Purification of sarcosine oxidase enzyme by affinity chromatography

In this study, sarcosine bound cryogel columns have been developed and the efficiency in purification of monomeric sarcosine oxidase (MSOX) has been investigated. The sarcosine bound macroporous cryogels have been synthesized by free radical polymerization. For this purpose, metharcyloylamidohistidine (MAH) as the functional monomer and nano MAH-Sarcosine spheres have been synthesized and characterized by FT-IR in the first part of the study. Then, cryogel columns have been prepared by polymerization of MAH-Sarcosine and functional monomer in the presence of cross-linker N,N' Methylenebisacrylamide. Characterization of the prepared cryogel columns has been carried out by swelling tests, BET, SEM and FT-IR analysis. In the final step of the study, purification of MSOX from crude extracts of Bacillus cereus bacteria has been carried out by using prepared cryogel separation columns. Protein content of the purified MSOX has been determined by Bradford technique. Enzyme activity and kinetic studies have been carried out by UV spectrophotometric analysis. Molecular weight and purity degree of the purified enzyme have also been determined by SDS-PAGE analysis.