Purification and partial characterization of a low temperature responsive Mn-SOD from tea (Camellia sinensis (L.) O. Kuntze).
The manganese containing superoxide dismutase (Mn-SOD) was purified from a tea clone, TEENALI, which showed the lowest period of winter dormancy. Protein was purified using leaves of tea by ammonium sulfate precipitation, followed by column chromatography using DEAE-cellulose, and silica-based size exclusion chromatography on HPLC system. Upto 51-fold purification and a specific activity of 56.66 U/mg of protein was achieved, which yielded a single band upon denaturing PAGE. The enzyme had a native molecular weight of about 169 kDa, whereas a monomer with molecular weight of 43 kDa was found on SDS-PAGE suggesting it to be homotetramer. The purified enzyme had pH optima of 8.0. It exhibited a wide temperature range for its activity with optima at 0 degrees C suggesting its role in low temperature tolerance. The manuscript presents purification and characterization of high molecular weight Mn-SOD from tea and discusses its implication in tolerance of low temperature stress.
Vyas
Dhiraj
D
Biotechnology Division, Institute of Himalayan Bioresource Technology (CSIR), Palampur, Himachal Pradesh 176061, India. dhirajvyas@rediffmail.com
Kumar
Sanjay
S
D028241:Camellia sinensis / Q000201:enzymology* / Q000235:genetics; D002478:Cells, Cultured; D004789:Enzyme Activation; D004795:Enzyme Stability; D018869:Heat-Shock Response / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D018515:Plant Leaves / Q000201:enzymology* / Q000235:genetics; D013045:Species Specificity; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D013696:Temperature
D013482:Superoxide Dismutase