Xu Jianlin, Trimble John J, Steinberg Lisa, Logan Bruce E
Department of Civil and Environmental Engineering, The Pennsylvania State University, 212 Sackett Building, University Park, PA 16802, USA.
Water Res. 2004 Feb;38(3):673-80. doi: 10.1016/j.watres.2003.10.017.
The effect of nitrate on perchlorate and chlorate reduction by perchlorate-respiring bacteria (PRB), and on chlorate reduction by chlorate-respiring bacteria (CRB), is not well understood, particularly with respect to the induction of pathways used to degrade these different chemicals. Based on kinetic data obtained in a series of batch tests, we determined that perchlorate respiratory enzymes were inducible (by chlorate or perchlorate) and separate from those used for denitrification by PRB strain Dechlorosoma sp. KJ. Aerobically grown cultures of KJ had lag times of greater than 0.3-2 days when transferred to a medium containing only perchlorate, chlorate, or nitrate as an electron acceptor. There were no lag times for transfers between identical media. Washed cells reduced very little nitrate (<10%) when grown only on chlorate or perchlorate. When grown on nitrate, they degraded little chlorate or perchlorate. The same lack of activity with these electron acceptors was also observed using cell extracts and methyl viologen as an electron carrier, indicating a lack of reactivity was not due to failure of the chemical to diffuse into the cell. Taken together, these results indicated that enzymes for perchlorate and nitrate reduction are separately expressed in strain KJ. The presence of small amounts of nitrate in contaminated groundwater may actually help to increase rates of perchlorate reduction once the nitrate is completely removed. When strain KJ was pre-grown on nitrate and perchlorate, perchlorate degradation (in the absence of nitrate) was more rapid compared to cells grown only on perchlorate. Pseudomonas sp. PDA was unable to degrade perchlorate or grow using nitrate, and the induction of enzymes necessary for chlorate respiration differed for strains KJ and PDA. While chlorate reductase and chlorite dismutase activity were induced in KJ by chlorate or perchlorate under anaerobic conditions, these two enzymes were constitutively expressed by PDA under anaerobic and aerobic conditions independent of the presence of chlorate. To our knowledge, this is the first report of constitutive expression of both chlorate reductase and chlorite dismutase in a bacterium.
硝酸盐对以高氯酸盐为呼吸底物的细菌(PRB)还原高氯酸盐和氯酸盐的影响,以及对以氯酸盐为呼吸底物的细菌(CRB)还原氯酸盐的影响,目前还不太清楚,特别是在用于降解这些不同化学物质的途径的诱导方面。基于一系列批次试验获得的动力学数据,我们确定高氯酸盐呼吸酶是可诱导的(由氯酸盐或高氯酸盐诱导),并且与PRB菌株Dechlorosoma sp. KJ用于反硝化的酶不同。KJ的需氧培养物在转移到仅含有高氯酸盐、氯酸盐或硝酸盐作为电子受体的培养基中时,滞后时间大于0.3 - 2天。在相同培养基之间转移没有滞后时间。仅在氯酸盐或高氯酸盐上生长时,洗涤后的细胞还原的硝酸盐非常少(<10%)。当在硝酸盐上生长时,它们降解的氯酸盐或高氯酸盐很少。使用细胞提取物和甲基紫精作为电子载体时,也观察到这些电子受体存在同样的活性缺乏,这表明活性缺乏不是由于化学物质无法扩散到细胞中。综上所述,这些结果表明高氯酸盐和硝酸盐还原酶在KJ菌株中是分别表达的。一旦硝酸盐被完全去除,受污染地下水中少量硝酸盐的存在实际上可能有助于提高高氯酸盐的还原速率。当KJ菌株在硝酸盐和高氯酸盐上预培养时,与仅在高氯酸盐上生长的细胞相比,(在没有硝酸盐的情况下)高氯酸盐的降解更快。假单胞菌属PDA无法降解高氯酸盐或利用硝酸盐生长,并且KJ菌株和PDA菌株用于氯酸盐呼吸所需酶的诱导情况不同。虽然在厌氧条件下,氯酸盐或高氯酸盐可诱导KJ菌株中的氯酸盐还原酶和亚氯酸盐歧化酶活性,但这两种酶在PDA菌株中在厌氧和好氧条件下均组成性表达,与氯酸盐的存在无关。据我们所知,这是首次报道一种细菌中氯酸盐还原酶和亚氯酸盐歧化酶均组成性表达。