Unpredictable influenza pandemics, annual epidemics, and sporadic poultry-to-human avian influenz... more Unpredictable influenza pandemics, annual epidemics, and sporadic poultry-to-human avian influenza virus infections with high morbidity and mortality rates dictate a need to develop new antiviral approaches. Targeting cellular pathways and processes is a promising antiviral strategy shown to be effective regardless of viral subtypes or viral evolution of drug-resistant variants. Proteomics-based searches provide a tool to reveal the druggable stages of the virus life cycle and to understand the putative antiviral mode of action of the drug(s). Ribonucleases (RNases) of different origins not only demonstrate antiviral effects that are mediated by the direct RNase action on viral and cellular RNAs but can also exert their impact by signal transduction modulation. To our knowledge, studies of the RNase-affected cell proteome have not yet been performed. To reveal cellular targets and explain the mechanisms underlying the antiviral effect employed by the small extra-cellular ribonucleas...
Bacillus species, generally regarded as soil microorganisms, are present in human gastrointestina... more Bacillus species, generally regarded as soil microorganisms, are present in human gastrointestinal tract (GIT) in quantities, which cannot be explained by their entrance with food only. They are capable of growing in GIT and interacting with intestinal microbiota and host organism by excretion of enzymes and low-molecular weight compounds, which exert digestion-facilitating, antagonistic, immunomodulating, antiviral, anticancer properties or mediate cell communication. For better understanding of its probiotic potential, we have sequenced genome of Bacillus altitudinis B-388 using mate-pair technology. It allowed us to improve quality of the genome sequence. The number of contigs decreased from 59 to 8. N50 contig length increased by four times. The number of identified genes raised from 3730 to 3774 (3645 proteins and 73 RNAs) with the reduction of frameshifted genes. The calculated size of B. altitudinis B-388 genome is 3,743,699 bp, with a G + C content of 41.17 mol%. Additional incomplete prophage sequence in genome of B. altitudinis B-388 was revealed. It was found that cryptic plasmid encodes SoxR, an oxidative stress response regulator. To date, the reported sequence is the most thorough presentation of B. altitudinis genome among four whole-genome sequences of this species deposited in GenBank.
Binase, a small guanyl-preferring extracellular ribonuclease of Gram-positive non-pathogenic soil... more Binase, a small guanyl-preferring extracellular ribonuclease of Gram-positive non-pathogenic soil bacteria Bacillus pumilus. Binase is a well-known bacterial ribonuclease, and the most essential properties of the enzyme were characterized. Binase has demonstrated antiviral activity in various virus-infected animal models. Most experiments associated with binase treatment of virus-infected animals were performed using single stranded RNA viruses. It is still unclear, whether binase is able to inactivate the double stranded RNA virus. Moreover, the phenomenon of the antiviral activity of binase against the DNA containing virus in animal model is not explained. Here, we presented the experimental results reflecting binase effect towards eukaryotic cells infected with viruses containing different types of nucleic acids. The obtained data revealed the bacterial ribonuclease binase mode of action against single stranded RNA influenza A virus, double stranded RNA reovirus and DNA containing herpes virus to prove future application of new antiviral tool with a broad range of activity.
Unpredictable influenza pandemics, annual epidemics, and sporadic poultry-to-human avian influenz... more Unpredictable influenza pandemics, annual epidemics, and sporadic poultry-to-human avian influenza virus infections with high morbidity and mortality rates dictate a need to develop new antiviral approaches. Targeting cellular pathways and processes is a promising antiviral strategy shown to be effective regardless of viral subtypes or viral evolution of drug-resistant variants. Proteomics-based searches provide a tool to reveal the druggable stages of the virus life cycle and to understand the putative antiviral mode of action of the drug(s). Ribonucleases (RNases) of different origins not only demonstrate antiviral effects that are mediated by the direct RNase action on viral and cellular RNAs but can also exert their impact by signal transduction modulation. To our knowledge, studies of the RNase-affected cell proteome have not yet been performed. To reveal cellular targets and explain the mechanisms underlying the antiviral effect employed by the small extra-cellular ribonucleas...
