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A fundamental and principal difficulty of the future energy supply is that the formation of fossil fuels is much slower than the rate of their exploitation. Therefore the reserves which can be recovered in an energetically feasible manner are shrinking parallel with an increasing world-wide energy demand. Among the alternative energy carriers, hydrogen is preferred because it is easy to transport and store and it burns to environmentally friendly water vapour when utilized. Hydrogen can be produced in biological systems, however, our understanding of the molecular details is just emerging.

Two methanotrophic bacteria with optimum growth temperatures above 40 degrees C were isolated. Thermotolerant strain LK6 was isolated from agricultural soil, and the moderately thermophilic strain OR2 was isolated from the effluent of an underground hot spring. When compared to the described thermophilic methanotrophs Methylococcus capsulatus and Methylococcus thermophilus, these strains are phenotypically similar to Methylococcus thermophilus. However, their 16S rRNA gene sequences are markedly different from the sequence of Methylococcus thermophilus ( approximately 8% divergence) and, together with Methylomonas gracilis, they form a distinct, new genus within the gamma-subgroup of the Proteobacteria related to extant Type I methanotrophs. Further phenotypic characterisation showed that the isolates possess particulate methane monooxygenase (pMMO) but do not contain soluble methane monooxygenase. The nucleotide sequence of a gene encoding pMMO (pmoA) was determined for both isolates and for Methylomonas gracilis. PmoA sequence comparisons confirmed the monophyletic nature of this newly recognised group of thermophilic methanotrophs and their relationship to previously described Type I methanotrophs. We propose that strains OR2 and LK6, together with the misclassified thermophilic strains Methylomonas gracilis VKM-14LT and Methylococcus thermophilus IMV-B3122, comprise a new genus of thermophilic methanotrophs, Methylocaldum gen. nov., containing three new species: Methylocaldum szegediense, Methylocaldum tepidum and Methylocaldum gracile.

Methanotrophic bacteria were isolated from a natural environment potentially favorable to heat-tolerant methanotrophs. An improved colony plate assay was developed and used to identify putative methanotrophic colonies with high confidence. Fourteen new isolates were purified and partially characterized. These new isolates exhibit a DNA sequence homology of up to 97% with the conserved regions in the mmoX and mmoC genes of the soluble methane monooxygenase (MMO)-coding gene cluster of Methylococcus capsulatus Bath. The copper regulation of soluble MMO expression in the same isolates, however, differs from that of M. capsulatus Bath, as the new isolates can tolerate up to 0.8 microM copper without loss of MMO activity while a drastic reduction of MMO activity occurs already at 0.1 microM copper in M. capsulatus Bath. The isolates can be cultivated and utilized at elevated temperatures, and their copper- and heat-tolerant MMO activity makes these bacteria ideal candidates for future biotechnological use.

Methanotroph microorganisms oxidize methane in four steps, producing methanol, formaldehyde, formate intermediers and eventually degrade methane to carbon dioxide and water. It is possible to separate the pathway into four steps in the cell free extract or after partial purification of the various enzymes. The key enzyme is a metalloenzyme, methane monooxygenase (MMO) which catalyses the oxidation of methane to methanol. MMO is also capable of biodegrading exceptionally harmful and stable chlorinated hydrocarbons. Produced by various industrial activities, most chlorinated hydrocarbons are toxic, potential and/or proven carcinogens and their decomposition challenges water treatment technologies.