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在连续培养的欧洲亚硝化单胞菌细胞中添加硫酸镉后merA、trxA、amoA和hao的表达情况。

Expression of merA, trxA, amoA, and hao in continuously cultured Nitrosomonas europaea cells exposed to cadmium sulfate additions.

作者信息

Radniecki Tyler S, Semprini Lewis, Dolan Mark E

机构信息

School of Chemical, Biological and Environmental Engineering, Oregon State University, Corvallis, Oregon 97331-3212, USA.

出版信息

Biotechnol Bioeng. 2009 Dec 1;104(5):1004-11. doi: 10.1002/bit.22454.

Abstract

The effects of CdSO(4) additions on the gene expressions of a mercury reductase, merA, an oxidative stress protein, trxA, the ammonia-monooxygenase enzyme (AMO), amoA, and the hydroxylamine oxidoreductase enzyme (HAO), hao, were examined in continuously cultured N. europaea cells. The reactor was fed 50 mM NH(4)+ and was operated for 78 days with a 6.9 days hydraulic retention time. Over this period, six successive batch additions of CdSO(4) were made with increasing maximum concentrations ranging from 1 to 60 microM Cd(2+). The expression of merA was highly correlated with the level of Cd(2+) within the reactor (Rs = 0.90) with significant up-regulation measured at non-inhibitory Cd(2+) concentrations. Cd(2+) appears to target AMO specifically at lower concentrations and caused oxidative stress at higher concentrations, as indicated by the SOURs (specific oxygen uptake rates) and the up-regulation of trxA. Since Cd(2+) inhibition is irreversible and amoA was up-regulated in response to Cd(2+) inhibition, it is hypothesized that de novo synthesis of the AMO enzyme occurred and was responsible for the observed recovery in activity. Continuously cultured N. europaea cells were more resistant to Cd(2+) inhibition than previously examined batch cultured cells due to the presence of Mg(2+) and Ca(2+) in the growth media, suggesting that Cd(2+) enters the cell through Mg(2+) and Ca(2+) import channels. The up-regulation of merA during exposure to non-inhibitory Cd(2+) levels indicates that merA is an excellent early warning signal for Cd(2+) inhibition.

摘要

在连续培养的欧洲亚硝化单胞菌(N. europaea)细胞中,研究了添加硫酸镉(CdSO₄)对汞还原酶基因merA、氧化应激蛋白基因trxA、氨单加氧酶(AMO)基因amoA以及羟胺氧化还原酶(HAO)基因hao表达的影响。反应器中加入50 mM NH₄⁺,在水力停留时间为6.9天的条件下运行78天。在此期间,连续六次分批添加CdSO₄,最大浓度从1 μM Cd²⁺增加到60 μM Cd²⁺。merA的表达与反应器内Cd²⁺水平高度相关(Rs = 0.90),在非抑制性Cd²⁺浓度下可检测到显著上调。如特定氧摄取率(SOURs)和trxA的上调所示,Cd²⁺在较低浓度下似乎特异性靶向AMO,而在较高浓度下会引起氧化应激。由于Cd²⁺抑制是不可逆的,且amoA在Cd²⁺抑制作用下上调,因此推测发生了AMO酶的从头合成,并导致了观察到的活性恢复。由于生长培养基中存在Mg²⁺和Ca²⁺,连续培养的欧洲亚硝化单胞菌细胞比之前研究的分批培养细胞对Cd²⁺抑制更具抗性,这表明Cd²⁺通过Mg²⁺和Ca²⁺的导入通道进入细胞。在暴露于非抑制性Cd²⁺水平期间merA的上调表明merA是Cd²⁺抑制的良好早期预警信号。

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