Reoviruses (respiratory enteric orphan viruses) are nonenveloped viruses with segmented dsRNA gen... more Reoviruses (respiratory enteric orphan viruses) are nonenveloped viruses with segmented dsRNA genome. Viruses in the family Reoviridae are quite stable in the environment. Recently, they have been identified with various pathologies and physiologic dysfunctions in a wide range of organs and tissues, including the hepatobiliary system, the myocardium, lungs, and endocrine tissues. Although most cases of reovirus infection are mild or subclinical diseases, the prevention measures are currently needed, especially for young children suffering from dehydrating gastroenteritis. To inhibit viral replication, different RNases targeting viral RNA are proposed. Here, we first have shown that RNase from Bacillus pumilus (binase) acts as an antiviral agent at the level of the whole animal organism infected by Mammalian orthoreovirus 1 strain Lang (TL1). The results obtained on the mice model infected with 10 LD50 and 20 LD50 doses of reovirus indicate the restoration of mice physiological parameters under binase treatment at the dose of 50 μg/mouse. Thus, our research supports the relevance of binase as a promising antiviral agent that affects viral RNA.
A Novel Antiviral Strategy against MERS-CoV and HCoV-229E Using Binase to Target Viral Genome Replication, 2017
RNA viruses cause most of the dangerous commu-nicable diseases. Due to their high mutation rates,... more RNA viruses cause most of the dangerous commu-nicable diseases. Due to their high mutation rates, RNA viruses quickly evade selective pressures and can adapt to a new host. Therefore, new antiviral approaches are urgently needed, which target more than one specific virus variant and which would optimally prevent development of viral resistance. Among the family of coronaviruses (CoV), several human pathogenic strains (HCoV) are known to cause respiratory diseases and are implied in enteric diseases. While most strains contribute to common cold-like illnesses, others lead to severe infections. One of these viruses is the newly emerged (2012), highly pathogenic Middle East respiratory syndrome coronavirus (MERS-CoV) of zoonotic origin. MERS-CoV causes a severe respiratory infection with a high mortality rate of 35 %. There is no specific treatment or infection prevention available. Here, we show that the bacterial ribonuclease Binase is able to inhibit the replication of MERS-CoV and of the low-pathogenic human coronavirus 229E (HCoV-229E) in cell culture. We demonstrate that at non-toxic concentrations, Binase decreased the titers of MERS-CoV and HCoV-229E. On a molecular level, Binase treatment reduced (i) the viral subgenomic RNAs and (ii) the viral nucleocapsidprotein (N) and non-structural protein 13 (nsp13) accumulation. Furthermore, we show that the quantity of the replication/transcription complexes within the infected cells is diminished. Thus, the data obtained might allow further development of new anti-coronaviral approaches affecting viral replication, independent of the specific virus strain.
Bacterial ribonuclease binase exerts an intra-cellular anti-viral mode of action targeting viral RNAs in influenza a virus-infected MDCK-II cells, Jan 5, 2018
Background: Influenza is a severe contagious disease especially in children, elderly and immunoco... more Background: Influenza is a severe contagious disease especially in children, elderly and immunocompromised patients. Beside vaccination, the discovery of new anti-viral agents represents an important strategy to encounter seasonal and pandemic influenza A virus (IAV) strains. The bacterial extra-cellular ribonuclease binase is a well-studied RNase from Bacillus pumilus. Treatment with binase was shown to improve survival of laboratory animals infected with different RNA viruses. Although binase reduced IAV titer in vitro and in vivo, the mode of action (MOA) of binase against IAV at the molecular level has yet not been studied in depth and remains elusive.
Многие рибонуклеазы (РНКазы) способны подавлять репродукцию РНК7содержащих вирусов в культурах ин... more Многие рибонуклеазы (РНКазы) способны подавлять репродукцию РНК7содержащих вирусов в культурах инфицированных клеток и лабораторных животных, однако молекулярные механизмы противовирусного действия РНКаз изучены недостаточно. В представленном обзоре рассмотрены наиболее изученные внутриклеточные РНКазы, обладающие противовирусной активностью (РНКаза L, белок MCPIP1, РНКазы эозинофилов), а также РНКазы, применяемые экзогенно (РНКаза А, BS7РНКаза, онконаза, биназа, синтетические РНКазы). Особое внимание уделено двум важным, хотя и не всегда обязательным свойствам этих РНКаз: каталитической активности и способности к димеризации. Предложена схема элиминации вируса экзогенными РНКазами, отражающая возможные типы взаимодействия РНКаз с вирусом и клеткой. Приведены доказательства того, что РНКазы, как классические компоненты иммунной защиты, представляют значительный интерес для разработки новых противовирусных препаратов.
