Autor(es): Casimiro, Bruno ; Mota, Inês ; Veríssimo, Paula ; Canhoto, Jorge M. P. L. ; Correia, Sandra
Data: 2023
Identificador Persistente: https://hdl.handle.net/10316/114910
Origem: Estudo Geral - Universidade de Coimbra
Projeto/bolsa: info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB/PT;
Assunto(s): cell growth; chitosan; elicitation; glycosidase activity; hydrolytic enzymes
Plant cell suspension cultures are widely used as a tool for analyzing cellular and molecular processes, metabolite synthesis, and differentiation, bypassing the structural complexity of plants. Within the range of approaches used to increase the production of metabolites by plant cells, one of the most recurrent is applying elicitors capable of stimulating metabolic pathways related to defense mechanisms. Previous proteomics analysis of tamarillo cell lines and cell suspension cultures have been used to further characterize and optimize the growth and stress-related metabolite production under in vitro controlled conditions. The main objective of this work was to develop a novel plant-based bioreactor system to produce hydrolytic enzymes using an elicitation approach. Based on effective protocols for tamarillo micropropagation and plant cell suspension culture establishment from induced callus lines, cell growth has been optimized, and enzymatic activity profiles under in vitro controlled conditions characterized. By testing different sucrose concentrations and the effects of two types of biotic elicitors, it was found that 3% (w/v) sucrose concentration in the liquid medium enhanced the production of hydrolytic enzymes. Moreover, casein hydrolysate at 0.5 and 1.5 g/L promoted protein production, whereas yeast extract (0.5 g/L) enhanced glycosidase activity. Meanwhile, chitosan (0.05 and 0.1 g/L) enhanced glycosidases, alkaline phosphates, and protease activities.
