Department of Pharmaceutical Technology, Laboratory of Microbial Technology, Federal Fluminense University, Niteroi, Rio de Janeiro, 24241 000, Brazil.
Department of Biochemistry, Laboratory of Oxidative Stress in Microorganisms, Rural Federal University of Rio de Janeiro, Seropédica, Rio de Janeiro, 23897-000, Brazil.
J Appl Microbiol. 2023 Aug 1;134(8). doi: 10.1093/jambio/lxad148.
Using in vitro assay and eukaryotic cell model of Saccharomyces cerevisiae, we investigated the impact of microbial fermentation on the antioxidant activity of phenolic substances.
Caffeic acid phenethyl ester (CAPE) and mangiferin were fermented by lactic acid bacteria (LAB), and the antioxidant activity of the fermented products was compared to that of the pure substances. This comparison was assessed using high-performance liquid chromatography (HPLC), in vitro by 2,2-Diphenyl-1-picrylhydrazyl (DPPH), and in vivo in yeast cells. The wild-type strain (BY4741) and its isogenic mutants in glutathione (Δgsh1), catalase (Δctt1), and superoxide dismutase (Δsod1) were treated with CAPE and mangiferin, fermented or not, and exposed to hydrogen peroxide (H2O2)-induced stress. The antioxidant activity was evaluated by cellular viability, intracellular oxidation, and lipid peroxidation. We expected that fermentation would change the antioxidant activity of phenolic substances. While HPLC analysis revealed changes in the composition of fermented products, significant alterations in antioxidant activity were only observed when using mutant strains. The fermentation of mangiferin increased dependency on GSH compared to the respective pure phenolic substance to resolve H2O2-induced stress. Additionally, CAPE appeared to act as a preconditioning agent, enhancing antioxidant responses, and promoting increased tolerance to H2O2 stress, and this mechanism was maintained after fermentation.
This study highlights that fermentation impacts the enzymatic mechanism of oxidative stress resolution, even though differences could not be observed in in vitro assays or in the wild-type strain.
利用体外测定和酿酒酵母真核细胞模型,研究微生物发酵对酚类物质抗氧化活性的影响。
利用乳酸菌对咖啡酸苯乙酯(CAPE)和芒果苷进行发酵,并比较发酵产物与纯物质的抗氧化活性。采用高效液相色谱法(HPLC)、2,2-二苯基-1-苦基肼(DPPH)体外法和酵母细胞内进行比较。采用野生型(BY4741)及其谷胱甘肽(Δgsh1)、过氧化氢酶(Δctt1)和超氧化物歧化酶(Δsod1)同源缺失突变株处理 CAPE 和芒果苷,进行或不进行发酵,并暴露于过氧化氢(H2O2)诱导的应激下。通过细胞活力、细胞内氧化和脂质过氧化来评估抗氧化活性。我们预计发酵会改变酚类物质的抗氧化活性。虽然 HPLC 分析显示发酵产物的组成发生了变化,但只有在用突变株进行分析时才观察到抗氧化活性的显著变化。与相应的纯酚类物质相比,芒果苷的发酵增加了对 GSH 的依赖性,以解决 H2O2 诱导的应激。此外,CAPE 似乎作为一种预处理剂,增强抗氧化反应,并促进对 H2O2 应激的耐受性增加,并且这种机制在发酵后仍能维持。
本研究强调了发酵会影响氧化应激解决的酶促机制,尽管在体外测定或野生型菌株中观察不到差异。