Purification and Characterization of a Thermostable Caseinolytic Serine Protease from the Latex of Euphorbia heterophylla L.

Title:Purification and Characterization of a Thermostable Caseinolytic Serine Protease from the Latex of Euphorbia heterophylla L.

Volume: 22 Issue: 9

Author(s): Sorokhaibam J. Singh, Laishram R. Singh, Sanjenbam K. Devi, Senjam S. Singh, Chingsubam B. Devi and Huidrom Rully

Affiliation:

Keywords: Euphorbia heterophylla, plant latex, protease, serine protease.

Abstract: A new thermostable caseinolytic serine protease was purified from the latex of Euphorbia heterophylla L. to electrophoretic homogeneity by a procedure involving successive steps of pretreatment of the latex, PEG fractionation, CM-cellulose chromatography and DEAE-cellulose chromatography. The purified protease was found to be a monomeric protein of molecular weight 77.2 kDa. It exhibited caseinolytic activity with hyperbolic azocasein saturation with V_max and K_m values of 0.11 units.mL^-1 and 0.55 mg.mL^-1 respectively. Specific inhibitory studies revealed the enzyme to be a serine protease. The protease was characterized by pH optimum of 8.0 and high thermostability with T_1/2 of 75°C. Based on the results of peptide mass fingerprinting analysis, the protease was shown to be a new protein not characterized earlier.

Cite this article as:

Singh J. Sorokhaibam, Singh R. Laishram, Devi K. Sanjenbam, Singh S. Senjam, Devi B. Chingsubam and Rully Huidrom, Purification and Characterization of a Thermostable Caseinolytic Serine Protease from the Latex of Euphorbia heterophylla L., Protein & Peptide Letters 2015; 22 (9) . https://dx.doi.org/10.2174/0929866522666150707114548