Isolation, characterisation and kinetic studies on SEVA TB ES-31 antigen, a metallo-serine protease of interest in serodiagnosis.

摘要: "Background: SEVA TB Excretory secretory -31 (ES-31) antigen, a glycoprotein isolated from M. tb H37Ra culture filtrate, was found to be useful in the serodiagnosis of pulmonary tuberculosis (TB), extrapulmonary and HIV-TB coinfection. Further, it has been shown zinc containing serine protease. Aim: To isolate purify ES-31 antigen H 37 Ra filtrate study its enzyme properties peptide sequence. Methods: purified strain by ammonium sulphate precipitation, SDS-PAGE fractionation FPLC. Protease activity studied using azocasein as substrate. further fractionated two dimensional polyacrylamide gel electrophoresis (2D PAGE) followed LCMS-T analysis. Results: Mycobacterial metallo-serine protease 3096 fold protein. Purified showed optimum at pH 7.0 0C. Of four substrates explored, maximum with had Km value 0.01 mM specific 6250 x 10 -6 U/mg analysis 2D PAGE protein spots (A B). Conclusion: Kinetic studies on protein, an immunogen metallo are reported for first time. structure biological role will interest."