RESUMO
Xylanase (EC 3. 2. 1. 8), hydrolyzes xylo-oligosaccharides into D-xylose and required for complete hydrolysis of native cellulose and biomass conversion. It has broad range of applications in the pulp and paper, pharmaceutical and Agri-food industries. Fifty fungal species were isolated from the fouled soil around an oil refinery and screened for the production of xylanase enzyme by enrichment culture techniques. The isolated fungal strain was identified as Hypocrea lixii SS1 based on the results of biochemical tests and 18s rRNA sequencing. The phylogenetic tree was constructed using the MEGA 5 software. Further, Hypocrea lixii SS1 was tested for the ability to utilize the sunflower oil sludge (waste from the oil industry) as the sole carbon source for xylanase production. The growth characteristics of Hypocrea lixii SS1 were also studied and maximum growth was found on the 7th day of incubation. The fungus showed a remarkable xylanase production of 38.9 U/mL. Xylanase was purified using a combination of 0-50% NH4SO2 precipitation, DEAE-sepharose and Sephacryl S-200 chromatography. Single peak obtained in RP-HPLC confirms the purity of xylanase. Further the enzyme produced was affirmed as xylanase with its molecular weight (29 kDa) using SDS-PAGE.
Assuntos
Microbiologia do Solo , Trichoderma/classificação , Trichoderma/isolamento & purificação , Xilosidases/análise , Cromatografia Líquida , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Eletroforese em Gel de Poliacrilamida , Dados de Sequência Molecular , Peso Molecular , Técnicas de Tipagem Micológica , Filogenia , /genética , Análise de Sequência de DNA , Trichoderma/enzimologia , Trichoderma/crescimento & desenvolvimento , Xilosidases/química , Xilosidases/isolamento & purificaçãoRESUMO
Xylanase (EC 3. 2. 1. 8), hydrolyzes xylo-oligosaccharides into D-xylose and required for complete hydrolysis of native cellulose and biomass conversion. It has broad range of applications in the pulp and paper, pharmaceutical and Agri-food industries. Fifty fungal species were isolated from the fouled soil around an oil refinery and screened for the production of xylanase enzyme by enrichment culture techniques. The isolated fungal strain was identified as Hypocrea lixii SS1 based on the results of biochemical tests and 18s rRNA sequencing. The phylogenetic tree was constructed using the MEGA 5 software. Further, Hypocrea lixii SS1 was tested for the ability to utilize the sunflower oil sludge (waste from the oil industry) as the sole carbon source for xylanase production. The growth characteristics of Hypocrea lixii SS1 were also studied and maximum growth was found on the 7th day of incubation. The fungus showed a remarkable xylanase production of 38.9 U/mL. Xylanase was purified using a combination of 0-50% NH4SO2 precipitation, DEAE-sepharose and Sephacryl S-200 chromatography. Single peak obtained in RP-HPLC confirms the purity of xylanase. Further the enzyme produced was affirmed as xylanase with its molecular weight (29 kDa) using SDS-PAGE.
Assuntos
Microbiologia do Solo , Trichoderma/classificação , Trichoderma/isolamento & purificação , Xilosidases/análise , Cromatografia Líquida , Análise por Conglomerados , DNA Fúngico/química , DNA Ribossômico/química , Eletroforese em Gel de Poliacrilamida , Dados de Sequência Molecular , Peso Molecular , Técnicas de Tipagem Micológica , FilogeniaRESUMO
Xylanase (EC 3. 2. 1. 8), hydrolyzes xylo-oligosaccharides into D-xylose and required for complete hydrolysis of native cellulose and biomass conversion. It has broad range of applications in the pulp and paper, pharmaceutical and Agri-food industries. Fifty fungal species were isolated from the fouled soil around an oil refinery and screened for the production of xylanase enzyme by enrichment culture techniques. The isolated fungal strain was identified as Hypocrea lixii SS1 based on the results of biochemical tests and 18s rRNA sequencing. The phylogenetic tree was constructed using the MEGA 5 software. Further, Hypocrea lixii SS1 was tested for the ability to utilize the sunflower oil sludge (waste from the oil industry) as the sole carbon source for xylanase production. The growth characteristics of Hypocrea lixii SS1 were also studied and maximum growth was found on the 7(th) day of incubation. The fungus showed a remarkable xylanase production of 38.9 U/mL. Xylanase was purified using a combination of 0-50% NH4SO2 precipitation, DEAE-sepharose and Sephacryl S-200 chromatography. Single peak obtained in RP-HPLC confirms the purity of xylanase. Further the enzyme produced was affirmed as xylanase with its molecular weight (29 kDa) using SDS-PAGE.
Assuntos
Microbiologia do Solo , Trichoderma/classificação , Trichoderma/isolamento & purificação , Xilosidases/análise , Cromatografia Líquida , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Eletroforese em Gel de Poliacrilamida , Dados de Sequência Molecular , Peso Molecular , Técnicas de Tipagem Micológica , Filogenia , RNA Ribossômico 18S/genética , Análise de Sequência de DNA , Trichoderma/enzimologia , Trichoderma/crescimento & desenvolvimento , Xilosidases/química , Xilosidases/isolamento & purificaçãoRESUMO
Degradation of the organophosphorus pesticide has been studied using the marine isolate, Streptomyces venezuelae ACT1. The organism exhibited a specific growth rate of 0.371h(-1) and the organophosphorus hydrolase activity rate as 0.273h(-1). Hence the organism was found to be very effective towards the pesticide degradation. Further the substrate assimilation and inhibition model of the organism were demonstrated using Monod and Haldane model equations which depicted that the inhibition model fits well for both the cell growth and enzyme activity. The maximum specific growth rate and the enzyme activity rate were found to be 0.571h(-1) and 0.472h(-1), respectively. Effect of P0/X0 ratio on degradation and COD reduction rate revealed that higher these ratios raise the degradation rate and the COD reduction rate.
Assuntos
Adaptação Biológica/fisiologia , Compostos Organofosforados/metabolismo , Resíduos de Praguicidas/metabolismo , Salinidade , Streptomyces/metabolismo , Poluentes Químicos da Água/metabolismo , Biodegradação Ambiental , Análise da Demanda Biológica de Oxigênio , Cinética , Modelos Químicos , Streptomyces/fisiologiaRESUMO
This investigation provides the enhanced production of thrombinase, a fibrinolytic enzyme using mutant Streptomyces venezuelae. Initially the mutagenesis of the marine isolate was done by UV and Ethyl methane sulfonate (EMS) and their mutational efficiencies were compared. The mutants were selected based on their high thrombinase activity and used for further studies. The mutant was found to be more halo and thermo tolerant comparing to wild. The effect of Dissolved oxygen level was also determined and the mutant offered the maximum specific growth rate as 0.2404 (h(-1)). The mutant showed high resistance to higher initial lactose concentration and the inhibition concentration was found to be 155.1mg/mL. The effect of S(0)/X(0) ratio on specific substrate consumption and production rate were also investigated. Both mutant and wild showed increase in specific substrate consumption and production rate at higher S(0)/X(0) ratio but the mutant showed better values than the wild strain.