Department of Biochemistry, Faculty of Medical Science, Naresuan University, Phitsanulok, 65000, Thailand.
Department of Biology, Faculty of Science, Naresuan University, Phitsanulok, 65000, Thailand.
J Biotechnol. 2021 Jan 20;326:28-36. doi: 10.1016/j.jbiotec.2020.12.006. Epub 2020 Dec 28.
Obtaining large-scale hairy root cultures is a major challenge to increasing root biomass and secondary metabolite production. Enhanced production of stilbene compounds such as trans-resveratrol, trans-arachidin-1 and trans-arachidin-3 was achieved using an elicitor treatment procedure. Two different hairy root inoculum densities were investigated and compared between shake flask and bioreactor cultures. The lowest growth index was observed using a 20 g/L inoculum size in the bioreactor, which differed significantly from bioreactor of 5 g/L. Increasing the hairy root inoculum size from 5 g/L to 20 g/L in both the shake flask and bioreactor significantly improve antioxidant activity, phenolic content and stilbene compound levels. The highest ABTS and FRAP antioxidant activity, and levels of total phenolic compounds, trans-arachidin-1 and trans-arachidin-3 in the crude extract were demonstrated in shake flask cultures with a 20 g/L inoculum after elicitation for 72 h. The minimum inhibitory concentrations (MICs) of the crude extract to inhibit growth of foodborne microbes, S. aureus, S. typhimurium and E. coli, were 187.5, 250 and 500 μg/mL, respectively. This was due to the ability of the crude extract to disrupt the cell membrane, as observed by scanning electron microscopy (SEM) showing ruptured pores on the S. aureus and S. typhimurium cell surfaces. Moreover, the E. coli cell division process could be inhibited by the crude extract, which promoted an increase in cell size. A DNA nicking assay indicated that a 50 μg/mL concentration of the crude extract caused plasmid DNA damage that might be due to a genotoxic effect of the pro-oxidant activity of the crude extract.
获得大规模的发根培养物是增加根生物量和次生代谢产物生产的主要挑战。通过使用诱导剂处理程序,实现了白藜芦醇、反式-芝麻素-1 和反式-芝麻素-3 等芪类化合物的产量增加。研究了两种不同的发根接种物密度,并在摇瓶和生物反应器培养物之间进行了比较。在生物反应器中,接种物大小为 20 g/L 时观察到最低的生长指数,与生物反应器中 5 g/L 时的生长指数显著不同。在摇瓶和生物反应器中,将发根接种物大小从 5 g/L 增加到 20 g/L,均显著提高了抗氧化活性、酚含量和芪类化合物水平。在经过 72 小时诱导后,摇瓶培养物中接种物为 20 g/L 时,粗提物的 ABTS 和 FRAP 抗氧化活性最高,总酚含量、反式-芝麻素-1 和反式-芝麻素-3 的水平最高。粗提物对食源性病原体金黄色葡萄球菌、鼠伤寒沙门氏菌和大肠杆菌生长的最小抑菌浓度(MIC)分别为 187.5、250 和 500 μg/mL。这是由于粗提物能够破坏细胞膜,扫描电子显微镜(SEM)观察到金黄色葡萄球菌和鼠伤寒沙门氏菌细胞表面出现破裂的孔。此外,粗提物可以抑制大肠杆菌的细胞分裂过程,从而促进细胞体积增大。DNA 切口试验表明,浓度为 50 μg/mL 的粗提物导致质粒 DNA 损伤,这可能是由于粗提物的促氧化活性产生的遗传毒性作用。