Isolation, purification, and characterization of staphylocoagulase, a blood coagulating protein from Staphylococcus sp. MBBJP S43.

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% (NH)SO 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.