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在氧和/或氯酸盐作为末端电子受体存在的情况下,在 Ideonella dechloratans 中表达亚氯酸盐歧化酶和氯酸盐还原酶。

Expression of chlorite dismutase and chlorate reductase in the presence of oxygen and/or chlorate as the terminal electron acceptor in Ideonella dechloratans.

机构信息

Department of Chemistry and Biomedical Sciences, Karlstad University, Karlstad, Sweden.

出版信息

Appl Environ Microbiol. 2012 Jun;78(12):4380-5. doi: 10.1128/AEM.07303-11. Epub 2012 Apr 6.

Abstract

The ability of microorganisms to perform dissimilatory (per)chlorate reduction is, for most species, known to be oxygen sensitive. Consequently, bioremediation processes for the removal of oxochlorates will be disturbed if oxygen is present. We measured the expression of chlorite dismutase and chlorate reductase in the presence of different terminal electron acceptors in the chlorate reducer Ideonella dechloratans. Enzyme activity assays and mRNA analyses by real-time quantitative reverse transcription (qRT)-PCR were performed on cell extracts from cells grown aerobically with and without chlorate and on cells grown anaerobically in the presence of chlorate. Our results showed that both chlorite dismutase and chlorate reductase are expressed during aerobic growth. However, transfer to anaerobic conditions with chlorate resulted in significantly enhanced enzyme activities and mRNA levels for both enzymes. Absence of oxygen was necessary for the induction to occur, since chlorate addition under aerobic conditions produced neither increased enzyme activities nor higher relative levels of mRNA. For chlorite dismutase, the observed increase in activity was on the same order of magnitude as the increase in the relative mRNA level, indicating gene regulation at the transcriptional level. However, chlorate reductase showed about 200 times higher enzyme activity in anaerobically induced cells, whereas the increase in mRNA was only about 10-fold, suggesting additional mechanisms influence the enzyme activity.

摘要

微生物进行异化(per)氯酸盐还原的能力,对于大多数物种来说,已知是对氧气敏感的。因此,如果存在氧气,去除氧氯酸盐的生物修复过程将会受到干扰。我们在氯酸盐还原菌 Ideonella dechloratans 中,测量了不同末端电子受体存在时亚氯酸歧化酶和氯酸盐还原酶的表达。通过实时定量逆转录(qRT)-PCR 对细胞提取物进行酶活性测定和 mRNA 分析,在有氧条件下生长的细胞和有氧条件下生长的细胞中进行氯酸盐处理,并在厌氧条件下生长的细胞中进行氯酸盐处理。我们的结果表明,在有氧生长过程中都表达了亚氯酸歧化酶和氯酸盐还原酶。然而,在有氯酸盐的厌氧条件下转移会导致两种酶的酶活性和 mRNA 水平显著增强。氧的缺失是诱导发生的必要条件,因为在有氧条件下添加氯酸盐既不会增加酶活性,也不会增加相对 mRNA 水平。对于亚氯酸歧化酶,观察到的活性增加与相对 mRNA 水平的增加处于同一数量级,表明基因在转录水平上受到调控。然而,在厌氧诱导的细胞中,氯酸盐还原酶的酶活性高出约 200 倍,而 mRNA 的增加仅约 10 倍,这表明其他机制会影响酶活性。

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