Proteases fromCanavalia ensiformis: Active and Thermostable Enzymes with Potential of Application in Biotechnology

Abstract

Extracts of leaves, seeds, roots, and stem from a tropical legume, C. ensiformis, were prepared employing buffers and detergent in aqueous solution. Leaf extracts had the highest protein content and the most pronounced peptidase activity with optimal pH in the neutral to alkaline range. All extracts exhibited peaks of activity at various pH values, suggesting the presence of distinctive classes of proteases. N-α-Tosyl-L-arginine methyl ester hydrolysis was maximal at 30°C to 60°C and peptidase activity from all extracts presented very good thermal stability after 24 h incubation at 70°C. C. ensiformis proteases exhibited molecular masses of about 200–57, 40–37, and 20–15 kDa by SDS-PAGE analysis. These enzymes cleaved hemoglobin, bovine serum albumin, casein, and gelatin at different levels. Serine and metalloproteases are the major proteases in C. ensiformis extracts, modulated by divalent cations, stable at 1% of surfactant Triton X-100 and at different concentrations of the reducing agent β-mercaptoethanol. Thus, C. ensiformis expresses a particular set of proteases in distinctive organs with high activity and stability, making this legume an important source of proteases with biotechnological potential.

Cajanus Cajan (l.) Millsp Aqueous Extracts against Melanoma Cell Line and their Proteases

“…The majority of plant extracts are prepared using organic solvents or a mixture of organic solvents with water to extract secondary metabolites that have hydrophobic characteristics. However, to extract proteins, which are polar molecules, has been employed water, buffers, or saline solutions [24,25]. As shown in Table 1, the C. cajan extracts presented different protein amounts.…”

“…This results indicated that water was a good protein extracting agent of stem, however, it was not so good for extracting proteins from roots, whereas, using similar amounts of starting material, the water extracted about twice more proteins from stem than from roots. Species of Fabaceae family have higher protein contents in the organs, such as Crotalaria spectabilis [24] and Canavalia ensiformis [25]. On the other hand, C. cajan has a lower protein content than the other legumes; however, high levels of proteins can be found in their seeds that are extensively studied [29].…”

Comparative Evaluation of Protease Extraction from Leaves of <i>Syzygium polyanthum</i>

“…Nevertheless, there are many types of extraction protocols used to extract protease from the plant leaves ranging from simple solvent-based extraction method to a more complex method which is associated with purification of the protease. For instance, simple extraction methods using distilled water and buffer solution are applied to extract protease from the plant leaves of Canavalia ensiformis where the leaves were first treated with liquid nitrogen to form powder [10]. In addition, the new serine protease namely populnein has also been extracted from the leaves of Thespesia populnea using sodium phosphate buffer solution followed by ammonium sulfate precipitation [6].…”

Debridement Applications of Bromelain: A Complex of Cysteine Proteases from Pineapple

“…They cleave peptide bounds in peptides or proteins and are found in all living organisms, in different parts of a cell and can catalyze in a great diversity of conditions. Therefore, there are many types of proteases in the nature, since they represent about 2% of protein coding genes and plants are excellent sources of proteolytic enzymes with great activity and stability [2,3].…”