International Journal of Systematic and Evolutionary Microbiology, Vol 52, 1799-1806, Copyright © 2002 by Society for General Microbiology

Methanocalculus taiwanensis sp. nov., isolated from an estuarine environment

M. C. Lai, S. C. Chen, C. M. Shu, M. S. Chiou, C. C. Wang, M. J. Chuang, T. Y. Hong, C. C. Liu, L. J. Lai and J. J. Hua
Department of Botany, National Chung Hsing University, Taichung, 40227 Taiwan, Republic of China

Two novel hydrogenotrophic methanogens, designated strains P2F9704a(T) and P2F9705, were isolated from an estuary in Eriln Shi, Taiwan. The cells of strain P2F9704a(T) were non-motile, irregular cocci 0.9--1.4 microm in diameter. They stained Gram-negative. The cells catabolized formate and H(2)+CO(2) to produce methane, but did not utilize acetate, methanol, trimethylamine, ethanol or secondary alcohols as methanogenic substrates. The optimal growth parameters for strain P2F9704a(T) were pH 6.7, 37 degrees C and 0.5% NaCl. Acetate was required for cell growth even though it was not a substrate for methanogenesis. The trace element tungsten was not required but slightly stimulated the growth of strain P2F9704a(T). However, tungsten extended the growth ranges relating to temperature, pH and salt. The sequences of the 16S rRNA genes of strains P2F9704a(T) and P2F9705 were nearly identical and possessed 99.1 and 98.5% similarity to the genes of Methanocalculus pumilus and Methanocalculus halotolerans, respectively. In addition, strain P2F9704a(T) possessed 14 and 12% DNA relatedness with respect to Methanocalculus pumilus and Methanocalculus halotolerans, respectively. In addition, the optimal salt concentrations, the cellular protein profiles and the molecular masses of surface-layer protein subunits of strain P2F9704a(T) were different from those of the other two known Methanocalculus species. On the basis of these observations, it is proposed that these two organisms should be placed in a new species, namely Methanocalculus taiwanensis. The type strain is P2F9704a(T) (=OCM 671(T)=CCRC 16182(T)=DSM 14663(T)).

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