vuzbiochemiИзвестия вузов. Прикладная химия и биотехнологияProceedings of Universities. Applied Chemistry and Biotechnology2227-29252500-1558ИРНИТУ10.21285/2227-2925-2021-11-1-99-106vuzbiochemi-538Research ArticleФИЗИКО-ХИМИЧЕСКАЯ БИОЛОГИЯPHYSICOCHEMICAL BIOLOGYПолучение молочной кислоты из шелухи овсаObtaining lactic acid from oat husksШавыркинаН. А.ShavyrkinaN. A.

Шавыркина Надежда Александровна, к.т.н., доцент, старший научный сотрудник

659322, г. Бийск, ул. Социалистическая, 1

Nadezhda A. Shavyrkina, Cand. Sci. (Engineering), Associate Professor, Senior Scientist

1, Sotsialisticheskaya St., Biysk, 659322

32nadina@mail.ruСкибаE. А.SkibaE. A.

Скиба Екатерина Анатольевна, к.т.н., доцент, старший научный сотрудник

659322, г. Бийск, ул. Социалистическая, 1

Ekaterina A. Skiba, Cand. Sci. (Engineering), Associate Professor, Senior Scientist

1, Sotsialisticheskaya St., Biysk, 659322

eas08988@mail.ruИнститут проблем химико-энергетических технологий СО РАНРоссияInstitute for Problems of Chemical and Energetic Technologies SB RASRussian Federation20210504202111199106Copyright © Шавыркина Н.А., Скиба E.А., 20212021Шавыркина Н.А., Скиба E.А.Shavyrkina N.A., Skiba E.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vuzbiochemi.elpub.ru/jour/article/view/538

Полилактиды занимают лидирующую позицию в технологии получения биоразлагаемых полимеров для производства упаковочных материалов. В последнее время все чаще предлагается использовать дешевое целлюлозосодержащее сырье для получения прекурсора полилактидов – молочной кислоты. Авторами настоящей работы впервые в качестве сырья использована шелуха овса. Предварительная химическая обработка шелухи овса проведена в две стадии авторскими способами с использованием разбавленных растворов азотной кислоты и гидроксида натрия с получением технической целлюлозы. Далее техническая целлюлоза была подвергнута ферментативному гидролизу, после чего полученный преимущественно глюкозный ферментативный гидролизат сбраживался продуцентом Lactobacillus delbruеckii subsp. bulgaricus. Исследовано влияние на эффективность молочнокислого брожения активной кислотности среды, активизации закваски и дозировки дрожжевого экстракта. По результатам исследований рекомендуется поддерживать активную кислотность культуральной жидкости на уровне 6,5 ед. рН в течение всего процесса брожения, что позволяет повысить выход молочной кислоты в 1,7 раза по сравнению с процессом без корректировки рН. Показано, что активизация закваски путем пересева с молочной среды на глюкозную для переключения ферментативного аппарата продуцента с конверсии лактозы на конверсию глюкозы ускоряет процесс молочнокислого брожения в 2,2 раза и повышает константу сбраживания в 2,7 раза. Установлено, что внесение в среду дрожжевого экстракта в количестве 1% интенсифицирует молочнокислое брожение: процесс молочнокислого брожения ускоряется в 1,8 раза, константа сбраживания увеличивается в 1,4 раза, выход молочной кислоты повышается на 9,6% по сравнению с контролем и составляет 76,7%.

Polylactides are increasingly being used as biodegradable polymers in the production of packaging materials. It has been recently proposed to obtain a polylactide precursor – lactic acid – from inexpensive cellulose-containing raw materials. For the first time, the authors of the present work used oat husks for this purpose. Preliminary chemical treatment of oat husks was carried out in two stages using dilute solutions of nitric acid and sodium hydroxide to obtain technical cellulose, which was further subject to enzymatic hydrolysis. The as-obtained glucose enzymatic hydrolysate was fermented by the producer Lactobacillus delbrueckii subsp. bulgaricus. The effect of the active acidity of the medium, activation of the starter culture and dosage of the yeast extract on the efficiency of lactic acid fermentation was investigated. According to the obtained results, the active acidity of the culture should be maintained at a pH level of 6.5 during the entire fermentation process, which allows the yield of lactic acid to be increased by 1.7 times compared to the process without adjusting the pH. It was shown that the activation of the starter culture using a glucose medium instead of a milk medium, which switches the fermentation process from lactose conversion to glucose conversion, increases the rate of lactic acid fermentation by 2.2 times and the fermentation constant by 2.7 times. It was established that the introduction of yeast extract into the medium in an amount of 1% intensifies lactic acid fermentation: the process of lactic acid fermentation accelerates by 1.8 times; the fermentation constant increases by 1.4 times; the yield of lactic acid increases by 9.6% compared to the control thus reaching 76.7%.