Bacillus pumilus ribonuclease (binase) was shown to be a promising antiviral agent in animal mode... more Bacillus pumilus ribonuclease (binase) was shown to be a promising antiviral agent in animal models and cell cultures. However, the mode of its antiviral action remains unknown. To assess the binase effect on intracellular viral RNA we have selected single stranded negative-and positive-sense RNA viruses, influenza virus, and rhinovirus, respectively, which annually cause respiratory illnesses and are characterized by high contagious nature, mutation rate, and antigen variability. We have shown that binase exerts an antiviral effect on both viruses at the same concentration, which does not alter the spectrum of A549 cellular proteins and expression of housekeeping genes. The titers of influenza A (H1N1pdm) virus and human rhinovirus serotype 1A were reduced by 40% and 65%, respectively. A preincubation of influenza virus with binase before infection significantly reduced viral titer after single-cycle replication of the virus. Using influenza A virus mini genome system we showed that binase reduced GFP reporter signaling indicating a binase action on the expression of viral mRNA. Binase reduced the level of H1N1pdm viral NP mRNA accumulation in A549 cells by 20%. Since the viral mRNA is a possible target for binase this agent could be potentially applied in the antiviral therapy against both negative-and positive-sense RNA viruses.
Для исследования биосинтеза эндонуклеазы Serratia marcesacens в условиях блокирования биосинтеза ... more Для исследования биосинтеза эндонуклеазы Serratia marcesacens в условиях блокирования биосинтеза пуринов культуру выращивали в присутствии 0.1%-ного 2-(пара-аминобензолсульфамидо)-тиазола и по мере роста оценивали синтез эндонуклеазы по ее активности в питательной среде и периплазме бактерий. Показано, что при добавлении в питательную среду 2-(пара-аминобензолсульфамидо)-тиазола происходит качественное и количественное изменение динамики роста культуры S. marcescens W1050, увеличение биосинтеза и продукции эндонуклеазы, а также уменьшение доли внеклеточного и увеличение доли внутриклеточного фермента.
Сконструирована новая экспрессионная система на основе штамма B. megaterium PV 370. Ген новой риб... more Сконструирована новая экспрессионная система на основе штамма B. megaterium PV 370. Ген новой рибонуклеазы Bacillus sp.( RBT ) клонирован под контроль сильного Pspac промотора, транскрипция которого индуцируется изопропил- β - D -1-тиогалактопиранозидом (ИПТГ). Полученная векторная система позволит сократить время синтеза фермента и увеличить его выход в несколько раз в сравнении с природным продуцентом. Результаты проведенной работы могут служить основой для последующего выделения и очистки рекомбинантного белка RBT в препаративных количествах с целью исследования его биологических свойств.
В работе представлена полная методика выделения вирусов бактерий из разнообразных экологических н... more В работе представлена полная методика выделения вирусов бактерий из разнообразных экологических ниш Альметьевского и Высокогорского района Республики Татарстан (Россия). В качестве источников, содержащих бактериофаги, были использованы лесные и полевые (с. Борискино Альметьевского района), чернозёмные (с. Борискино, с. Куркачи Высокогорского района), городские нефтезагрязненные и городские чистые почвы (г. Альметьевск), а также вода сельского пруда (Борискино). В качестве индикаторных использованы 7 штаммов грамположительных и грамотрицательных бактерий. Получена коллекция из 24 фагов, специфически тропных к 7 штаммам индикаторных бактерий. Наибольшее количество фагов обнаружено у бактерий рода Bacillus.
