. Этот белок проявляет антимикробные, противовирусные свойства, регулирует воспалительный процесс и участвует в активации иммунных клеток. ЛФ определяется в различных биологических жидкостях, но большая его концентрация показана в молозиве и молоке в составе сывороточных белков. Целью настоящей работы является выделение ЛФ из нативного (сырого) и термически обработанного молока для установления влияния температурных режимов на концентрацию этого белка. Объекты исследования: лактоферрин; сырое термически необработанное молоко, полученное из индивидуального хозяйства; пастеризованное молоко. Лактоферрин выделяли из обезжиренного молока с помощью ионообменной хроматографии. Анализы образцов ЛФ проводили методом ВЭЖХ. Концентрацию белка, выделенного из молока, определяли колориметрическим методом по Брэдфорд. Установили, что в молоке, пастеризованном при температуре (78±2)°С с выдержкой 20 сек., концентрация ЛФ, в сравнении с нативным молоком, снижалась до 29,1%. При температурном воздействии (92±1)°С с выдержкой 2 мин. концентрация ЛФ составляла менее 1% от концентрации ЛФ в нативном молоке. Очевидно, что снижение температуры пастеризации молока ниже 78°С будет способствовать большему сохранению сывороточного белка ЛФ, однако при этом потребуется длительная временная выдержка, которая приведёт к изменению физико-химических показателей молока. В связи с высокой термолабильностью сывороточного белка, целесообразно рассмотреть технологии деконтаминации молока без применения высоких температур.

The focus of current research was Lactoferrin (LF), whey protein that possesses protective functions and is regarded as an important component of innate immune system. This protein demonstrates antimicrobial and antiviral properties, regulates inflammation and participates in immune cells activation. LF is present in various biological fluids but most of it accumulates in whey proteins of colostrum and milk. The objective of this study was the isolation of LF from native (raw) and heat-treated milk to identify impact of various degrees of heat treatment on LF concentration. The object of research – Lactoferrin; raw unprocessed milk obtained from individual farm; pasteurized milk. Lactoferrin was isolated from skim milk with Ion-exchange Chromatography. Lactoferrin sample analysis was conducted with HPLC. Concentration of protein isolated from milk was done by Bredford calorimetric method. Results demonstrated that in milk, pasteurized at (78±2)0С with holding time 20 sec, LF concentration decreased and reached 29.1% when compared with native milk. Lactoferrin concentration at (92±1)0С and holding time 2 min was less than 1% of LF concentration in native milk. It is clear that keeping pasteurization temperature below 780С will promote higher LF stability, however, it will require longer holding time that will impact physico-chemical parameters of milk. Based on high heat resistance of whey protein, it is advisable to evaluate milk decontamination technology without significant heat stress application.

лактоферринхроматографиясырое молокотермически обработанное молокоlactoferrinchromatographyraw milkheat-treated milkReferencesAcафов В.А., Танькова Н.Л., Искакова Е.Л., Харитонов В.Д., Курченко В.П., Головач Т.Н. Некоторые аспекты регулирования микробиологического состава молозива // Пищевая индустрия. 2019. № 4(42). С. 20–25. https://doi.org/10.24411/9999-008A-2019-10015Asafov V.A., Tanjkova N.L., Iskakova E.L., Kharitonov V.D., Kurchenko V.P., Golovach T.N. Nekotorye aspecty reulirovanija mikrobiologicheskogo sostava moloziva [Some aspect of microbiological milk composition regulation]. Pischevaja industrija [Food industry], 2019, vol. 42, no. 4, pp. 20-25. https://doi.org/10.24411/9999-008A-2019-10015Зобкова З.С., Мишина А.