шелуха овсамолочная кислотамолочнокислые бактерииполилактидыферментативный гидролизатoat huskslactic acidlactic acid bacteriapolylactidesenzymatic hydrolysateИсследование выполнено по бюджетному проекту ИПХЭТ СО РАН «Фундаментальные основы создания интегрированной технологии переработки легковозобновляемого непищевого растительного сырья в востребованные экономикой РФ продукты».The research was carried out according to the budget project of IPCET SB RAS "Fundamentals of the creation of an integrated technology for the processing of easily renewable non-food plant raw materials into products demanded by the Russian economy".ReferencesDe Oliveira R.A., Komesu A., Vaz Rossell C.E., Wolf Maciel M.R., Filho R.M. A study of the residual fermentation sugars influence on an alternative downstream process for first and second-generation lactic acid // Sustainable Chemistry and Pharmacy. 2020. Vol. 15. P. 100206. https://doi.org/10.1016/j.scp.2019.100206De Oliveira RA, Komesu A, Vaz Rossell CE, Wolf Maciel MR, Filho RM. A study of the residual fermentation sugars influence on an alternative downstream process for first and second-generation lactic acid. Sustainable Chemistry and Pharmacy. 2020;15:100206. https://doi.org/10.1016/j.scp.2019.100206Mazzoli R. Metabolic engineering strategies for consolidated production of lactic acid from lignocellulosic biomass // Biotechnology and Applied Biochemistry. 2020. Vol. 67. Issue 1. P. 61–72. https://doi.org/10.1002/bab.1869Mazzoli R. Metabolic engineering strategies for consolidated production of lactic acid from lignocellulosic biomass. Biotechnology and Applied Biochemistry. 2020;67(1):61–72. https://doi.org/10.1002/bab.1869Arfat Y.A., Ahmed J., Ejaz M., Mullah M. Polylactide/graphene oxide nanosheets/clove essential oil compositefilms for potential food packaging applications // International Journal of Biological Macromolecules. 2018. Vol. 107. Part A. P. 194–203. https://doi.org/10.1016/j.ijbiomac.2017.08.156Arfat YA, Ahmed J, Ejaz M, Mullah M. Polylactide/graphene oxide nanosheets/clove essential oil compositefilms for potential food packaging applications. International Journal of Biological Macromolecules. 2018;107:194–203. https://doi.org/10.1016/j.ijbiomac.2017.08.156Pagano N., Campana G., Fiorini М., Morelli R. Laser transmission welding of polylactide to aluminium thinfilms forapplications in the food-packaging industry // Optics & Laser Technology. 2017. Vol. 91. P. 80–84. https://doi.org/10.1016/j.optlastec.2016.12.014Pagano N, Campana G, Fiorini М, Morelli R. Laser transmission welding of polylactide to aluminium thinfilms forapplications in the food-packaging industry. Optics & Laser Technology. 2017;91:80–84. https://doi.org/10.1016/j.optlastec.2016.12.014Rapisarda M., Patanè C., Pellegrino A., Malvuccio A., Rizzo V., Muratore G., et al. Compostable Polylactide and Cellulose Based Packaging for Fresh-Cut Cherry Tomatoes: Performance Evaluation and Influence of Sterilization Treatment // Materials. 2020. Vol. 13. Issue 15. P. 3432. https://doi.org/10.3390/ma13153432Rapisarda M, Patanè C, Pellegrino A, Malvuccio A, Rizzo V, Muratore G, et al. Compostable Polylactide and Cellulose Based Packaging for Fresh-Cut Cherry Tomatoes: Performance Evaluation and Influence of Sterilization Treatment. Materials. 2020;13(15):3432. https://doi.org/10.3390/ma13153432Nashchekina Yu., Nikonov P., Nashchekin A., Mikhailova N. Functional Polylactide Blend Films for Controlling Mesenchymal Stem Cell Behaviour // Polymers. 2020. Vol. 12. Issue 9. P. 1969. https://doi.org/10.3390/polym12091969Nashchekina Yu, Nikonov P, Nashchekin A, Mikhailova N. Functional Polylactide Blend Films for Controlling Mesenchymal Stem Cell Behaviour. Polymers. 