Selective and reversible thiol-pegylation, an effective approach for purification and characterization of five fully active ficin (iso)forms from Ficus carica latex

A novel form of ficin from Ficus carica latex: Purification and characterization

“…2), clearly indicated that ficin E was mainly eluted in peak b, with however some presence in peak a. eluted peaks were devoid of amidase activity and were thus discarded. As shown by N-terminal sequencing, peak c contained exclusively ficin B, eluting after ficin A (Azarkan et al, 2011).…”

“…As the five ficin (iso)forms already published were characterized with apparent molecular weights of $24 kDa, we focused our attention on the 27 kDa molecular weight band attributed to a new ficin form, named ficin E. Given the small difference of $3 kDa in molecular weights for the ficins contained in pool I (ficin A in Azarkan et al, 2011) we could not try a separation, e.g., by gel filtration chromatography. Therefore we submitted this protein pool (ficins A/E) to hydrophobic interaction chromatography.…”

“…We recently isolated and characterized five ficin (iso)forms (Azarkan et al, 2011). In the present work, we report the purification and characterization of an additional cysteine peptidase, termed ficin E. Purification of this novel protease was possible after depletion, by thiol-derivatization, of ficin A which co-eluted with ficin E on all the chromatographic media tested.…”

“…The properties of latex in plant defense and in alleviating various diseases have been attributed to the presence of several hydrolytic enzymes from which proteases play a key role. Most of the proteases found in latex of different plants belong to cysteine and serine proteases families (Singh et al, 2008;El Moussaoui et al, 2001;Azarkan et al, 2004Azarkan et al, , 2011Torres et al, 2012;Ramos et al, 2013;Yariswamy et al, 2013). Why latex contains high amounts of such proteases still remain enigmatic.…”

Optimization of Protease Extraction from Horse Mango (Mangifera foetidaLour) Kernels by a Response Surface Methodology

“…These include ficin from Ficus glabrata and Ficus laurifolia, 33,34) protease from Cynara cardunculus, 35,36) and the sodom apple (Calotropis procera), 37,38) and papain from papaya (Carica papaya). 39,40) Recent studies have indicated that sap from various varieties of Mangifera indica also exhibits both serine protease and cysteine protease activities.…”