В. Наночастицы лактоферрина в производстве функциональных молочных продуктов // Хранение и переработка с/х сырья. 2009. № 12. С. 49–52.Komolova G.S, Tikhomirova N.S., Ionova I.I., Komolov S.A. Lactoferrin korovjego moloka [Bovine lactoferrin]. Molochnaja promyshlennost [Dairy industry], 2011, no. 7, pp. 70-71.Зобкова З.С., Мишина А.В., Бегунова А.В. Антимикробные свойства лактоферрина // Молочная промышленность. 2007. № 10. с. 60.Kostevic, V.A., Sokolov A.V., Zaharova E.T., Vasiljev V.B. Analyz soderzhanija I nasyschennosti zhelezom lactoferrina v molokee zhenschin s pervogo dnja b lj 5 let lactacii. [Composition analysis and satiety level of lactoferrin with iron and copper in human milk from the first day and up to 5 years of lactation]. Medicinskij akademicheskij zhurnal [Medical academic journal], 2014, vol. 14, part 1, pp. 80-86.Зобкова З.С., Мишина А.В., Бегунова А.В. О бифидогенных свойствах лактоферрина // Молочная промышленность. 2008. № 7. С. 64–65.Titov E.I., Tikhomirova N.A., Ionova I.I. Vydelenie I izuchenie zhelezosvjzyvajuschei sposobnosti lactoferrina korovkjego moloka [Isolation and research of iron-binding potential of bovine milk lactoferrin]. Voprosy pitanija [Nutrition issues], 2019, vol. 88, no. 1, pp. 91-95.Зобкова З.С., Мишина А.В., Смолянинов В.В., Шехватова Г.В. Технология экстрагирования лактоферрина из сырого молока // Молочная промышленность. 2009. № 12. с. 21–22.Zobkova Z.A., Mishina A.V. Nanochasticy lactoferrina v proisvodstve funkcionaljnyh molochnyh productov [Nanoparticles of lactoferrin in production of functional dairy products]. Hranenie i pererabotka selkhozsyrya cyrja [Storage and processing of farm products], 2009, no 12, pp. 49-52.Зобкова З.С., Федотова, О. Б., Фурсова Т.П., Зенина Д.В., Гаврилина А.Д., Шелагинова И.Р. Исследование антибактериальных свойств лактоферрина в сметане // Научное обеспечение молочной промышленности: сборник трудов. М.: ФГБНУ «ВНИМИ», Издательство «Франтера», 2016. С. 91–100.Zobkova Z.S., Fedotova O.B. Fursova T.P., Zenina D.V., Gavrilina A.D., Shelaginova I.R. Issledovanije antibacterialjnyh svojstv lactoferrina v smetane: sbornik Trudov VNIMI [Research of antibacterial lactoferrin properties in sour cream: Collection of works of All-Russian Research Institute of Dairy Industry]. Moscow: FGBNU “VNIMI”, Izdatel’stvo “Frantera”, 2016, pp. 1-100.Комолова Г.С., Тихомирова Н.С., Ионова И.И., Комолов С.А. Лактоферрин коровьего молока // Молочная промышленность. 2011. № 7. С. 70–71.Zobkova Z.S., Mishina A.V., Begunova A.V. Antimikrobnyje svoestva lactoferrina [Antimicrobial properties of lactoferrin]. Molochnaya promyshlennost [Dairy industry], 2007, no. 10, pp. 60.Костевич В.А., Соколов А.В., Захарова Е.Т., Васильев В.Б. Анализ содержания и насыщенности железом и медью лактоферрина в молоке у женщин с первого дня и до 5 лет лактации // Медицинский академический журнал. 2014. T. 14, ч. 1. С. 80–86.Zobkova Z.S., Mishina A.V., Begunova A.V. O bifidogennyh svojstvah lactoferrina [About bifidogenich properties of lactoferrin]. Molochnaya promyshlennost [Dairy industry], 2008, no. 7, pp. 64-65.Титов Е.И., Тихомирова Н.А., Ионова И.И. Выделение и изучение железосвязывающей способности лактоферрина коровьего молока // Вопросы питания. 2019. Т. 88, № 1. С. 91–95. https://doi.org/10.24411/0042-8833-2019-10011Zobkova Z.S., Mishina A.V., Smoljaninov V.V., Shekvatova G.V. Tehnologija ekstragirovanija lactoferrina iz syrogo moloka [Lactoferrin extraction technology from raw milk]. Molochnaja promyshlennost [Dairy industry], 2009, no. 12, pp. 21-22.Bradford M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding // Analytical Biochemistry. 