2020;12(9):1969. https://doi.org/10.3390/polym12091969Su Y., Nan G. Treatment of medial humeral epicondyle fractures in children using absorbable self-reinforced polylactide pins // Medicine. 2020. Vol. 99. Issue 17. P. e19861. https://doi.org/10.1097/MD.0000000000019861Su Y, Nan G. Treatment of medial humeral epicondyle fractures in children using absorbable self-reinforced polylactide pins. Medicine. 2020;99 (17):e19861. https://doi.org/10.1097/MD.0000000000019861Kolmakov A.G., Baikin A.S., Gudkov S.V., Belosludtsev K.N., Nasakina E.O., Kaplan M.A., et al. Polylactide-based stent coatings: biodegradable polymeric coatings capable of maintaining sustained release of the thrombolytic enzyme streptokinase // Pure and Applied Chemistry. 2020. Vol. 92. Issue 8. P. 1329–1340. https://doi.org/10.1515/pac-2019-1101Kolmakov AG, Baikin AS, Gudkov SV, Belosludtsev KN, Nasakina EO, Kaplan MA, et al. Polylactide-based stent coatings: biodegradable polymeric coatings capable of maintaining sustained release of the thrombolytic enzyme streptokinase. Pure and Applied Chemistry. 2020;92(8):1329–1340. https://doi.org/10.1515/pac-2019-1101Gherasim O., Grumezescu A.M., Grumezescu V., Iordache F., Vasile B.S., Holban A.M. Bioactive surfaces of polylactide and silver nanoparticles for the prevention of Microbial Contamination // Materials. 2020. Vol. 13. Issue 3. P. 768. https://doi.org/10.3390/ma13030768Gherasim O, Grumezescu AM, Grumezescu V, Iordache F, Vasile BS, Holban AM. Bioactive surfaces of polylactide and silver nanoparticles for the prevention of Microbial Contamination. Materials. 2020;13(3):768. https://doi.org/10.3390/ma13030768Zynov'eva M.E., Shnaider K.L., Zaripova S.K. Production of lactic acid on enzymatic hydrolysates of cellulose-containing raw materials // IOP Conference Series: Earth and Environmental Science. 2020. Vol. 421. P. 052011. https://doi.org/10.1088/1755-1315/421/5/052011Zynov'eva ME, Shnaider KL, Zaripova SK. Production of lactic acid on enzymatic hydrolysates of cellulose-containing raw materials. IOP Conference Series: Earth and Environmental Science. 2020;421:052011. https://doi.org/10.1088/1755-1315/421/5/052011Верхотурова Е.В., Лозовая Т.С., Евстафьев С.Н. Биосинтез молочной кислоты на основе продуктов автогидролиза соломы пшеницы // Известия вузов. Прикладная химия и биотехнология. 2016. Т. 6. N 3. P. 36–41. https://doi.org/10.21285/2227-2925-2016-6-3-36–41Verkhoturova EV, Lozovaya TS, Evstaf’ev SN. Biosynthesis of lactic acid via fermentation of wheat straw autohydrolysis products. Izvestiya Vuzov. Prikladnaya Khimiya i Biotekhnologiya = Proceedings of Universities. Applied Chemistry and Biotechnology. 2016; 6(3):36–41. (In Russian) https://doi.org/10.21285/22272925-2016-6-3-36–41Kobayashi K., et al. Method of producing chemical substance. Patent of United States, no. US 10,378,029 B2. 2019.Kobayashi K, et al. Method of producing chemical substance. Patent of United States, no. US 10,378,029 B2; 2019.Stephanopoulos G., et al. Method for producing lactic acid from organic waste. Patent of United States, no. US 10,174,347 B2. 2019.Stephanopoulos G, et al. Method for producing lactic acid from organic waste. Patent of United States, no. US 10,174,347 B2; 2019.Kashcheyeva E.I., Gismatulina Yu.A., Budaeva V.V. Pretreatments of non-woody cellulosic feedstocks for bacterial cellulose synthesis // Polymers. 2019. Vol. 11. Issue 10. P. 1645. https://doi.org/10.3390/polym11101645Kashcheyeva EI, Gismatulina YuA, Budaeva VV. Pretreatments of non-woody cellulosic feedstocks for bacterial cellulose synthesis. Polymers. 2019;11(10):1645. https://doi.org/10.3390/polym11101645Сакович Г.В., Будаева В.В., Скиба Е.А., Макарова Е.И., Павлов И.Н., Кортусов А.Н. [и др.]