The Journal of General and Applied Microbiology, 2016
The potential of microbial ribonucleases as promising antitumor and antiviral agents, determines ... more The potential of microbial ribonucleases as promising antitumor and antiviral agents, determines today's directions of their study. One direction is connected with biodiversity of RNases. We have analyzed completed and drafted Bacillus genomes deposited in GenBank for the presence of coding regions similar to the gene of an extracellular guanyl-preferring RNase of Bacillus amyloliquefaciens (barnase). Orthologues of the barnase gene were detected in 9 species out of 83. All of these belong to "B. subtilis" group within the genus. B. subtilis itself, as well as some other species within this group, lack such types of RNases. RNases similar to barnase were also found in species of "B. cereus" group as a part of plasmid-encoded S-layer toxins. It was also found that taxonomic states of culture collection strains, which were initially described based on a limited set of phenotypic characteristics, can be misleading and need to be confirmed. Using several approaches such as matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), sequencing of genes for 16S ribosomal RNA and RNA polymerase subunit beta followed by reconstruction of phylogenetic trees, we have re-identified two RNase-secreting Bacillus strains: B. thuringiensis B-388 which should be assigned as B. altitudinis B388 and B. intermedius 7P which should be renamed as B. pumilus 7P. Therefore, small secreted guanyl-preferring RNases are the feature of "B. subtilis" group only, which is characterized by distinctive lifestyle and adaptation strategies to environment.
Ribonucleases are considered as promising tools for anticancer treatment due to their selective c... more Ribonucleases are considered as promising tools for anticancer treatment due to their selective cytotoxicity against tumor cells. We investigated a new RNase from Bacillus altitudinis termed BALNASE (B. altitudinis RNase). Balnase is a close homolog of the well-known cytotoxic binase, differing by only one amino acid residue: nonpolar hydrophobic alanine at position 106 in the balnase molecule is replaced by a polar uncharged threonine in binase. The most exciting question is how the physico-chemical properties and biological effects of RNase might be changed by A106T substitution. Here, we have developed a chromatography-based rapid and modern technique for the purification of this new RNase which allowed us to get a protein sample of high quality with specific activity of 1.2 × 10(6) units in preparative amounts, suitable for further investigation of its biological properties.
Unpredictable influenza pandemics, annual epidemics, and sporadic poultry-to-human avian influenz... more Unpredictable influenza pandemics, annual epidemics, and sporadic poultry-to-human avian influenza virus infections with high morbidity and mortality rates dictate a need to develop new antiviral approaches. Targeting cellular pathways and processes is a promising antiviral strategy shown to be effective regardless of viral subtypes or viral evolution of drug-resistant variants. Proteomics-based searches provide a tool to reveal the druggable stages of the virus life cycle and to understand the putative antiviral mode of action of the drug(s). Ribonucleases (RNases) of different origins not only demonstrate antiviral effects that are mediated by the direct RNase action on viral and cellular RNAs but can also exert their impact by signal transduction modulation. To our knowledge, studies of the RNase-affected cell proteome have not yet been performed. To reveal cellular targets and explain the mechanisms underlying the antiviral effect employed by the small extra-cellular ribonucleas...
Bacillus species, generally regarded as soil microorganisms, are present in human gastrointestina... more Bacillus species, generally regarded as soil microorganisms, are present in human gastrointestinal tract (GIT) in quantities, which cannot be explained by their entrance with food only. They are capable of growing in GIT and interacting with intestinal microbiota and host organism by excretion of enzymes and low-molecular weight compounds, which exert digestion-facilitating, antagonistic, immunomodulating, antiviral, anticancer properties or mediate cell communication. For better understanding of its probiotic potential, we have sequenced genome of Bacillus altitudinis B-388 using mate-pair technology. It allowed us to improve quality of the genome sequence. The number of contigs decreased from 59 to 8. N50 contig length increased by four times. The number of identified genes raised from 3730 to 3774 (3645 proteins and 73 RNAs) with the reduction of frameshifted genes. The calculated size of B. altitudinis B-388 genome is 3,743,699 bp, with a G + C content of 41.17 mol%. Additional incomplete prophage sequence in genome of B. altitudinis B-388 was revealed. It was found that cryptic plasmid encodes SoxR, an oxidative stress response regulator. To date, the reported sequence is the most thorough presentation of B. altitudinis genome among four whole-genome sequences of this species deposited in GenBank.