1976. Vol. 72, issues 1–2. P. 248–254. https://doi.org/10.1006/abio.1976.9999Bradford M.M. A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding. Analytical Biochemistry, 1976, vol. 72, issues 1-2, pp. 248-254. https://doi.org/10.1006/abio.1976.9999Brick T., Ege M., Boeren S., Bock, A., von Mutius E., Vervont J., Hettinga K. Effect of processing intensity on immunologically active bovine milk serum proteins // Nutrients. 2017. Vol. 9, issue 9. P. 1–14. https://doi.org/10.3390/nu9090963Brick T., Ege M., Boeren S., Bock, A., von Mutius E., Vervont J., Hettinga K. Effect of Processing Intensity on Immunologically Active Bovine Milk Serum Proteins. Nutrients, 2017, vol. 9, issue 9, p. 1-14. https://doi.org/10.3390/nu9090963Chow L.M., Subbaraman L.N., Sheardown H. Kinetics of in vitro Lactoferrin deposition on Silicone hydrogel and FDA group II and group IV hydrogel contact lens materials // Journal of Biomaterials Science, Polymer Edition. 2009. Vol. 20, issue 1. P. 71–82. https://doi.org/10.1163/156856208X393509Chow L.M., Subbaraman L.N., Sheardown H. Kinetics of in Vitro Lactoferrin Deposition on Silicone Hydrogel and Fda Group II and Group IV Hydrogel Contact Lens Materials. Journal of Biomaterials Science, Polymer Edition, 2009, vol 20, issue 1, pp. 71-82. https://doi.org/10.1163/156856208X393509Dissanayake, Ramchandran M., Donkor L., Vasiljevic O.T. Denaturation of whey proteins as function of heat, Ph and protein concentration // International Dairy Journal. 2013. Vol. 31, issue 2. P. 93–99. https://doi.org/10.1016/j.idairyj.2013.02.002Dissanayake, Ramchandran M., Donkor L., Vasiljevic O.T. Denaturation of Whey Proteins as Function of Heat, Ph And Protein Concentration. International Dairy Journal, 2013, vol. 31, issue 2, pp. 93-99. https://doi.org/10.1016/j.idairyj.2013.02.002Donovan S. Role of human milk components in gastrointestinal development: Current knowledge and future needs // The Journal of Pediatrics. 2006. Vol. 149, issue 5. P. 49–61. https://doi.org/10.1016/j.jpeds.2006.06.052Donovan S. Role of Human Milk Components in Gastrointestinal Development: Current Knowledge and Future Needs. The Journal of Pediatrics, 2006, vol. 149, issue 5, pp. 49-61. https://doi.org/10.1016/j.jpeds.2006.06.052Dritsakou K., Liosis G., Valsami G., Polychronopoulos E., Skouroliakou M. Improved outcomes of feeding low birth weight infants with predominantly raw human milk versus donor banked milk and formula // Journal of Maternal-Fetal & Neonatal Medicine. 2016. Vol. 29, issue 7. P. 1131–1138. https://doi.org/10.3109/14767058.2015.1038232Dritsakou K., Liosis G., Valsami G., Polychronopoulos E., Skouroliakou M. Improved Outcomes of Feeding Low Birth Weight Infants with Predominantly Raw Human Milk Versus Donor Banked Milk and Formula. Journal of Maternal-Fetal & Neonatal Medicine, 2016, vol. 29, issue 7, pp. 1131-1138. https://doi.org/10.3109/14767058.2015.1038232Fang B., Zhang M., Jiang T., Guo H., Zheng F. Bovine Lactoferrin binds oleic acid to form an antitumor complex similar to hamlet //Biochimica et Biophysica Acta (BBA) – Molecular and Cell Biology of Lipids. 2014. Vol. 1841, issue 4. P. 535– 543. https://doi.org/10.1016/j.bbalip.2013.12.008Fang B., Zhang M., Jiang T., Guo H., Zheng F. Bovine Lactoferrin Binds Oleic Acid to form an Antitumor Complex Similar to Hamlet. Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, 2014, vol. 1841, issue 4, pp. 535-543. https://doi.org/10.1016/j.bbalip.2013.12.008Gonzales-Chavez S.A., Arevalo-Gallegos S., RasconGruz Q. Lactoferrin structure, function and application // International Journal of Antimicrobial Agents. 2009. Vol. 33, issue 4. P. 301.e1–301.e8. https://doi.org/10.1016/j.ijantimicag.2008.07.020Gonzales-Chavez S.A., Arevalo-Gallegos S., RasconGruz Q. Lactoferrin Structure, Function and Application. International Journal of Antimicrobial Agents, 2009, vol. 33, issue 4, pp. 301.e1-301.e8. https://doi.org/10.1016/j.ijantimicag.2008.07.020Jenssen H., Hancock R. Antimicrobial properties of Lactoferrin // Biochimie. 