Salimeh Mohammadi, N. Hashim, N. Mahadi, Abdul Munir Abdul Murad
Glaciozyma antarctica is a psychrophilic yeast that was isolated from the surface of Antarctic sea ice. A key adaptation of psychrophilic microorganisms is to synthesize cold-active enzymes for survival at low temperatures. A full-length cDNA encoding β-glucanase (GaEgl) from G. antarctica PI12 was amplified by reverse-transcription polymerase chain reaction (RT-PCR). The cDNA encoded a 394-residue polypeptide with a putative signal peptide of 22 residues. Subsequently, the novel GaEgl was expressed in E. coli and purified with nickel affinity chromatography as an approximately 44 kDa protein. The biochemical characterisation of purified recombinant GaEgl (rGaEgl) revealed typical cold-active enzyme characteristics, such as maximal activity at 20 °C and pH 7.0. However, the enzyme was still active at 5-15 °C and alkaline pH values of 8-10. The activity of recombinant GaEgl was enhanced in the presence of Co2+ and Mn2+ metal ions. The Km and Vmax values of the enzyme using lichenan as the substrate were 8.87 mg mL-1 and 37.45 U mg-1, respectively. The enzymatic hydrolysis analysis of laminarin using HPLC showed that the main hydrolysis products were monosaccharides, disaccharides and trisaccharides. An analysis of the three-dimensional structure of the enzyme was carried out and compared with homologous mesophilic endo-β-1,3(4)-glucanase. The results of the comparative structural study revealed that the psychrophilic GaEgl contains longer loops, fewer hydrogen bonds and salt bridges, and a higher total solvent-accessible surface area which enhanced the protein flexibility for high catalytic efficiency at low temperatures.
{"title":"The Cold-Active Endo-β-1,3(4)-Glucanase from a Marine Psychrophilic Yeast, Glaciozyma antarctica PI12: Heterologous Expression, Biochemical Characterisation, and Molecular Modeling","authors":"Salimeh Mohammadi, N. Hashim, N. Mahadi, Abdul Munir Abdul Murad","doi":"10.26502/IJABPT.202101","DOIUrl":"https://doi.org/10.26502/IJABPT.202101","url":null,"abstract":"Glaciozyma antarctica is a psychrophilic yeast that was isolated from the surface of Antarctic sea ice. A key adaptation of psychrophilic microorganisms is to synthesize cold-active enzymes for survival at low temperatures. A full-length cDNA encoding β-glucanase (GaEgl) from G. antarctica PI12 was amplified by reverse-transcription polymerase chain reaction (RT-PCR). The cDNA encoded a 394-residue polypeptide with a putative signal peptide of 22 residues. Subsequently, the novel GaEgl was expressed in E. coli and purified with nickel affinity chromatography as an approximately 44 kDa protein. The biochemical characterisation of purified recombinant GaEgl (rGaEgl) revealed typical cold-active enzyme characteristics, such as maximal activity at 20 °C and pH 7.0. However, the enzyme was still active at 5-15 °C and alkaline pH values of 8-10. The activity of recombinant GaEgl was enhanced in the presence of Co2+ and Mn2+ metal ions. The Km and Vmax values of the enzyme using lichenan as the substrate were 8.87 mg mL-1 and 37.45 U mg-1, respectively. The enzymatic hydrolysis analysis of laminarin using HPLC showed that the main hydrolysis products were monosaccharides, disaccharides and trisaccharides. An analysis of the three-dimensional structure of the enzyme was carried out and compared with homologous mesophilic endo-β-1,3(4)-glucanase. The results of the comparative structural study revealed that the psychrophilic GaEgl contains longer loops, fewer hydrogen bonds and salt bridges, and a higher total solvent-accessible surface area which enhanced the protein flexibility for high catalytic efficiency at low temperatures.","PeriodicalId":13760,"journal":{"name":"International Journal of Applied Biology and P","volume":"11 1","pages":"279-300"},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86963555","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Safae El Mazouri, Tarik Aanniz, Jihane Touhtouh, Ilham Kandoussi, Mohammed Hakmi, L. Belyamani, A. Ibrahimi, Mouna Ouadghiri
