PROIMI-CONICET, Av. Belgrano y Pje. Caseros, Tucumán T4001MVB, Argentina; Facultad de Ciencias Naturales, Universidad Nacional de Salta, Salta, Argentina; INIQUI-CONICET, Av. Bolivia 5150, Salta 4400, Argentina.
PROIMI-CONICET, Av. Belgrano y Pje. Caseros, Tucumán T4001MVB, Argentina.
Ecotoxicol Environ Saf. 2018 Feb;148:490-500. doi: 10.1016/j.ecoenv.2017.10.076. Epub 2017 Nov 6.
Cyberlindnera jadinii M9 and Wickerhamomyces anomalus M10 isolated from textile-dye liquid effluents has shown capacity for chromium detoxification via Cr(VI) biological reduction. The aim of the study was to evaluate the effect of hexavalent chromium on synthesis of novel and/or specific proteins involved in chromium tolerance and reduction in response to chromium overload in two indigenous yeasts. A study was carried out following a proteomic approach with W. anomalus M10 and Cy. jadinii M9 strains. For this, proteins extracts belonging to total cell extracts, membranes and mitochondria were analyzed. When Cr(VI) was added to culture medium there was an over-synthesis of 39 proteins involved in different metabolic pathways. In both strains, chromium supplementation changed protein biosynthesis by upregulating proteins involved in stress response, methionine metabolism, energy production, protein degradation and novel oxide-reductase enzymes. Moreover, we observed that Cy. jadinii M9 and W. anomalus M10 displayed ability to activate superoxide dismutase, catalase and chromate reductase activity. Two enzymes from the total cell extracts, type II nitroreductase (Frm2) and flavoprotein wrbA (Ycp4), were identified as possibly responsible for inducing crude chromate-reductase activity in cytoplasm of W. anomalus M10 under chromium overload. In Cy.jadinii M9, mitochondrial Ferredoxine-NADP reductase (Yah1) and membrane FAD flavoprotein (Lpd1) were identified as probably involved in Cr(VI) reduction. To our knowledge, this is the first study proposing chromate reductase activity of these four enzymes in yeast and reporting a relationship between protein synthesis, enzymatic response and chromium biospeciation in Cy. jadinii and W. anomalus.
从纺织染料废液中分离出的 Cyberlindnera jadinii M9 和 Wickerhamomyces anomalus M10 具有通过 Cr(VI)生物还原解毒铬的能力。本研究的目的是评估六价铬对两种本土酵母中参与铬耐受和还原的新型和/或特异性蛋白质合成的影响。采用蛋白质组学方法对 W. anomalus M10 和 Cy. jadinii M9 菌株进行了研究。为此，分析了总细胞提取物、膜和线粒体的蛋白质提取物。当向培养基中添加 Cr(VI)时，会过度合成 39 种参与不同代谢途径的蛋白质。在两种菌株中，铬的补充通过上调参与应激反应、甲硫氨酸代谢、能量产生、蛋白质降解和新型氧化还原酶的蛋白质来改变蛋白质的生物合成。此外，我们观察到 Cy. jadinii M9 和 W. anomalus M10 表现出激活超氧化物歧化酶、过氧化氢酶和铬酸盐还原酶活性的能力。从总细胞提取物中鉴定出两种酶，即硝基还原酶 II (Frm2)和黄素蛋白 wrbA (Ycp4)，可能负责在 W. anomalus M10 细胞质中诱导铬酸盐还原酶活性在铬超载下。在 Cy.jadinii M9 中，鉴定出线粒体 Ferredoxine-NADP 还原酶 (Yah1)和膜 FAD 黄素蛋白 (Lpd1)可能参与 Cr(VI)还原。据我们所知，这是首次提出这四种酶在酵母中的铬酸盐还原酶活性，并报道 Cy. jadinii 和 W. anomalus 中蛋白质合成、酶反应和铬生物形态之间的关系。