Binase, a small guanyl-preferring extracellular ribonuclease of Gram-positive non-pathogenic soil... more Binase, a small guanyl-preferring extracellular ribonuclease of Gram-positive non-pathogenic soil bacteria Bacillus pumilus. Binase is a well-known bacterial ribonuclease, and the most essential properties of the enzyme were characterized. Binase has demonstrated antiviral activity in various virus-infected animal models. Most experiments associated with binase treatment of virus-infected animals were performed using single stranded RNA viruses. It is still unclear, whether binase is able to inactivate the double stranded RNA virus. Moreover, the phenomenon of the antiviral activity of binase against the DNA containing virus in animal model is not explained. Here, we presented the experimental results reflecting binase effect towards eukaryotic cells infected with viruses containing different types of nucleic acids. The obtained data revealed the bacterial ribonuclease binase mode of action against single stranded RNA influenza A virus, double stranded RNA reovirus and DNA containing herpes virus to prove future application of new antiviral tool with a broad range of activity.
Unpredictable influenza pandemics, annual epidemics, and sporadic poultry-to-human avian influenz... more Unpredictable influenza pandemics, annual epidemics, and sporadic poultry-to-human avian influenza virus infections with high morbidity and mortality rates dictate a need to develop new antiviral approaches. Targeting cellular pathways and processes is a promising antiviral strategy shown to be effective regardless of viral subtypes or viral evolution of drug-resistant variants. Proteomics-based searches provide a tool to reveal the druggable stages of the virus life cycle and to understand the putative antiviral mode of action of the drug(s). Ribonucleases (RNases) of different origins not only demonstrate antiviral effects that are mediated by the direct RNase action on viral and cellular RNAs but can also exert their impact by signal transduction modulation. To our knowledge, studies of the RNase-affected cell proteome have not yet been performed. To reveal cellular targets and explain the mechanisms underlying the antiviral effect employed by the small extra-cellular ribonucleas...
Reoviruses (respiratory enteric orphan viruses) are nonenveloped viruses with segmented dsRNA gen... more Reoviruses (respiratory enteric orphan viruses) are nonenveloped viruses with segmented dsRNA genome. Viruses in the family Reoviridae are quite stable in the environment. Recently, they have been identified with various pathologies and physiologic dysfunctions in a wide range of organs and tissues, including the hepatobiliary system, the myocardium, lungs, and endocrine tissues. Although most cases of reovirus infection are mild or subclinical diseases, the prevention measures are currently needed, especially for young children suffering from dehydrating gastroenteritis. To inhibit viral replication, different RNases targeting viral RNA are proposed. Here, we first have shown that RNase from Bacillus pumilus (binase) acts as an antiviral agent at the level of the whole animal organism infected by Mammalian orthoreovirus 1 strain Lang (TL1). The results obtained on the mice model infected with 10 LD50 and 20 LD50 doses of reovirus indicate the restoration of mice physiological parameters under binase treatment at the dose of 50 μg/mouse. Thus, our research supports the relevance of binase as a promising antiviral agent that affects viral RNA.
A Novel Antiviral Strategy against MERS-CoV and HCoV-229E Using Binase to Target Viral Genome Replication, 2017
RNA viruses cause most of the dangerous commu-nicable diseases. Due to their high mutation rates,... more RNA viruses cause most of the dangerous commu-nicable diseases. Due to their high mutation rates, RNA viruses quickly evade selective pressures and can adapt to a new host. Therefore, new antiviral approaches are urgently needed, which target more than one specific virus variant and which would optimally prevent development of viral resistance. Among the family of coronaviruses (CoV), several human pathogenic strains (HCoV) are known to cause respiratory diseases and are implied in enteric diseases. While most strains contribute to common cold-like illnesses, others lead to severe infections. One of these viruses is the newly emerged (2012), highly pathogenic Middle East respiratory syndrome coronavirus (MERS-CoV) of zoonotic origin. MERS-CoV causes a severe respiratory infection with a high mortality rate of 35 %. There is no specific treatment or infection prevention available. Here, we show that the bacterial ribonuclease Binase is able to inhibit the replication of MERS-CoV and of the low-pathogenic human coronavirus 229E (HCoV-229E) in cell culture. We demonstrate that at non-toxic concentrations, Binase decreased the titers of MERS-CoV and HCoV-229E. On a molecular level, Binase treatment reduced (i) the viral subgenomic RNAs and (ii) the viral nucleocapsidprotein (N) and non-structural protein 13 (nsp13) accumulation. Furthermore, we show that the quantity of the replication/transcription complexes within the infected cells is diminished. Thus, the data obtained might allow further development of new anti-coronaviral approaches affecting viral replication, independent of the specific virus strain.