. Опыт масштабирования ферментативного гидролиза технических целлюлоз мискантуса и плодовых оболочек овса // Ползуновский вестник. 2012. N 4. С. 173–176.Sakovich GV, Budaeva VV, Skiba EA, Makarova EI, Pavlov IN, Kortusov AN, et al. Experience of scaling up the enzymatic hydrolysis of technical celluloses from miscanthus and oat hulls. Polzunovsky vestnik. 2012;4:173–176. (In Russian)Chopda R., Ferreira J.A., Taherzadeh M.J. Biorefining oat husks into high-quality lignin and enzymatically digestible cellulose with acid-catalyzed ethanol organosolv pretreatment // Processes. 2020. Vol. 8. Issue 4. P. 435. https://doi.org/10.3390/pr8040435Chopda R, Ferreira JA, Taherzadeh MJ. Biorefining oat husks into high-quality lignin and enzymatically digestible cellulose with acid-catalyzed ethanol organosolv pretreatment. Processes. 2020;8(4):435. https://doi.org/10.3390/pr8040435Santos J., Ouadi M., Jahangiri H., Hornung A. Thermochemical conversion of agricultural wastes applying different reforming temperatures // Fuel Processing Technology. 2020. Vol. 203. P. 106402. https://doi.org/10.1016/j.fuproc.2020.106402Santos J, Ouadi M, Jahangiri H, Hornung A. Thermochemical conversion of agricultural wastes applying different reforming temperatures. Fuel Processing Technology. 2020;203:106402. https://doi.org/10.1016/j.fuproc.2020.106402Sakovich G.V., Mikhailov Yu.M., Budaeva V.V., Korchagina A.A., Gismatulina Yu.A., Kozyrev N.V. Cellulose Nitrates from Unconventional Feedstocks // Doklady Chemistry. 2018. Vol. 483. Issue 1. P. 287– 291. https://doi.org/10.1134/S0012500818110101Sakovich GV, Mikhailov YuM, Budaeva VV, Korchagina AA, Gismatulina YuA, Kozyrev NV. Cellulose Nitrates from Unconventional Feedstocks. Doklady Chemistry. 2018;483(1):287–291. https://doi.org/10.1134/S0012500818110101Скиба Е.А., Будаева В.В., Макарова Е.И., Павлов И.Н., Золотухин В.Н., Сакович Г.В. Биоэтанол из целлюлозы плодовых оболочек овса // Вестник Казанского технологического университета. 2013. Т. 16. N 22. С. 202–205.Skiba EA, Budaeva VV, Makarova EI, Pavlov IN, Zolotukhin VN, Sakovich GV. Bioethanol from oat hulls cellulose. Vestnik Kazanskogo tekhnologicheskogo universiteta = Bulletin of Kazan Technological University. 2013;16(22):202–205 (In Russian)Гладышева Е.К., Скиба Е.А. Биосинтез бактериальной целлюлозы на ферментативном гидролизате технической целлюлозы из плодовых оболочек овса // Известия вузов. Прикладная химия и биотехнология. 2017. Т. 7. N 1. С. 141–147. https://doi.org/10.21285/2227-2925-2017-7-1-141-147Gladysheva EK, Skiba EA. Biosynthesis of bacterial cellulose on enzymatic hydrolyzate of oat hull pulp. Izvestiya Vuzov. Prikladnaya Khimiya i Biotekhnologiya = Proceedings of Universities. Applied Chemistry and Biotechnology. 2017;7(1):141–147. (In Russian) https://doi.org/10.21285/2227-2925-2017-7-1-141-147Технология спирта / В.Л. Яровенко, В.А. Маринченко, В.А. Смирнов [и др.]; под ред. проф. В.Л. Яровенко. М.: Колос-Пресс, 2002. 466 с.Yarovenko VL. (ed.) Alcohol technology. Moscow: Kolos-Press; 2002. 466 p. (In Russian).Karnaouri A., Asimakopoulou G., Kalogiannis K.G., Lappas A., Topakas E. Efficient D-lactic acid production by Lactobacillus delbrueckii subsp. bulgaricus through conversion of organosolv pretreated lignocellulosic biomass // Biomass and Bioenergy. 2020. Vol. 140. P. 105672. https://doi.org/10.1016/j.biombioe.2020.105672Karnaouri A, Asimakopoulou G, Kalogiannis KG, Lappas A, Topakas E. Efficient D-lactic acid production by Lactobacillus delbrueckii subsp. bulgaricus through conversion of organosolv pretreated lignocellulosic biomass. Biomass and Bioenergy. 2020;140:105672. https://doi.org/10.1016/j.biombioe.2020.105672

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