Water assessment using ultra-weak bioluminescence.

In this paper a method to evaluate the presence of microorganisms of the coliform group in water samples using the ultra-weak bioluminescence (UWB) is proposed. A series of UWB measurements and optical density measurements from cultures of both a set of standard E. coli strain samples, and a set of water samples from a river near Curitiba City in Brazil were performed. All samples were previously incubated at 37°C for 11h in nutritive medium before the temporal UWB emission profiles data were acquired for a period of 24h inside a dark chamber of an especially implemented instrumentation capable of doing photon counting measurements. For the optical density measurements, a spectrophotometer was used to acquire the growth kinetics of those cultures for a period of 13h, and the results compared to the UWB profiles. Periodic time-components analysis of the UWB data from both the set of standard E. coli samples and the set of the river's water samples were performed and compared to each other. The results have shown that the UWB temporal profiles resemble in some way the growth kinetics curve and the periodic time-components analysis is an effective way to discriminate between contaminated and non-contaminated samples, therefore the method may be viable for detecting coliforms in water samples in less time than usual methods.

Isolation of a novel lipase from a metagenomic library derived from mangrove sediment from the south Brazilian coast.

A novel gene coding for a LipA-like lipase with 283 amino acids and a molecular mass of 32 kDa was isolated and characterized from a metagenomic library prepared from mangrove sediment from the south Brazilian coast. LipA was 52% identical to a lipolytic enzyme from an uncultured bacterium and shared only low identities (< or =31%) with lipases/esterases from cultivable microorganisms. Phylogenetic analysis showed that LipA, together with an orthologous protein from an uncultured bacterium, forms a unique branch within family I of true lipases, thereby constituting a new lipase subfamily. Activity determination using crude extracts of Escherichia coli bearing the lipA gene revealed that this new enzyme has a preference for esters with short-chain fatty acids (C < or = 10) and has maximum activity against p-nitrophenyl-caprate (chain length C10, 0.87 U/mg protein). The optimum pH of LipA was 8.0, and the enzyme was active over a temperature range of 20 to 35 degrees C, with optimum activity against p-nitrophenyl-butyrate at 35 degrees C and pH 8.0.