The coronavirus disease 19 (Covid-19) pandemic caused by the SARS-CoV-2 virus has become a humanitarian crisis. Considering the severity of the situation we have performed a virtual screening of anthraquinones derivative drugs and phytochemicals targeting simultaneously multiple essential proteins of SARS-CoV-2 namely Mpro, PLpro, RdRp and the spike. Among the 9 screened anthraquinones derivative drugs, valrubicin, idarubicin, daunorubicin, doxorubicin, epirubicin and diacerein were the most potent inhibitors of SARS-CoV-2 Mpro, PLpro, RdRp and Spike simultaneously. Valrubicin has the best affinity towards the spike protein (-9.5 kcal/mol), RdRp (-8.2 kcal/mol) and PLpro (-7.9 kcal/mol) while idarubicin and doxorubicin were the most effective against Mpro (-8.3 kcal/mol). No toxicity measurements are required for these drugs since they were tested prior to their approval by the FDA. Of the 140 screened phytochemicals anthraquinones were the most potent candidates. Hypericin and rhein were able to bind to the active site of all four targets, while chrysophanol, aloesaponarin II, emodine, aloe-emodine, physcion and danthron simultaneously bound to the active site of SARS-CoV-2 Mpro, Spike and RdRp. Hypericin showed the best affinity towards the spike protein (-9.7 kcal/mol), RdRp (-10.2 kcal/mol) and PLpro (-7.8 kcal/mol), while chrysophanol was the most effective one against Mpro (-8.4 kcal/mol). Our overall prediction findings indicate that anthraquinones may inhibit the activity of the four essential proteins of SARS-CoV-2 and those results can pave the way in drug discovery.
{"title":"Anthraquinones: A Promising Multi-target Therapeutic Scaffold To Treat Covid-19","authors":"Safae El Mazouri, Tarik Aanniz, Jihane Touhtouh, Ilham Kandoussi, Mohammed Hakmi, L. Belyamani, A. Ibrahimi, Mouna Ouadghiri","doi":"10.26502/IJABPT.202104","DOIUrl":"https://doi.org/10.26502/IJABPT.202104","url":null,"abstract":"The coronavirus disease 19 (Covid-19) pandemic caused by the SARS-CoV-2 virus has become a humanitarian crisis. Considering the severity of the situation we have performed a virtual screening of anthraquinones derivative drugs and phytochemicals targeting simultaneously multiple essential proteins of SARS-CoV-2 namely Mpro, PLpro, RdRp and the spike. Among the 9 screened anthraquinones derivative drugs, valrubicin, idarubicin, daunorubicin, doxorubicin, epirubicin and diacerein were the most potent inhibitors of SARS-CoV-2 Mpro, PLpro, RdRp and Spike simultaneously. Valrubicin has the best affinity towards the spike protein (-9.5 kcal/mol), RdRp (-8.2 kcal/mol) and PLpro (-7.9 kcal/mol) while idarubicin and doxorubicin were the most effective against Mpro (-8.3 kcal/mol). No toxicity measurements are required for these drugs since they were tested prior to their approval by the FDA. Of the 140 screened phytochemicals anthraquinones were the most potent candidates. Hypericin and rhein were able to bind to the active site of all four targets, while chrysophanol, aloesaponarin II, emodine, aloe-emodine, physcion and danthron simultaneously bound to the active site of SARS-CoV-2 Mpro, Spike and RdRp. Hypericin showed the best affinity towards the spike protein (-9.7 kcal/mol), RdRp (-10.2 kcal/mol) and PLpro (-7.8 kcal/mol), while chrysophanol was the most effective one against Mpro (-8.4 kcal/mol). Our overall prediction findings indicate that anthraquinones may inhibit the activity of the four essential proteins of SARS-CoV-2 and those results can pave the way in drug discovery.","PeriodicalId":13760,"journal":{"name":"International Journal of Applied Biology and P","volume":"27 1","pages":"338-355"},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80306342","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
K. Ugoeze, Majiroghene Uyoyoumaero Okpa, N. Nwachukwu, B. Chinko, Kennedy Emeka Oluigbo
