Ruminant digestive tract microbes hydrolyse plant biomass, and the application of metagenomic techniques can provide good coverage of their glycosyl hydrolase enzymes. A metagenomic library of circa 70,000 fosmids was constructed from bacterial DNA isolated from bovine rumen and subsequently screened for cellulose hydrolysing activities on a CMC agar medium. Two clones were selected based on large clearance zones on the CMC agar plates. Following nucleotide sequencing, translational analysis and homology searches, two cellulase encoding genes (cel5A and cel5B) belonging to the glycosyl hydrolyse family 5 were identified. Both genes encoded pre-proteins of about 62 kDa, containing signal leader peptides which could be cleaved to form mature proteins of about 60 kDa. Biochemical characterisation revealed that both enzymes showed alkaline pH optima of 9.0 and the temperature optima of 65 °C. Substrate specificity profiling of the two enzymes using 1,4-β-D-cello- and xylo-oligosaccharides revealed preference for longer oligosaccharides (n ≥ 3) for both enzymes, suggesting that they are endo-cellulases/xylanases. The bifunctional properties of the two identified enzymes render them potentially useful in degrading the β-1,4 bonds of both the cellulose and hemicellulose polymers.
Pit latrines are the most commonly used sanitation systems in many developing countries. Various researchers have reported elevated nitrate concentrations in groundwater in the vicinity of pit latrines and this could pose a serious health risk to the users of the water source. Faecal sludge from pit latrines contains high concentrations of nitrogen and organic matter (3-5 g•ℓ −1 N and 20-50 g•ℓ −1 COD); however, it is produced at a very low rate (1.5 ℓ•capita −1 •d −1) relative to that of waterborne sewage systems. A pit latrine basically only confines the waste and no real treatment takes place. In this research the nitrogen was removed in a biological filter using a combination of nitrification and denitrification processes. The aim of this investigation was to determine the effect of air supplied at different rates, namely, 0, 0.3, 1.0 and 2.0 m 3 •h −1 N, on the biological filtration process. The application rate was 0.04 m 3 •m −2 •d −1. More than 90% removal of nitrogen was observed at an air supply rate of 1.0 m 3 •h −1 N. At lower air supply rates nitrification was not complete. At an air supply rate of 2.0 m 3 •h −1 nitrogen removal was also approx. 90%, but the biological filter only became stable after about 2 months of operation, possibly due to desiccation of the biomass.
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