Department of Biological Science , Sungkyunkwan University , Suwon , South Korea.
Department of Marine Resource Development , Gangneung-Wonju National University , Gangneung , South Korea.
Environ Sci Technol. 2019 Jul 2;53(13):7840-7850. doi: 10.1021/acs.est.9b01460. Epub 2019 Jun 19.
Glutathione S-transferases (GSTs) play an important role in phase II of detoxification to protect cells in response to oxidative stress generated by exogenous toxicants. Despite their important role in defense, studies on invertebrate GSTs have mainly focused on identification and characterization. Here, we isolated omega and sigma classes of GSTs from the freshwater rotifer Brachionus calyciflorus and the marine rotifer Brachionus koreanus and explored their antioxidant function in response to metal-induced oxidative stress. The recombinant Bc- and Bk-GSTs were successfully transformed and expressed in Escherichia coli. Their antioxidant potential was characterized by measuring kinetic properties and enzymatic activity in response to pH, temperature, and chemical inhibitor. In addition, a disk diffusion assay, reactive oxygen species assay, and morphological analysis revealed that GST transformed into E. coli significantly protected cells from oxidative stress induced by HO and metals (Hg, Cd, Cu, and Zn). Stronger antioxidant activity was exhibited by GST-S compared to GST-O in both rotifers, suggesting that GST-S plays a prominent function as an antioxidant defense mechanism in Brachionus spp. Overall, our study clearly shows the antioxidant role of Bk- and Bc-GSTs in E. coli and provides a greater understanding of GST class-specific and interspecific detoxification in rotifer Brachionus spp.
谷胱甘肽 S-转移酶(GSTs)在解毒的第二期反应中发挥重要作用,以保护细胞免受外源性毒物产生的氧化应激。尽管 GSTs 在防御中具有重要作用,但对无脊椎动物 GSTs 的研究主要集中在鉴定和特性描述上。在这里,我们从淡水轮虫褶皱臂尾轮虫和海洋轮虫秀丽白虾中分离出ω和σ类 GSTs,并探索了它们在应对金属诱导的氧化应激时的抗氧化功能。成功地将重组的 Bc-和 Bk-GSTs 转化并在大肠杆菌中表达。通过测量动力学特性和对 pH 值、温度和化学抑制剂的酶活性来表征它们的抗氧化潜力。此外,圆盘扩散试验、活性氧测定和形态分析表明,转化为大肠杆菌的 GST 显著保护细胞免受 HO 和金属(Hg、Cd、Cu 和 Zn)诱导的氧化应激。在两种轮虫中,GST-S 比 GST-O 表现出更强的抗氧化活性,表明 GST-S 在褶皱臂尾轮虫属中作为抗氧化防御机制发挥着突出的作用。总的来说,我们的研究清楚地表明了 Bk-和 Bc-GSTs 在大肠杆菌中的抗氧化作用,并提供了对轮虫褶皱臂尾轮虫属 GST 类特异性和种间解毒作用的更深入了解。