Departamento Bioquímica y Biología Molecular y Edafología y Química Agrícola, Facultad de Ciencias, Universidad de Alicante, 03690 Alicante, Spain.
Department of Plant Biology, Faculty of Biology, Campus de Espinardo, University of Murcia, 30100 Murcia, Spain.
Biomolecules. 2023 Oct 16;13(10):1529. doi: 10.3390/biom13101529.
Here we present a study of the characterization and optimization of the production of trans-Resveratrol (-R) in grape ( cv. Gamay) cell cultures elicited with methyl jasmonate (MeJA) and dimethyl-β-cyclodextrin (DIMEB). The aim of this study was to determine the influence of a number of factors of the grapevine cell culture on -R production level in 250 mL shaken flasks that would enable the better control of this bioproduction system when it is upscaled to a 2 L stirred bioreactor. The factors included the optimal growth phase for elicitation, the concentration of elicitors and of biomass, the order of addition of elicitors, and the illumination regime and ageing of cells. We found out that the optimal biomass density for the production of -R was 19% (/) with an optimal ratio of 0.5 g DIMEB/g biomass. The most productive concentrations of the elicitors tested were 50 mM DIMEB and 100 µM MeJA, reaching maximum values of 4.18 mg·mL and 16.3 mg·g biomass of -R concentration and specific production, respectively. We found that the order of elicitor addition matters since, as compared with the simultaneous addition of both elicitors, the addition of MeJA 48 h before DIMEB results in ca. 40% less -R production, whilst there is no significant difference when MeJA is added 48 h after DIMEB. Upon upscaling, the better conditions tested for -R production were aeration at 1.7 vol/vol/min without agitation, 24 °C, and 30 g·L sucrose, achieving production rates similar to those obtained in shaken flasks.
我们在此介绍了一项利用甲基茉莉酸(MeJA)和二甲基-β-环糊精(DIMEB)诱导葡萄(品系 Gamay)细胞培养物来生产反式白藜芦醇(-R)的特征描述和优化研究。本研究的目的是确定许多葡萄细胞培养因素对 250 mL 摇瓶中 -R 产量的影响,以便在将该生物生产系统放大到 2 L 搅拌式生物反应器时更好地控制该系统。这些因素包括诱导的最佳生长阶段、诱导剂和生物量的浓度、诱导剂的添加顺序以及光照制度和细胞老化。我们发现,-R 生产的最佳生物量密度为 19%(/),最佳 DIMEB 与生物量的比例为 0.5 g DIMEB/g 生物质。测试的诱导剂最具生产力的浓度为 50 mM DIMEB 和 100 µM MeJA,分别达到 4.18 mg·mL 和 16.3 mg·g 生物质 -R 浓度和比生产的最大值。我们发现,诱导剂添加顺序很重要,因为与同时添加两种诱导剂相比,在 DIMEB 之前 48 小时添加 MeJA 会导致 -R 产量减少约 40%,而在 DIMEB 之后 48 小时添加 MeJA 则没有明显差异。在放大过程中,测试的最佳条件是在不搅拌的情况下以 1.7 体积/体积/分钟的速度通气,24°C 和 30 g·L 蔗糖,达到与摇瓶中获得的相似的生产速率。