Production of biotherapeutic drugs in mammalian cells, recombinant proteins in particular, may be handicapped by the limitations imposed on the cultures by metabolic burden. An alternative solution is to produce proteins in cells of other animals (e.g., Sf9, S2 and High Five insect cell lines, Caenorhabditis elegans and Schistosoma mansoni cell line) or orthogonal cell systems, including plant-based. In our opinion, non-traditional cell cultures may become promising tool for production of affordable and effective biotherapeutic drugs.

The deep learning technologies have become the driver of the revolutionary changes in scientific research in various fields. The AlphaFold-2 neural network software development that has solved the semicentennial problem of 3D protein structure prediction based on primary amino acid sequence is the most obvious example of using such technologies in structural biology and biomedicine. The use of deep learning methods for the prediction of protein–ligand interactions can considerably simplify predicting, speed up the development of new effective pharmaceuticals and change the concept of drug design.

Tekhnologii glubokogo obucheniya stali drajverom revolyucionnyh izmenenij v nauchnyh issledovaniyah raznyh oblastej. Naibolee yarkim primerom ih primeneniya v oblasti strukturnoj biologii i biomediciny yavlyaetsya programmnaya razrabotka nejroset' AlphaFold-2, reshivshaya poluvekovuyu problemu predskazaniya 3D-struktury belkov po pervichnoj aminokislotnoj posledovatel'nosti. Ispol'zovanie metodov glubokogo obucheniya dlya predskazaniya belok-ligandnyh vzaimodejstvij smozhet znachitel'no uprostit' predskazanie, uskorit' razrabotku novyh effektivnyh lekarstvennyh preparatov i pomenyat' koncepciyu drag-dizajna.

Proizvodstvo bioterapevticheskih preparatov, v chastnosti, rekombinantnyh belkov v kletkah mlekopitayushchih mozhet byt' zatrudneno iz-za ogranichenij ispol'zuemyh kul'tur v svyazi s metabolicheskoj nagruzkoj. Al'ternativnym podhodom dlya resheniya takih zadach yavlyaetsya narabotka belkov v kletkah drugih zhivotnyh (naprimer, kul'tury kletok nasekomyh Sf9, S2 i High Five, kul'tury kletok chervej vidov Caenorhabditis elegans i Schistosoma mansoni) ili ortogonal'nyh kletochnyh sistemah, v tom chisle rastitel'nyh. S nashej tochki zreniya, primenenie neklassicheskih kletochnyh kul'tur mozhet stat' perspektivnym napravleniem dlya polucheniya bolee dostupnyh i effektivnyh bioterapevticheskih preparatov.

Medicines play an indisputable role in life extension and improvement of the quality of life. To obtain medicinal compounds, researchers traditionally rely on natural sources and chemical synthesis, however, currently developing biotechnological methods allow one to introduce the group of genes encoding new metabolic pathways into the genomes of heterologous hosts and regulate activity of the hosts' intrinsic metabolic pathways. Such an approach makes it possible to reproduce biosynthesis of bioactive substances in heterologous hosts, the approach combines the benefits of conventional methods and works around the shorcomings of those. In our view, the use of metabolic engineering to obtain medicinal compounds is becoming increasingly important for their production.

Lekarstvennye preparaty igrayut neosporimuyu rol' v prodlenii zhizni i povyshenii ee kachestva. Dlya polucheniya lekarstvennyh soedinenij issledovateli tradicionno obrashchayutsya k prirodnym istochnikam i himicheskomu sintezu, odnako v nastoyashchee vremya aktivno razvivayutsya biotekhnologicheskie metody, pozvolyayushchie vnedryat' gruppy genov, kodiruyushchie novye metabolicheskie puti, v genomy geterologicheskih hozyaev i regulirovat' aktivnost' ih sobstvennyh metabolicheskih putej. Takoj podhod daet vozmozhnost' vosproizvodit' biosintez biologicheski aktivnyh soedinenij v geterologicheskih hozyaevah, sochetaet dostoinstva tradicionnyh metodov ih polucheniya i obhodit nedostatki etih metodov. S nashej tochki zreniya, primenenie metabolicheskoj inzhenerii dlya polucheniya lekarstvennyh soedinenij priobretaet vse bol'shee znachenie v proizvodstve.

