Abstract
In this study, a bacterium producing solvent-stable protease was isolated from salt-enriched soil after incubation in benzene and toluene enriched-media and identified as Lysinibacillus sphaericus PAP02 by 16S rRNA sequencing. Further, the enzyme was purified by 3.64-fold with yield of 12.07%. The purified protease molecular mass was determined to be 33 kDa by SDS-PAGE and the substrate activity was confirmed by the presence of a protease band. Further, the purified protease was characterized and found to be active at pH 10 and a temperature of 40°C. In addition, metal ions in the form of CoCl2 and ZnSO4 slightly upregulated the enzyme activity. The activity was reduced in EDTA, indicating that it was a metallo-type protease. The significant levels of protease were retained after incubation (14 days) in the presence of organic solvents at 25 and 50% concentrations. Activity was induced by hexane at both concentrations, but toluene only induced at 25%. The isolated organic solvent stable protease will be useful as a biocatalyst for many industrial applications, including peptide synthesis in non-aqueous media.
Acknowledgements
This study was supported by an Inha University Research Grant from Inha University, Republic of Korea.
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- Abbreviations
- DMF
dimethyl formamide
- DMSO
dimethyl sulfoxide
- EDTA
ethylenediaminetetraacetic acid
- PMSF
phenylmethylsulfonylfluoride
©2016 Institute of Molecular Biology, Slovak Academy of Sciences
Articles in the same Issue
- Synthesis of multifunctional γ-PGA-based superparamagnetic iron oxide nanoparticles for magnetic resonance imaging and controlled drug release
- Organic solvent stable protease isolation and characterization from organic solvent tolerant strain of Lysinibacillus sphaericus PAP02
- The impact of grazing absence in inland saline vegetation – a case study from Slovakia
- How dose of biochar and biochar with nitrogen can improve the parameters of soil organic matter and soil structure?
- Water repellent effects of manure amended soils on organic matter decomposition, C retention, and respired CO2-C
- Rainfall interception in a disturbed montane spruce (Picea abies) stand in the West Tatra Mountains
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- Meiobenthic Assemblages from the Southwestern Coast of the Black Sea, İğneada (Turkey)
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- Parasitic mesostigmatid mites (Acari) – common inhabitants of the nest boxes of starlings (Sturnus vulgaris) in a Polish urban habitat
- Mayfly (Insecta: Ephemeroptera) assemblages of a regulated perennial Mediterranean river system in the Western Balkans
- Distribution and ecology of the flightless bush-cricket Poecilimon schmidtii at its northern range margin
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Articles in the same Issue
- Synthesis of multifunctional γ-PGA-based superparamagnetic iron oxide nanoparticles for magnetic resonance imaging and controlled drug release
- Organic solvent stable protease isolation and characterization from organic solvent tolerant strain of Lysinibacillus sphaericus PAP02
- The impact of grazing absence in inland saline vegetation – a case study from Slovakia
- How dose of biochar and biochar with nitrogen can improve the parameters of soil organic matter and soil structure?
- Water repellent effects of manure amended soils on organic matter decomposition, C retention, and respired CO2-C
- Rainfall interception in a disturbed montane spruce (Picea abies) stand in the West Tatra Mountains
- ASL16 gene, a member of AS2/LOB family, is essential for lateral root formation in Arabidopsis
- Meiobenthic Assemblages from the Southwestern Coast of the Black Sea, İğneada (Turkey)
- Oviposition by selected water mite (Hydrachnidia) species from Lake Skadar and its catchment
- Parasitic mesostigmatid mites (Acari) – common inhabitants of the nest boxes of starlings (Sturnus vulgaris) in a Polish urban habitat
- Mayfly (Insecta: Ephemeroptera) assemblages of a regulated perennial Mediterranean river system in the Western Balkans
- Distribution and ecology of the flightless bush-cricket Poecilimon schmidtii at its northern range margin
- Sexual size monomorphism and body variation in the fat dormouse Glis glis in Slovakia