Abstract Background: Wounds are considered health indispositions with detrimental socio-economic influences on the afflicted and their kin. Curcuma longa or turmeric has been used in the treatment of wounds. Employing the extracts of turmeric as a bioactive ingredient in an aqueous cream could enhance its value in wound treatment. Aim: This study aimed to formulate aqueous creams containing concentrations of ethanolic extract of turmeric (EET) as bioactive ingredients, evaluate their stability and wound healing activities in male Wistar rats using hydroxyproline (HXP) as a biochemical marker. Materials and methods: Solutions of 1.5, 3.0 and 5.0 % w/v of crude EET were prepared and also incorporated as bioactive ingredients in aqueous creams. The stability of the creams was evaluated and their wound healing effects were studied using distilled water, dimethyl sulphoxide (DMSO) and cholesterol as controls in male Wistar rats. Results: The creams were stable in colour, pH, viscosity, etc. and exhibited wound healing activities. The animals treated with the crude 1.5 % w/v EET and its cream demonstrated the highest tissue HXP level showing significant percentage differences in tissue HXP levels from the control groups (p < 0.05). Conclusion: The crude EET has been confirmed to possess wound healing properties with an optimal effective concentration for wound healing in male Wistar rats determined as 1.5 % w/v which when incorporated as a bioactive ingredient in an aqueous cream retained its efficacy in wound healing and could therefore be beneficial in the treatment of body injuries.
{"title":"Evaluation of Aqueous Creams Containing Ethanolic Extract of Curcuma longa (Turmeric) as Bioactive Ingredient for the Management of Wounds","authors":"K. Ugoeze, Majiroghene Uyoyoumaero Okpa, N. Nwachukwu, B. Chinko, Kennedy Emeka Oluigbo","doi":"10.26502/IJABPT.202103","DOIUrl":"https://doi.org/10.26502/IJABPT.202103","url":null,"abstract":"Abstract Background: Wounds are considered health indispositions with detrimental socio-economic influences on the afflicted and their kin. Curcuma longa or turmeric has been used in the treatment of wounds. Employing the extracts of turmeric as a bioactive ingredient in an aqueous cream could enhance its value in wound treatment. Aim: This study aimed to formulate aqueous creams containing concentrations of ethanolic extract of turmeric (EET) as bioactive ingredients, evaluate their stability and wound healing activities in male Wistar rats using hydroxyproline (HXP) as a biochemical marker. Materials and methods: Solutions of 1.5, 3.0 and 5.0 % w/v of crude EET were prepared and also incorporated as bioactive ingredients in aqueous creams. The stability of the creams was evaluated and their wound healing effects were studied using distilled water, dimethyl sulphoxide (DMSO) and cholesterol as controls in male Wistar rats. Results: The creams were stable in colour, pH, viscosity, etc. and exhibited wound healing activities. The animals treated with the crude 1.5 % w/v EET and its cream demonstrated the highest tissue HXP level showing significant percentage differences in tissue HXP levels from the control groups (p < 0.05). Conclusion: The crude EET has been confirmed to possess wound healing properties with an optimal effective concentration for wound healing in male Wistar rats determined as 1.5 % w/v which when incorporated as a bioactive ingredient in an aqueous cream retained its efficacy in wound healing and could therefore be beneficial in the treatment of body injuries.","PeriodicalId":13760,"journal":{"name":"International Journal of Applied Biology and P","volume":"1 1","pages":"322-337"},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88561414","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abstract Corona pandemic triggered by SARS-CoV-2, the Severe Acute Respiratory Syndrome Corona Virus-2 has swept the world and created unique health-emergencies. Highly unrelated issues may have cumulated for a crisis of this magnitude to have occurred and can be aptly correlated to climatic and biodiversity changes that are deeply interlinked. It has emerged due to failing human health caused by climatic changes, over urbanization, and civilisations endangering the natural habitats on which they depend. As the pandemic progresses, a plethora of scientific and technical news keeps getting updated, causing a lot of anxieties on the therapeutics of corona virus and prior knowledge makes correlations among demography, climate and regional availabilities to manage the spread of the infection. This epidemic further altered the life style; causing huge losses and endangered the sustenance and livelihood