2009. Vol. 91, issue 1. P. 19–29. https://doi.org/10.1016/j.biochi.2008.05.015Jenssen H., Hancock R. Antimicrobial Properties of Lactoferrin. Biochimie, 2009, Vol. 91, issue 1, pp. 19- 29. https://doi.org/10.1016/j.biochi.2008.05.015Kong Y. Y., Liu M., Di W., Wang C., Du M., Zhang L. W. Purification and identification of lactoferrin from bovine milk // Advanced Materials Research. 2012. Vol. 524–527. P. 2290–2293. https://doi.org/10.4028/www.scientific.net/AMR.524-527.2290Kong, Y. Y., Liu, M., Di, W., Wang, C., Du, M., Zhang, L. W. Purification and Identification of Lactoferrin from Bovine Milk. Advanced Materials Research, 2012, vol. 524-527, pp. 2290-2293. https://doi.org/10.4028/www.scientific.net/AMR.524-527.2290Lorget F., Clough J., Oliveira M., Daury M., Sabokbar A., Offord E. Lactoferrin reduces in vitro osteoclast differentiation and resorbing activity // Biochemical and Biophysical Research Communications. 2002. Vol. 296, issue 2. P. 261–266. https://doi.org/10.1016/s0006-291x(02)00849-5Lorget F., Clough J., Oliveira M., Daury M., Sabokbar A., Offord E. Lactoferrin Reduces in Vitro Osteoclast Differentiation and Resorbing Activity. Biochemical and Biophysical Research Communications, 2002, vol. 296, issue 2, pp. 261-266. https://doi.org/10.1016/s0006-291x(02)00849-5Loss G., Depner M., Ulfman L.H., van Neerven R.J., Hose A.J. Consumption of unprocessed cow´s milk protects infants from common respiratory infections // Journal of Allergy Clinical Immunology. 2015. Vol. 135, issue 1. P. 56–62. https://doi.org/10.1016/j.jaci.2014.08.044Loss G., Depner M., Ulfman L.H., vanNeerven R.J., Hose A.J. Consumption of Unprocessed Cow´s Milk Protects Infants from Common Respiratory Infections. Journal of Allergy Clinical Immunology, 2015, vol. 135, issue 1, pp. 56-62. https://doi.org/10.1016/j.jaci.2014.08.044Lupetti A., Paulusma-Annema A., Welling M. M., Dogterom-Ballering H., Brouwer C. P., Senesi S., Van Dissel J.P., Nibbering P.H. Synergistic activity of the n-terminal peptide of human lactoferrin and fluconazole against Candida species // Antimicrobial Agents and Chemotherapy. 2003. Vol. 47, issue 1. P. 262–267. https://doi.org/10.1128/aac.47.1.262–267.2003Lupetti A., Paulusma-Annema A., Welling M. M., Dogterom-Ballering H., Brouwer C. P., Senesi S., Van Dissel J.P., Nibbering P.H. Synergistic Activity of the N-Terminal Peptide of Human Lactoferrin and Fluconazole against Candida Species. Antimicrobial Agents and Chemotherapy, 2003, vol. 47, issue 1, pp. 262-267. https://doi.org/10.1128/aac.47.1.262-267.2003Paesano R., Pacifici E., Benedetti S., Berlutti F., Frioni A., Polimeni A., Valenti P. Safety and efficacy of Lactoferrin versus ferrous sulphate in curing Iron deficiency and iron deficiency anaemia in hereditary thrombophilia pregnant women: An interventional study // BioMetals. 2014. Vol. 27, issue 5. P. 999–1006. https://doi.org/10.1007/s10534-014-9723-xPaesano R., Pacifici E., Benedetti S., Berlutti F., Frioni A., Polimeni A., Valenti P. Safety and Efficacy of Lactoferrin Versus Ferrous Sulphate in Curing Iron Deficiency and Iron Deficiency Anaemia in Hereditary Thrombophilia Pregnant Women: An interventional Study. BioMetals, 2014, vol. 27, issue 5, pp. 999-1006. https://doi.org/10.1007/s10534-014-9723-xPanopoulos, G. Moatsou, C., Psychogyiopoulou C., Moschopoulou E. Microfiltration of ovine and bovine Milk: Effect on microbial counts and biochemical characteristics // Food. 2020. Vol. 9, issue 3. P. 284– 294. https://doi.org/10.3390/foods9030284Panopoulos, G. Moatsou, C., Psychogyiopoulou C., Moschopoulou, E. Microfiltration of Ovine and Bovine Milk: Effect on Microbial Counts and Biochemical Characteristics. Food, 2020, vol. 9, issue 3, pp. 284-294. https://doi.org/10.3390/foods9030284Pierce A., Legrand D. Lactoferrin: A Multifunctional Protein // Medecine Sciences. 