Bioluminescence of dinoflagellates is a result of oxidation of the luciferin, which belongs to the class of tetrapyrroles and is possibly a product of chlorophyll a catabolism. We have demonstrated the functionality of the dinoflagellate luciferase gene from P. lunula in the transient and stable transformation of N. tabacum BY-2 cell culture. The results will be used further for the investigation of the luciferin biosynthetic pathway

Bioluminescent systems are increasingly being used for the development of highly sensitive optical imaging techniques in vivo. However, it is necessary to inject expensive and unstable synthetic substrates (luciferins) before each analysis for most of the systems applied. Autonomous bacterial and fungal bioluminescent systems, that recently have become available for implementation in eukaryotic cells, in our opinion, may be developed into an effective tool in new technologies of bioluminescent imaging.

Биолюминесцентные системы все чаще применяют для разработки высокочувствительных оптических методов имиджинга in vivo. Однако при использовании популярных систем необходимо инъекционно вводить дорогие и малостабильные синтетические субстраты (люциферины) перед каждым анализом. Автономные системы бактерий и грибов, которые недавно стали доступны для работы с эукариотическими клетками, по нашему мнению, могут развиться в полноценный инструмент для создания новых технологий биолюминесцентного имиджинга.

Bioluminescence is chemical oxidation of a small luciferin molecule by air catalyzed by luciferase and accompanied by the emission of photons in the visible spectrum. This reaction is used in bioluminescent bioimaging, the method for the visualization of organism’s interior. Bioimaging is a popular tool used in medical research. However, it has an unfortunate drawback: it requires introduction of external luciferin to the system before every experiment. In this work we discuss a possibility of developing an autonomous luminescent system in eukaryotes based on the bioluminescent system of higher fungi.

This is the first study to obtain a high-purity luciferase from the fungus Neonothopanus nambi biomass that is suitable for subsequent sequencing.

Современные биомедицинские исследования активно используют методы биоимиджинга клеток, тканей и целых организмов. Многоцветный биоимиджинг находит свое применение в случае необходимости одновременного наблюдения разных событий на молекулярном и клеточном уровнях. Наиболее чувствительными являются методы биолюминесцентного имиджинга, однако их использование для многоцветного мечения сдерживается недостаточным количеством доступных пар люцифераза–люциферин. Удачным расширением палитры инструментов молекулярного имиджинга могут стать новые биолюминесцентные системы высших грибов и морской полихеты Odontosyllis, обладающие рядом преимуществ по сравнению с ранее изученными системами.

Modern biomedical research technologies actively use bioimaging for studying cells, tissues and whole organisms. Multicolor bioimaging is applied when simultaneous observation of different events at the molecular and cellular level is needed. Bioluminescent imaging methods are the most sensitive, however, their use for multicolor labeling is complicated due to the insufficient number of available uciferin-luciferase pairs. Having a number of advantages compared to previously studied bioluminescent systems, the new bioluminescence systems of higher fungi and marine polychaete Odontosyllis could become a useful expansion of the bioimaging toolbox.

The structure of fungal oxyluciferin was determined, the enzymatic bioluminescence reaction under substrate saturation conditions with discrete monitoring of formed products was conducted, and the structures of the end products of the reaction were established. On the basis of these studies, the scheme of oxyluciferin degradation to the end products was developed. The structure of fungal oxyluciferin was confirmed by counter synthesis.

Определили структуру оксилюциферина грибов, провели ферментативную реакцию биолюминесценции в условиях насыщения по субстрату с дискретным мониторингом образующихся продуктов и установили структуры конечных продуктов реакции. На основе этих исследований разработали схему деградации оксилюциферина до конечных продуктов. Структуру оксилюциферина грибов подтвердили встречным синтезом.

New bioluminescent analogue of Fridericia luciferin was synthesized for the first time. Bioluminescence emission maximum of the compound demonstrates a 50-nm bathochromic shift compared to the luciferin. The obtained analogue may find use in the novel in vivo bioimaging applications.