The increasing size and density of the human population is leading to an increasing risk of infectious diseases that threaten to spread yet another pandemics. The widespread use of vaccination has reduced morbidity and mortality associated with viral infections and in some cases eradicated the virus from the population entirely. Regrettably, some virus species retain the ability to mutate rapidly and thus evade the vaccine-induced immune response. New antiviral drugs are therefore needed for the treatment and prevention of viral diseases. Modern research into the structures and properties of viral proteases, which are of key importance in the life cycle of viruses, makes it possible, in our opinion, to turn these enzymes into promising targets for the development of effective viral disease control methods.

Vozrastayushchiye s kazhdym godom chislennost' i plotnost' chelovecheskoy populyatsii privodit k uvelichivayushchemusya risku rasprostraneniya infektsionnykh zabolevaniy, chto grozit vozniknoveniyem vse novykh epidemiy po vsemu miru. Shirokoye ispol'zovaniye vaktsinatsii snizilo zabolevayemost' i smertnost', svyazannyye s virusnymi infektsiyami, a v nekotorykh sluchayakh polnost'yu unichtozhilo virus sredi naseleniya. K sozhaleniyu, nekotoryye vidy virusov sokhranyayut sposobnost' k bystroy mutatsii i takim obrazom uskol'zayut ot vyzvannogo vaktsinoy immunnogo otveta. V svyazi s etim dlya lecheniya i profilaktiki virusnykh zabolevaniy trebuyutsya novyye protivovirusnyye preparaty. Sovremennyye issledovaniya v oblasti struktur i svoystv virusnykh proteaz, imeyushchikh klyuchevoye znacheniye v zhiznennom tsikle virusov, pozvolyayut, na nash vzglyad, prevratit' eti fermenty v perspektivnyye misheni dlya razrabotki effektivnykh metodov bor'by s virusnymi zabolevaniyami.

The recently described bioluminescent system from fungi has great potential for developing highly efficient tools for biomedical research. Luciferase enzyme is one of the most crucial components of this system. The luciferase from Neonothopanus nambi fungus belongs to the novel still undescribed protein family. The structure data for this protein is almost absent. A detailed study of the N. nambi luciferase properties is necessary for the improvement of analytical methods based on the fungal bioluminescent system. Here we present the positions of key amino acid residues and their effect on enzyme function described using bioinformatic and experimental approaches. These results are useful for further fungal luciferase structure determination.

Morskie polihety Chaetopterus variopedatus (Renier) (semejstvo Chaetopteridae) — kosmopolity, predstavlyayushchie soboj vidovoj kompleks iz otdel'nyh populyacij-podvidov. Pri razdrazhenii chervi vypuskayut svetyashchiesya (460 nm) oblaka slizi, pri etom chasto yarko svetyatsya i ih parapodii. Na segodnyashnij den' po-prezhnemu ne yasno, kak imenno rabotaet biolyuminescentnaya sistema etih polihet. Ranee bylo vydvinuto predpolozhenie, chto lyuciferaza C. variopedatus mozhet byt' ispol'zovana dlya detekcii ferroptoza — nedavno otkrytogo puti programmiruemoj kletochnoj gibeli, vyzvannoj nakopleniem ionov dvuhvalentnogo zheleza. Cel'yu issledovaniya bylo vydelit' i oharakterizovat' lyuciferazy C. variopedatus, a takzhe sravnit' lyuciferazy C. variopedatus iz raznyh populyacij. Pri vydelenii otvetstvennogo za biolyuminescenciyu fermenta iz zamorozhennyh obrazcov brazil'skih C. variopedatus po usovershenstvovannoj metodike byli polucheny dve aktivnye lyuciferazy — L1 i L2. Predpolozhiv, chto odna iz ukazannyh lyuciferaz opredelyaet svechenie slizi, a drugaya — svechenie parapodij chervej, etu zhe metodiku primenili k razdel'nym obrazcam slizi i parapodij zhivyh dal'nevostochnyh C. variopedatus. Odnako ih sliz' okazalas' nesvetyashchejsya. Pokazano, chto funkciyu svecheniya parapodij polihet C. variopedatus obespechivaet lyuciferaza L2, tak kak ona obnaruzhena v obshchej biomasse brazil'skih polihet i v parapodiyah dal'nevostochnyh polihet. Svechenie slizi brazil'skih C. variopedatus obuslovleno funkcionirovaniem lyuciferazy L1, kotoraya otsutstvuet v slizi dal'nevostochnogo podvida. Nabor izoform lyuciferaz polihet C. variopedatus zavisit ot mesta ih obitaniya.