2009. Vol. 25, issue 4. P. 361–369. https://doi.org/10.1051/medsci/2009254361Pierce A., Legrand D. Lactoferrin: A Multifunctional Protein, 2009, vol. 25, issue 4, pp. 361-369. https://doi.org/10.1051/medsci/2009254361Rashmi K., Warrier A. Non-thermal processing of dairy foods // Dairy Processing: Advanced Research to Applications / J. Minj, V. Sudhakaran, A. Kumari. Singapore: Springer, 2020. P. 25–49. https://doi.org/10.1007/978-981-15-2608-4_2Rashmi K., Warrier A. Non-thermal Processing of Dairy Foods. In J. Minj, V. Sudhakaran, A. Kumari. Dairy Processing: Advanced Research to Applications. Singapore: Springer, 2020. P. 25-49. https://doi.org/10.1007/978-981-15-2608-4_2Rynne N., Beresford T., Kelly A., Guinnee T. Effect of milk pasteurization temperature and in situ whey protein denaturation on the composition, texture and heat-induced functionality of half-fat cheddar cheese // International Dairy Journal. 2004. Vol. 14, issue 11. P. 989–1001. https://doi.org/10.1016/j.idairyj.2004.03.010Rynne N., Beresford T., Kelly A., Guinnee T. Effect of Milk Pasteurization Temperature and in Situ Whey Protein Denaturation on the Composition, Texture and Heat-Induced Functionality of HalfFat Cheddar Cheese. International Dairy Journal, 2004, vol. 14, issue 11, pp. 989-1001. https://doi.org/10.1016/j.idairyj.2004.03.010Sachdeva A., Nagpal J. Meta-analysis: efficacy of bovine lactoferrin in Helicobacter pylori eradication // Alimentary Pharmacology and Therapeutics. 2009. Vol. 29, issue 7. P. 720–730. https://doi.org/10.1111/j.1365–2036.2009.03934.xSachdeva A., Nagpal J. Meta‐analysis: efficacy of bovine lactoferrin in Helicobacter pylori eradication. Alimentary Pharmacology and Therapeutics, 2009, vol. 29, issue 7, pp. 720-730. https://doi.org/10.1111/j.1365-2036.2009.03934.xScarino M.L. A sideways glance: Take it or leave It? The role of Lactoferrin in Iron seguestration and delivery within the body // Genes & Nutrition. 2007. Vol. 2, issue 2. P. 161–162. https://doi.org/10.1007/s12263-007-0054-1Scarino M.L. A Sideways Glance: Take It or Leave It? The Role of Lactoferrin in Iron Seguestration and Delivery within the Body. Genes & Nutrition, 2007, vol. 2, issue 2, pp. 161-162. https://doi.org/10.1007/s12263-007-0054-1Tomasula P.M., Bonnaillie L.M. Crossflow microfiltration in the dairy industry // Emerging Dairy Processing Technologies: Opportunities for the Dairy Industry / N. Datta, P.M. Tomasula. N.Y.: John Wiley & SonsEditors, 2015. P. 1–32. https://doi.org/10.1002/9781118560471.ch1Tomasula P., Bonnaillie L. Crossflow Microfiltration in the Dairy Industry. In N. Datta, P.M. Tomasula. Emerging Dairy Processing Technologies: Opportunities for the Dairy Industry. N.Y.: John Wiley & SonsEditors, 2015. P. 1-32. https://doi.org/10.1002/9781118560471.ch1Wakabayashi H., Oda H., Yamauchi K., Abe F. Lactoferrin for prevention of common viral infections // Journal of Infection and Chemotherapy. 2014. Vol. 20, issue 11. P. 666–671. https://doi.org/10.1016/j.jiac.2014.08.003Wakabayashi H., Oda H., Yamauchi K., Abe F. Lactoferrin for Prevention of Common Viral Infections. Journal of Infection and Chemotherapy, 2014, vol. 20, issue 11, pp. 666-671. https://doi.org/10.1016/j.jiac.2014.08.003Ward P.P., Paz E., Conneely O.M. Multifunctional roles of Lactoferrin: A critical overview //Cellular and Molecular Life Sciences. 2005. Vol. 62. P. 2540–2548. https://doi.org/10.1007/s00018-005-5369-8Ward P.P., Paz E., Conneely O.M. Multifunctional Roles of Lactoferrin: A Critical Overview. Cellular and Molecular Life Sciences, 2005, vol. 62, pp. 2540-2548. https://doi.org/10.1007/s00018-005-5369-8

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