Isolation, purification, and characterization of staphylocoagulase, a blood coagulating protein from Staphylococcus sp. MBBJP S43
| Autor: | Jatin Kalita, Dhrubajyoti Das, Hari Prasanna Deka Boruah, B. G. Unni, Prasenjit Manna, Niren Kumar Dutta, Moonmee Bharadwaz |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Adult Coagulase Staphylococcus aureus Nitrogen Size-exclusion chromatography Biology medicine.disease_cause Biochemistry 03 medical and health sciences Structural Biology medicine Humans Molecular Biology Polyacrylamide gel electrophoresis Phylogeny Chromatography Temperature General Medicine Hydrogen-Ion Concentration biology.organism_classification Staphylocoagulase Enzyme assay Carbon 030104 developmental biology Sephadex biology.protein Staphylococcus Bacteria |
| Zdroj: | International journal of biological macromolecules. 102 |
| ISSN: | 1879-0003 |
| Popis: | Staphylocoagulase, a protein produced by S. aureus, play major role in blood coagulation and investigations are in advance to discover more staphylocoagulase producing species. The present study demonstrates the identification of a coagulase producing bacteria and isolation, purification and characterization of the protein. The bacteria was identified using 16S rDNA sequencing and phylogenetic investigation, classified the bacteria as Staphylococcus sp. MBBJP S43 with Genbank accession number KX907247. Tube test and Chromozym TH assay were used to study enzyme activity and comparison was made with five standard coagulase positive strains. The SEM images of the fibrin threads provide evidence of coagulation. The optimum temperature for enzyme activity was 37°C and pH of 6.5-7.5. Glucose and lactose as a carbon source and ammonium chloride as nitrogen source greatly influenced the bacterial growth. Staphylocoagulase has been purified to homogeneity (766 fold) by 80% (NH4)2SO4 precipitation, Sephadex G-75 gel filtration, DEAE anion exchange chromatography, and HPLC using C18 column. SDS PAGE revealed the molecular weight of the protein to be approximately 66kD and FTIR spectra of the purified protein demonstrated the presence of α helical structure. Present study revealed that the Staphylococcus sp. MBBJP S43 strain is a potential staphylocoagulase producing bacteria. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect