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Protease-producing bacteria isolated from sub-Antarctic marine sediments of Isla de Los Estados (Argentina) were characterized, and the thermal inactivation kinetics of their extracellular proteases compared. Isolates were affiliated with the genera Pseudoalteromonas, Shewanella, Colwellia, Planococcus, and a strain to the family Flavobacteriaceae. Colwellia strains were moderate psychrophiles (optimal growth at about 15 degrees C, maximum growth temperature at around 25 degrees C). 16S rRNA phylogenetic analysis revealed that these strains and Colwellia aestuarii form a distinct lineage within the genus. The remaining isolates were psychrotolerant and grew optimally between 20 and 25 degrees C; two of them represent potentially novel species or genus (16S rRNA < 97% sequence similarity). The thermostability of the extracellular proteases produced by the isolates was analysed, and the inactivation rate constant (k (in)), the activation energy (Ea(in)) and the activation Gibbs free energy of thermal inactivation (Delta G( * ) (in)) determined. Delta G( * ) (in), calculated at 30 degrees C, varied between 97 and 124 kJ/mol. Colwellia enzyme extracts presented the highest thermosensitivity, while the most thermostable protease activity was shown by Shewanella spp. These results demonstrated that the stability to temperature of these enzymes varies considerably among the isolates, suggesting important variations in the thermal properties of the proteases that can coexist in this environment.

A Gram-positive, rod-shaped, spore-forming bacterium (PAT 05 T ) was isolated from the rhizosphere of the perennial shrub Atriplex lampa in north-eastern Patagonia, Argentina. Its overall biochemical and physiological characteristics indicated that this strain should be placed in the alkaliphilic Bacillus group. Strain PAT 05 T grew at pH 7-10 (optimum pH 8), but not at pH 6. Its DNA G+C content was 39?7 mol%. Sequence analysis of the 16S rRNA gene of PAT 05 T revealed the closest match (99?6 % similarity) with Bacillus sp. DSM 8714. The highest level of DNA-DNA relatedness (88?6 %) was also found with this strain. On the basis of 16S rRNA gene sequence similarity and phylogenetic analysis, G+C content and DNA-DNA hybridization data, strain PAT 05 T is related at the species level to Bacillus sp. DSM 8714, a member of a group referred as phenon 4a by Nielsen et al. [Nielsen, P., Fritze, D. & Priest, F. G. (1995). Microbiology 141, 1745-1761], which still lacks taxonomic standing. These results support the proposal of strain PAT 05 T (=DSM 16117 T =ATCC BAA-965 T ) as the type strain of Bacillus patagoniensis sp. nov.

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