Selection of the Optimal Medium for Adsorption of Plant Proteases

Immobilized enzymes are the most sought-after preparations in the global market. They are used in medicine, veterinary medicine, the food industry, winemaking and brewing. The simplest method for immobilizing biocatalysts on insoluble carriers is the simple adsorption method. Its advantage is that it preserves the natural conformation of the enzyme, which slightly reduces its catalytic ability compared to the native form. In our study, we carried out the selection of optimal conditions for adsorption immobilization of acid-soluble chitosan (Mr = 350 kDa) enzymes of plant origin (ficin, papain and bromelain) on a matrix. Ficin (EC 3.4.22.3), papain (EC 3.4.22.2) and bromelain (EC 3.4.22.4) (Sigma) were chosen as the objects of study, azocasein (Sigma) was used as a substrate for hydrolysis and an acid-soluble high-molecular-weight chitosan (350 kDa) was used as an immobilization matrix, synthesized by Bioprogress CJSC. Suitable buffer systems for immobilization were identified by the optimal ratio of protein content and total and specific activity. Ficin is immobilized on a chitosan matrix using glycine buffer with a pH of 8.6. Glycine buffer with a pH of 8.6–10.5 is an optimal medium for sorption of papain on chitosan. Bromelain is immobilized on a chitosan matrix under Tris-glycine buffer with pH 8.5 conditions.

PSI relieves the pressure of membrane fusion

Some plant proteases contain a latent sequence known as the plant-specific insert (PSI) that, upon release from the full protease sequence, initiates membrane fusion to defend from pathogens. However, the mechanism by which it exerts its effects has been unclear. Zhao et al. report an elegant integration of biophysical experiments and molecular dynamics simulations to reveal events leading up to PSI-mediated membrane fusion. Their results demonstrate a pH-dependent monomer-to-dimer transition, clear evidence of membrane association, and probable structures of prefusion intermediates. These data expand our understanding of the elusive PSIs and may provide new directions for antimicrobial development.

Characterization of Ficus benjamina and Artocarpus heterophyllus Proteases as Potential Rennet Alternatives

Rennet, a milk coagulant exhibiting proteolytic activity, is a crucial component in cheese industries. Its price and availability have discouraged the growth of some small scale cheese industries. Therefore, an alternative for rennet will be beneficial for the industries. Among other sources, plant proteases offer some advantages as rennet alternatives. This study aimed to investigate the potential of plant proteases obtained from the latex as potential rennet alternatives. A total of six plants from the genus Ficus and Artocarpus were screened for their proteolytic activity and milk coagulating ability. The screening indicated that all six tested plants displayed proteolytic activity at various levels, but only Ficus benjamina and Artocarpus heterophyllus produced a firm milk curd. Hence, both F. benjamina and A. heterophyllus were determined to be the most potential. Further characterizations suggested that F. benjamina and A. heterophyllus protease were optimum at pH 7.0 also at 50°C and 40°C, respectively. At their optimum conditions, both proteases exhibited a lower MCA/PA ratio than that of the rennet. This study contributed to scientific knowledge development by becoming the first to characterize the optimum conditions of F. benjamina and A. heterophyllus’ proteases, investigate their MCA/PA ratio, and compare their activity against commercial rennet. The examination of their potentials as rennet alternatives could benefit small cheese industries and the communities.

The Role of Proteases in Determining Stomatal Development and Tuning Pore Aperture: A Review

Plant proteases, the proteolytic enzymes that catalyze protein breakdown and recycling, play an essential role in a variety of biological processes including stomatal development and distribution, as well as, systemic stress responses. In this review, we summarize what is known about the participation of proteases in both stomatal organogenesis and on the stomatal pore aperture tuning, with particular emphasis on their involvement in numerous signaling pathways triggered by abiotic and biotic stressors. There is a compelling body of evidence demonstrating that several proteases are directly or indirectly implicated in the process of stomatal development, affecting stomatal index, density, spacing, as well as, size. In addition, proteases are reported to be involved in a transient adjustment of stomatal aperture, thus orchestrating gas exchange. Consequently, the proteases-mediated regulation of stomatal movements considerably affects plants’ ability to cope not only with abiotic stressors, but also to perceive and respond to biotic stimuli. Even though the determining role of proteases on stomatal development and functioning is just beginning to unfold, our understanding of the underlying processes and cellular mechanisms still remains far from being completed.

Plant Aspartic Proteases for Industrial Applications: Thistle Get Better

Plant proteases have a number of applications in industrial processes including cheese manufacturing. The flower of the cardoon plant (Cynara cardunculus L.) is traditionally used as a milk-clotting agent in protected designation of origin cheeses made from goat and sheep milk. Plant-derived rennets are of particular importance to consumers who wish to eat cheeses that are produced without harming any animals. In this review, we have highlighted the importance of plant proteases, particularly aspartic proteases, in industrial processes, as well as exploring more fundamental aspects of their synthesis. We have also reviewed and discussed the production of these enzymes using sustainable and cost-effective alternative platforms.