Bacterial ribonuclease binase exerts an intra-cellular anti-viral mode of action targeting viral RNAs in influenza a virus-infected MDCK-II cells, Jan 5, 2018
Background: Influenza is a severe contagious disease especially in children, elderly and immunoco... more Background: Influenza is a severe contagious disease especially in children, elderly and immunocompromised patients. Beside vaccination, the discovery of new anti-viral agents represents an important strategy to encounter seasonal and pandemic influenza A virus (IAV) strains. The bacterial extra-cellular ribonuclease binase is a well-studied RNase from Bacillus pumilus. Treatment with binase was shown to improve survival of laboratory animals infected with different RNA viruses. Although binase reduced IAV titer in vitro and in vivo, the mode of action (MOA) of binase against IAV at the molecular level has yet not been studied in depth and remains elusive.
Многие рибонуклеазы (РНКазы) способны подавлять репродукцию РНК7содержащих вирусов в культурах ин... more Многие рибонуклеазы (РНКазы) способны подавлять репродукцию РНК7содержащих вирусов в культурах инфицированных клеток и лабораторных животных, однако молекулярные механизмы противовирусного действия РНКаз изучены недостаточно. В представленном обзоре рассмотрены наиболее изученные внутриклеточные РНКазы, обладающие противовирусной активностью (РНКаза L, белок MCPIP1, РНКазы эозинофилов), а также РНКазы, применяемые экзогенно (РНКаза А, BS7РНКаза, онконаза, биназа, синтетические РНКазы). Особое внимание уделено двум важным, хотя и не всегда обязательным свойствам этих РНКаз: каталитической активности и способности к димеризации. Предложена схема элиминации вируса экзогенными РНКазами, отражающая возможные типы взаимодействия РНКаз с вирусом и клеткой. Приведены доказательства того, что РНКазы, как классические компоненты иммунной защиты, представляют значительный интерес для разработки новых противовирусных препаратов.
Bacillus pumilus ribonuclease (binase) was shown to be a promising antiviral agent in animal mode... more Bacillus pumilus ribonuclease (binase) was shown to be a promising antiviral agent in animal models and cell cultures. However, the mode of its antiviral action remains unknown. To assess the binase effect on intracellular viral RNA we have selected single stranded negative-and positive-sense RNA viruses, influenza virus, and rhinovirus, respectively, which annually cause respiratory illnesses and are characterized by high contagious nature, mutation rate, and antigen variability. We have shown that binase exerts an antiviral effect on both viruses at the same concentration, which does not alter the spectrum of A549 cellular proteins and expression of housekeeping genes. The titers of influenza A (H1N1pdm) virus and human rhinovirus serotype 1A were reduced by 40% and 65%, respectively. A preincubation of influenza virus with binase before infection significantly reduced viral titer after single-cycle replication of the virus. Using influenza A virus mini genome system we showed that binase reduced GFP reporter signaling indicating a binase action on the expression of viral mRNA. Binase reduced the level of H1N1pdm viral NP mRNA accumulation in A549 cells by 20%. Since the viral mRNA is a possible target for binase this agent could be potentially applied in the antiviral therapy against both negative-and positive-sense RNA viruses.
Для исследования биосинтеза эндонуклеазы Serratia marcesacens в условиях блокирования биосинтеза ... more Для исследования биосинтеза эндонуклеазы Serratia marcesacens в условиях блокирования биосинтеза пуринов культуру выращивали в присутствии 0.1%-ного 2-(пара-аминобензолсульфамидо)-тиазола и по мере роста оценивали синтез эндонуклеазы по ее активности в питательной среде и периплазме бактерий. Показано, что при добавлении в питательную среду 2-(пара-аминобензолсульфамидо)-тиазола происходит качественное и количественное изменение динамики роста культуры S. marcescens W1050, увеличение биосинтеза и продукции эндонуклеазы, а также уменьшение доли внеклеточного и увеличение доли внутриклеточного фермента.
Сконструирована новая экспрессионная система на основе штамма B. megaterium PV 370. Ген новой риб... more Сконструирована новая экспрессионная система на основе штамма B. megaterium PV 370. Ген новой рибонуклеазы Bacillus sp.( RBT ) клонирован под контроль сильного Pspac промотора, транскрипция которого индуцируется изопропил- β - D -1-тиогалактопиранозидом (ИПТГ). Полученная векторная система позволит сократить время синтеза фермента и увеличить его выход в несколько раз в сравнении с природным продуцентом. Результаты проведенной работы могут служить основой для последующего выделения и очистки рекомбинантного белка RBT в препаративных количествах с целью исследования его биологических свойств.
