Dedysh Svetlana N, Knief Claudia, Dunfield Peter F
S. N. Winogradsky Institute of Microbiology, Russian Academy of Sciences, Moscow 117312, Russia.
J Bacteriol. 2005 Jul;187(13):4665-70. doi: 10.1128/JB.187.13.4665-4670.2005.
All aerobic methanotrophic bacteria described to date are unable to grow on substrates containing carbon-carbon bonds. Here we demonstrate that members of the recently discovered genus Methylocella are an exception to this. These bacteria are able to use as their sole energy source the one-carbon compounds methane and methanol, as well as the multicarbon compounds acetate, pyruvate, succinate, malate, and ethanol. To conclusively verify facultative growth, acetate and methane were used as model substrates in growth experiments with the type strain Methylocella silvestris BL2. Quantitative real-time PCR targeting the mmoX gene, which encodes a subunit of soluble methane monooxygenase, showed that copies of this gene increased in parallel with cell counts during growth on either acetate or methane as the sole substrate. This verified that cells possessing the genetic basis of methane oxidation grew on acetate as well as methane. Cloning of 16S rRNA genes and fluorescence in situ hybridization with strain-specific and genus-specific oligonucleotide probes detected no contaminants in cultures. The growth rate and carbon conversion efficiency were higher on acetate than on methane, and when both substrates were provided in excess, acetate was preferably used and methane oxidation was shut down. Our data demonstrate that not all methanotrophic bacteria are limited to growing on one-carbon compounds. This could have major implications for understanding the factors controlling methane fluxes in the environment.
迄今为止所描述的所有好氧甲烷氧化细菌都无法在含有碳 - 碳键的底物上生长。在此我们证明,最近发现的甲基孢囊菌属(Methylocella)成员是个例外。这些细菌能够将一碳化合物甲烷和甲醇以及多碳化合物乙酸盐、丙酮酸、琥珀酸盐、苹果酸盐和乙醇用作其唯一的能量来源。为了最终验证兼性生长,在以模式菌株森林甲基孢囊菌(Methylocella silvestris)BL2进行的生长实验中,将乙酸盐和甲烷用作模型底物。针对编码可溶性甲烷单加氧酶一个亚基的mmoX基因进行的定量实时PCR表明,在以乙酸盐或甲烷作为唯一底物生长期间,该基因的拷贝数与细胞计数平行增加。这证实了具有甲烷氧化遗传基础的细胞能够在乙酸盐以及甲烷上生长。16S rRNA基因的克隆以及与菌株特异性和属特异性寡核苷酸探针的荧光原位杂交检测到培养物中没有污染物。乙酸盐上的生长速率和碳转化效率高于甲烷,并且当两种底物都过量提供时,优先使用乙酸盐,甲烷氧化则停止。我们的数据表明,并非所有甲烷氧化细菌都局限于在一碳化合物上生长。这对于理解控制环境中甲烷通量的因素可能具有重大意义。