The marine polychaete Chaetopterus variopedatus (Renier) (family Chaetopteridae) is a cosmopolitan species complex, consisting of distinct populations/subspecies. The worms release glowing (460 nm) clouds of mucus when disturbed, and their parapodia often glow brightly. Currently, it is still unclear how exactly the bioluminescence system of these polychaetes functions. It has been previously assumed that the C. variopedatus luciferase may be used for detection of ferroptosis, the recently explored pathway of programmed cell death, resulting from accumulation of the ferrous ions. This study was aimed to extract and characterize the C. variopedatus luciferases, as well as to compare luciferases obtained from C. variopedatus of different populations. When extracting the enzyme responsible for bioluminescence from the frozen samples of Brazilian C. variopedatus using the improved method, two active luciferases, L1 and L2, were obtained. We assumed that one of the listed above luciferases was responsible for luminescence of the mucus and the other luciferase was responsible for luminescence in parapodia, and used the method for the distinct samples of mucus and parapodia of the living Far Eastern C. variopedatus. However, mucus of the latter turned out to be non-glowing. It is shown that luciferase L2 is responsible for luminescence in the parapodia of the C. variopedatus polychaete, since this luciferase has been found in the total biomass of Brazilian polychaetes and parapodia of Far Eastern polychaetes. Luminescence of the Brazilian C. variopedatus mucus is attributed to the functioning of luciferase L1, which is lacking in the mucus of the Far Eastern subspecies. The range of luciferase isoforms in polychaetes C. variopedatus depends on the place of origin.

A key component of the recently described bioluminescent system of higher fungi is luciferase, a new class of proteins. The properties of fungal luciferase and their relationship with its structure are interesting both for improving autoluminescent systems already created on its basis and for creating new ones. Therefore, it is extremely important to understand the spatial structure of this protein. We have performed heterologous expression and purification of Neonothopanus nambi luciferase, obtained a protein suitable for subsequent crystallization, and also determined some biochemical properties of the recombinant luciferase.

This paper presents the preliminary results of the separation of the Chaetopterus variopedatus bioluminescent system into luciferin and luciferase and a brief description of some of their properties.

Fouling of aquatic systems by harmful microalgal and cyanobacterial species is an environmental and public health concern. Microalgal bioreactors are engineered ecosystems for the cultivation of algal biomass to meet the increasing demand for alternative protein sources and algae-derived products. Such bioreactors are often open or semi-open ponds or raceways that are prone to contamination by contaminant photosynthetic microorganisms, including harmful cyanobacterial species (HCBs). HCBs affect the quality of products through the accumulation of off-flavours, reducing their acceptance by consumers, and through the production of several different toxins collectively known as cyanotoxins. The density of cultured species within the bioreactor environment creates difficulty in detecting low concentrations of contaminant cells, and there is currently no technology enabling rapid monitoring of contaminations. The present study demonstrates the potential of Low-Resolution Raman Spectroscopy (LRRS) as a tool for rapid detection of low concentrations of HCBs within dense populations of the spirulina (Arthrospira platensis) cultures. An LRRS system adapted for the direct measurement of raw biomass samples was used to assemble a database of Raman spectral signatures, from eight algal and cyanobacterial strains. This dataset was used to develop both quantitative and discriminative chemometric models. The results obtained from the chemometric analyses demonstrate the ability of the LRRS to detect and quantify algal and cyanobacterial species at concentrations as low as 103 cells/mL and to robustly discriminate between species at concentrations of 104 cells/mL. The LRRS and chemometric analyses were further able to detect the presence of low concentrations (103cells/mL) of contaminating species, including the toxic cyanobacterium Microcystis aeruginosa, within dense (>107 cells/mL) spirulina cultures. The results presented provide a first demonstration of the potential of LRRS technology for real-time detection of contaminant species within microalgal bioreactors, and possibly for early detection of developing harmful algal blooms in other aquatic ecosystems.

Представлены первые результаты разделения биолюминесцентной системы морской полихеты Chaetopterus variopedatus на люциферин и люциферазу, их очистки и краткого описания некоторых свойств.

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

By determining the components involved in the bioluminescence process in luminous and nonluminous organs of the honey fungus Armillaria mellea, we have established causes of partial luminescence of this fungus. The complete set of enzymes and substrates required for bioluminescence is formed only in the mycelium and only under the conditions of free oxygen access. Since the synthesis of luciferin precursor (hispidin) and 3-hydroxyhispidin hydroxylase in the fruiting bodies is blocked, the formation of luciferin-the key component of fungal bioluminescent system-was not observed. That is why the fruiting body of Armillaria mellea is nonluminous despite the presence of luciferase, the enzyme that catalyzes the oxidation of luciferin with a photon emission.

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.