of people. The review brings forth a consolidated story of the zoonotic origin of SARS-COV-2, its molecular structure, and discusses the ecological factors responsible for pandemic outbreak of Covid-19 and the strategies applied for the treatment. The list of “zoonotic diseases” includes bird flu, Ebola, HIV, Hendra, Zika, Hendra, SARS, and MERS. Having bats as its initial host, Corona then got communicated to humans directly or via a intermediate biological host. For any disease to become pandemic, ecological factors such as high population growth rate, malnutrition and unhygienic conditions, poor health sectors, environmental changes are responsible. Several environmental factors such as decrease in air-particulate matter, lowered emissions of CO2 and NO2 primarily because of lockdown leading to negligible human activities have caused reduction in air and water pollution in various countries. Undeniably, the worldwide air pollution is linked to industrialization, urbanization, consumption of excess fossil fuels etc. The unprecedented lockdown due to Covid-19 created a situation where the natural evolutionary processes protected and maintained the environmental concerns. This review deals with the necessity of controlling spread of disease in a sustainable way by nurturing nature and reminding the humans that this after all maybe the nature’s wake-up call?.
{"title":"Corona Outbreak: Is it Nature’s wake-up Call?","authors":"Kriti Bhardwaj Anita Kamra Verma Kapinder","doi":"10.26502/IJABPT.202102","DOIUrl":"https://doi.org/10.26502/IJABPT.202102","url":null,"abstract":"Abstract Corona pandemic triggered by SARS-CoV-2, the Severe Acute Respiratory Syndrome Corona Virus-2 has swept the world and created unique health-emergencies. Highly unrelated issues may have cumulated for a crisis of this magnitude to have occurred and can be aptly correlated to climatic and biodiversity changes that are deeply interlinked. It has emerged due to failing human health caused by climatic changes, over urbanization, and civilisations endangering the natural habitats on which they depend. As the pandemic progresses, a plethora of scientific and technical news keeps getting updated, causing a lot of anxieties on the therapeutics of corona virus and prior knowledge makes correlations among demography, climate and regional availabilities to manage the spread of the infection. This epidemic further altered the life style; causing huge losses and endangered the sustenance and livelihood of people. The review brings forth a consolidated story of the zoonotic origin of SARS-COV-2, its molecular structure, and discusses the ecological factors responsible for pandemic outbreak of Covid-19 and the strategies applied for the treatment. The list of “zoonotic diseases” includes bird flu, Ebola, HIV, Hendra, Zika, Hendra, SARS, and MERS. Having bats as its initial host, Corona then got communicated to humans directly or via a intermediate biological host. For any disease to become pandemic, ecological factors such as high population growth rate, malnutrition and unhygienic conditions, poor health sectors, environmental changes are responsible. Several environmental factors such as decrease in air-particulate matter, lowered emissions of CO2 and NO2 primarily because of lockdown leading to negligible human activities have caused reduction in air and water pollution in various countries. Undeniably, the worldwide air pollution is linked to industrialization, urbanization, consumption of excess fossil fuels etc. The unprecedented lockdown due to Covid-19 created a situation where the natural evolutionary processes protected and maintained the environmental concerns. This review deals with the necessity of controlling spread of disease in a sustainable way by nurturing nature and reminding the humans that this after all maybe the nature’s wake-up call?.","PeriodicalId":13760,"journal":{"name":"International Journal of Applied Biology and P","volume":"5 1","pages":"301-321"},"PeriodicalIF":0.0,"publicationDate":"2021-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81941262","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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