В работе представлена полная методика выделения вирусов бактерий из разнообразных экологических н... more В работе представлена полная методика выделения вирусов бактерий из разнообразных экологических ниш Альметьевского и Высокогорского района Республики Татарстан (Россия). В качестве источников, содержащих бактериофаги, были использованы лесные и полевые (с. Борискино Альметьевского района), чернозёмные (с. Борискино, с. Куркачи Высокогорского района), городские нефтезагрязненные и городские чистые почвы (г. Альметьевск), а также вода сельского пруда (Борискино). В качестве индикаторных использованы 7 штаммов грамположительных и грамотрицательных бактерий. Получена коллекция из 24 фагов, специфически тропных к 7 штаммам индикаторных бактерий. Наибольшее количество фагов обнаружено у бактерий рода Bacillus.
The Journal of General and Applied Microbiology, 2016
The potential of microbial ribonucleases as promising antitumor and antiviral agents, determines ... more The potential of microbial ribonucleases as promising antitumor and antiviral agents, determines today's directions of their study. One direction is connected with biodiversity of RNases. We have analyzed completed and drafted Bacillus genomes deposited in GenBank for the presence of coding regions similar to the gene of an extracellular guanyl-preferring RNase of Bacillus amyloliquefaciens (barnase). Orthologues of the barnase gene were detected in 9 species out of 83. All of these belong to "B. subtilis" group within the genus. B. subtilis itself, as well as some other species within this group, lack such types of RNases. RNases similar to barnase were also found in species of "B. cereus" group as a part of plasmid-encoded S-layer toxins. It was also found that taxonomic states of culture collection strains, which were initially described based on a limited set of phenotypic characteristics, can be misleading and need to be confirmed. Using several approaches such as matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), sequencing of genes for 16S ribosomal RNA and RNA polymerase subunit beta followed by reconstruction of phylogenetic trees, we have re-identified two RNase-secreting Bacillus strains: B. thuringiensis B-388 which should be assigned as B. altitudinis B388 and B. intermedius 7P which should be renamed as B. pumilus 7P. Therefore, small secreted guanyl-preferring RNases are the feature of "B. subtilis" group only, which is characterized by distinctive lifestyle and adaptation strategies to environment.
Ribonucleases are considered as promising tools for anticancer treatment due to their selective c... more Ribonucleases are considered as promising tools for anticancer treatment due to their selective cytotoxicity against tumor cells. We investigated a new RNase from Bacillus altitudinis termed BALNASE (B. altitudinis RNase). Balnase is a close homolog of the well-known cytotoxic binase, differing by only one amino acid residue: nonpolar hydrophobic alanine at position 106 in the balnase molecule is replaced by a polar uncharged threonine in binase. The most exciting question is how the physico-chemical properties and biological effects of RNase might be changed by A106T substitution. Here, we have developed a chromatography-based rapid and modern technique for the purification of this new RNase which allowed us to get a protein sample of high quality with specific activity of 1.2 × 10(6) units in preparative amounts, suitable for further investigation of its biological properties.
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онконаза, биназа, синтетические РНКазы). Особое внимание уделено двум важным, хотя и не всегда обязательным свойствам этих РНКаз: каталитической активности и способности к димеризации. Предложена схема элиминации вируса экзогенными РНКазами, отражающая возможные типы взаимодействия РНКаз с вирусом и клеткой. Приведены доказательства того, что РНКазы, как классические компоненты иммунной защиты, представляют значительный интерес для разработки новых противовирусных препаратов.
онконаза, биназа, синтетические РНКазы). Особое внимание уделено двум важным, хотя и не всегда обязательным свойствам этих РНКаз: каталитической активности и способности к димеризации. Предложена схема элиминации вируса экзогенными РНКазами, отражающая возможные типы взаимодействия РНКаз с вирусом и клеткой. Приведены доказательства того, что РНКазы, как классические компоненты иммунной защиты, представляют значительный интерес для разработки новых противовирусных препаратов.