Farhan Ul Haque Muhammad, Kalidass Bhagyalakshmi, Bandow Nathan, Turpin Erick A, DiSpirito Alan A, Semrau Jeremy D
Department of Civil and Environmental Engineering, University of Michigan, Ann Arbor, Michigan, USA.
Roy J. Carver Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, Iowa, USA.
Appl Environ Microbiol. 2015 Nov;81(21):7546-52. doi: 10.1128/AEM.02542-15. Epub 2015 Aug 21.
Methanotrophs have multiple methane monooxygenases that are well known to be regulated by copper, i.e., a "copper switch." At low copper/biomass ratios the soluble methane monooxygenase (sMMO) is expressed while expression and activity of the particulate methane monooxygenase (pMMO) increases with increasing availability of copper. In many methanotrophs there are also multiple methanol dehydrogenases (MeDHs), one based on Mxa and another based on Xox. Mxa-MeDH is known to have calcium in its active site, while Xox-MeDHs have been shown to have rare earth elements in their active site. We show here that the expression levels of Mxa-MeDH and Xox-MeDH in Methylosinus trichosporium OB3b significantly decreased and increased, respectively, when grown in the presence of cerium but the absence of copper compared to the absence of both metals. Expression of sMMO and pMMO was not affected. In the presence of copper, the effect of cerium on gene expression was less significant, i.e., expression of Mxa-MeDH in the presence of copper and cerium was slightly lower than in the presence of copper alone, but Xox-MeDH was again found to increase significantly. As expected, the addition of copper caused sMMO and pMMO expression levels to significantly decrease and increase, respectively, but the simultaneous addition of cerium had no discernible effect on MMO expression. As a result, it appears Mxa-MeDH can be uncoupled from methane oxidation by sMMO in M. trichosporium OB3b but not from pMMO.
甲烷氧化菌具有多种甲烷单加氧酶,众所周知这些酶受铜的调控,即存在一个“铜开关”。在低铜/生物量比时,可溶性甲烷单加氧酶(sMMO)表达,而颗粒性甲烷单加氧酶(pMMO)的表达和活性则随着铜可利用性的增加而增加。在许多甲烷氧化菌中还存在多种甲醇脱氢酶(MeDHs),一种基于Mxa,另一种基于Xox。已知Mxa-MeDH的活性位点含有钙,而Xox-MeDHs的活性位点已被证明含有稀土元素。我们在此表明,与两种金属都不存在的情况相比,在存在铈但不存在铜的条件下培养时,甲基弯曲菌OB3b中Mxa-MeDH和Xox-MeDH的表达水平分别显著降低和升高。sMMO和pMMO的表达不受影响。在存在铜的情况下,铈对基因表达的影响较小,即存在铜和铈时Mxa-MeDH的表达略低于仅存在铜时,但Xox-MeDH再次被发现显著增加。正如预期的那样,添加铜分别导致sMMO和pMMO的表达水平显著降低和升高,但同时添加铈对MMO的表达没有明显影响。因此,在甲基弯曲菌OB3b中,Mxa-MeDH似乎可以与sMMO介导的甲烷氧化解偶联,但不能与pMMO介导的甲烷氧化解偶联。