Casimiro Bruno, Mota Inês, Veríssimo Paula, Canhoto Jorge, Correia Sandra
Centre for Functional Ecology, TERRA Associate Laboratory, Department of Life Sciences, Calçada Martim de Freitas, University of Coimbra, 3000-456 Coimbra, Portugal.
Center for Neuroscience and Cell Biology, University of Coimbra, Calçada Martim de Freitas, 3000-456 Coimbra, Portugal.
Plants (Basel). 2023 Jan 2;12(1):190. doi: 10.3390/plants12010190.
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% () 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.
植物细胞悬浮培养作为一种分析细胞和分子过程、代谢物合成及分化的工具被广泛应用，从而避开了植物结构的复杂性。在用于提高植物细胞代谢物产量的一系列方法中，最常用的方法之一是应用能够刺激与防御机制相关代谢途径的激发子。先前对番茄树细胞系和细胞悬浮培养物的蛋白质组学分析已用于在体外控制条件下进一步表征和优化生长及与胁迫相关的代谢物生产。这项工作的主要目标是开发一种新型的基于植物的生物反应器系统，采用激发方法生产水解酶。基于番茄树微繁殖和从诱导愈伤组织系建立植物细胞悬浮培养物的有效方案，优化了细胞生长，并表征了体外控制条件下的酶活性谱。通过测试不同的蔗糖浓度以及两种生物激发子的作用，发现液体培养基中3%（）的蔗糖浓度可提高水解酶的产量。此外，0.5和1.5 g/L的酪蛋白水解物促进了蛋白质的产生，而0.5 g/L的酵母提取物提高了糖苷酶活性。同时，0.05和0.1 g/L的壳聚糖提高了糖苷酶、碱性磷酸酶和蛋白酶的活性。
