zldmЗаводская лаборатория. Диагностика материаловIndustrial laboratory. Diagnostics of materials1028-68612588-0187ООО «Издательство «ТЕСТ-ЗЛ»10.26896/1028-6861-2026-92-2-5-11zldm-2724Research ArticleАНАЛИЗ ВЕЩЕСТВАSUBSTANCES ANALYSISОпределение плесневого гриба Aspergillus niger амперометрическими тирозиназными биосенсорами, модифицированными наноматериалами, в биомедицинских объектах и сельскохозяйственной продукцииDetermination of Aspergillus niger fungus by amperometric tyrosinase biosensors modified with nanomaterials in biomedical objects and agricultural productsЛопаткоН. Ю.LopatkoN. Yu.

Надежда Юрьевна Лопатко

420008, г. Казань, Кремлевская ул., д. 18, корп. 1

Nadezhda Yu. Lopatko

18, bld. 1, Kremlevskaya ul., Kazan, 420008

БейлинсонР. М.BeilinsonR. M.

Регина Марковна Бейлинсон

420008, г. Казань, Кремлевская ул., д. 18, корп. 1

Regina M. Beilinson

18, bld. 1, Kremlevskaya ul., Kazan, 420008

rvarlamo@mail.ruМалоземоваО. Е.MalozemovaO. E.

Ольга Евгеньевна Малоземова

420008, г. Казань, Кремлевская ул., д. 18, корп. 1

Olga E. Malozemova

18, bld. 1, Kremlevskaya ul., Kazan, 420008

МедянцеваЭ. П.MedyantsevaE. P.

Эльвина Павловна Медянцева

420008, г. Казань, Кремлевская ул., д. 18, корп

Elvina P. Medyantseva

18, bld. 1, Kremlevskaya ul., Kazan, 420008

ХалдееваЕ. В.KhaldeevaE. V.

Елена Владимировна Халдеева

420015, г. Казань, Большая Красная ул., д. 67

Elena V. Khaldeeva

Laboratory of Mycology, 67, Bol’shaya Krasnaya ul., Kazan, 420015

УлаховичН. А.UlakhovichN. A.

Николай Алексеевич Улахович

420008, г. Казань, Кремлевская ул., д. 18, корп. 1

Nicolay A. Ulakhovich

18, bld. 1, Kremlevskaya ul., Kazan, 420008

Казанский (Приволжский) федеральный университетРоссияKazan (Volga Region) Federal UniversityRussian FederationКазанский НИИ эпидемиологии и микробиологии Роспотребнадзора, лаборатория микологииРоссияKazan Research Institute of Epidemiology and Microbiology of RospotrebnadzorRussian Federation202623022026922511Copyright © Лопатко Н.Ю., Бейлинсон Р.М., Малоземова О.Е., Медянцева Э.П., Халдеева Е.В., Улахович Н.А., 20262026Лопатко Н.Ю., Бейлинсон Р.М., Малоземова О.Е., Медянцева Э.П., Халдеева Е.В., Улахович Н.А.Lopatko N.Y., Beilinson R.M., Malozemova O.E., Medyantseva E.P., Khaldeeva E.V., Ulakhovich N.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.zldm.ru/jour/article/view/2724

Для определения антигена (Аг) плесневого гриба Aspergillus niger разработаны амперометрические биосенсоры на основе печатного графитового электрода, модифицированного многостенными углеродными нанотрубками (МУНТ), восстановленным оксидом графена (ВОГ) и наночастицами золота (НЧ Au). В качестве биочувствительного компонента использовали фермент тирозиназу. Установлено, что Аг Aspergillus niger является активатором тирозиназы в диапазоне содержаний 1 · 10–8 – 1 · 10–5 мкг/мг, нижняя граница определяемых содержаний cн = 7 · 10–9 мкг/мг. Модификация поверхности электрода углеродными наноматериалами и нанокомпозитом МУНТ/НЧ Au позволила улучшить ряд аналитических характеристик: расширить диапазон определения Аг Aspergillus niger до 5 · 10–10 – 1 · 10–3 мкг/мг в случае ВОГ, 1 · 10–10 – 1 · 10–3 мкг/мг в случае МУНТ и МУНТ/НЧ Au, снизить cн до 1 · 10–10 мкг/мг (модификатор ВОГ), 7 · 10–11 мкг/мг (МУНТ) и 5 · 10–11 мкг/мг (МУНТ/НЧ Au). Изучены кинетические параметры реакции ферментативного превращения фенола в присутствии Аг Aspergillus niger: независимо от наличия или отсутствия модификатора наблюдали двухпараметрическую согласованную активацию фермент-субстратной системы тирозиназа – фенол. Разработанные биосенсоры были апробированы для количественной оценки содержания Аг Aspergillus niger в образце репчатого лука и в образце бронхоальвеолярного лаважа (БАЛ). Правильность полученных результатов подтверждена альтернативными методами — микробиологическим (образец лука) и с использованием амперометрического иммуноферментного сенсора (образец БАЛ).

For determination of mold fungus antigen (Ag) Aspergillus niger, amperometric biosensors based on a printed graphite electrode modified with multi-walled carbon nanotubes (MWCNT), reduced graphene oxide (RGO), and gold nanoparticles (Au NP) were developed. Tyrosinase enzyme was used as a biosensitive component. It was found that Aspergillus niger Ag is an activator of tyrosinase in the concentration range of 1 × 10–8 – 1 × 10–5 μg/mg with LOD = 7 × 10–9 μg/mg. Modification of the electrode surface with carbon nanomaterials and the MWCNT/Au NPs nanocomposite allowed us to improve a number of analytical characteristics: expand the range of detectable concentrations to 1 × 10–10 – 1 × 10–3 μg/mg in the case of MWCNTs and MWCNTs/Au NPs, 5 × 10–10 – 1 × 10–3 μg/mg in the case of RGO, reduce the LOD to 7 × 10–11 μg/mg (MWCNT modifier), 1 × 10–10 μg/mg (RGO), 5 × 10–11 μg/mg (MWCNTs/Au NPs). Kinetic parameters of the reaction of enzymatic transformation of phenol in the presence of Ag Aspergillus niger were studied: regardless of the presence or absence of the modifier two-parameter coordinated activation of the enzyme-substrate tyrasinase – phenol system was observed. The developed biosensors were tested for the quantitative assessment of Aspergillus niger Ag content in a sample of agricultural crop (onion) and in a sample of bronchoalveolar lavage (BAL). The accuracy of the obtained results was confirmed by reference methods — microbiological (sample of onion) and using amperometric enzyme immunoassay sensor (BAL).

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