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Fig. S1. Thin-section (A) and negative-stain (B) electron microscopy of strain JH146. Each scale bar represents 200 nm.

Fig. S2. Phylogenetic sequence analysis of 16S rRNA (A), NifH (B), McrA (C), and ITS (D) from Methanocaldococcus strain JH146 and related species.

Fig. S3. Linear regression of number of cells per bottle plotted against the amount of CH4 per bottle for the growth of strain JH146 at 85°C and pH 7.0 [y = (4.644 × 106 cells μmol−1 CH4 produced)x − 1.86 × 107 cells tube−1, r2 = 0.969]. Growth yield (Y) was calculated from the slope of the regression line for each growth condition.

Table S1. Sequence similarity matrix for Methanocaldococcus strain JH146 compared with other known Methanocaldococcus gene sequences. NifH and McrA sequences were each compared at the amino acid level.

Table S2. Growth rate, cell yield based on methane production, cell-specific CH4 production rate, and cell-specific growth energy expenditure for Methanocaldococcus strain JH146 grown under varying conditions on otherwise standard growth medium. The error represents the 90% confidence interval (α = 0.1).

Table S3. Growth rate, cell yield based on methane production, cell-specific CH4 production rate, and cell-specific growth energy expenditure for Methanocaldococcus strain JH146 grown with varying concentrations of NH4Cl in otherwise nitrogen-free medium. The error represents the 90% confidence interval (α = 0.1).

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