Respiratory activity inhibition by toxic compounds in bacteria and yeast has been used to detect toxic compounds in the environment. Often the age of culture contributes towards the sensitivity of detection. In the present work, the effect of growth period on the sensitivity of an inhibitive assay for heavy metals using bacterial respiratory assay system based on the reduction of the water soluble tetrazolium dye MU is reported. A silver-sensitive isolate was discovered to exhibit different sensitivities towards silver at different growth periods. An exponential decay model adequately described the inhibition due to silver. Analysis using ANOVA with post-hoc Tukey's test showed that the IC50 obtained by strain DRYS8 grown at the 12 hr-period in nutrient broth at 28 degrees C gave the lowest value compared to other growth periods. This study highlights the importance of taking into accounts growth conditions and age of culture in developing cellular-based bioassays.

A diesel-degrading bacterium was isolated from a diesel-contaminated site in Selangor, Malaysia. The isolate was tentatively identified as Acinetobacter sp. strain DRY12 based on partial 16S rDNA molecular phylogeny and Biolog (R) GN microplate panels and Microlog (R) database. Optimum growth occurred from 3 to 5% diesel and the strain was able to tolerate as high as 8% diesel. The optimal pH that supported growth of the bacterium was between pH 7.5 to 8.0. The isolate exhibited optimal growth in between 30 and 35 degrees C. The best nitrogen source was potassium nitrate (between 0.6 and 0.9% (w/v)) followed by ammonium chloride, sodium nitrite and ammonium sulphate in descending order. An almost complete removal of diesel components was seen from the reduction in hydrocarbon peaks observed using Solid Phase Microextraction Gas Chromatography analysis after 10 days of incubation. The best growth kinetic model to fit experimental data was the Haldane model of substrate inhibiting growth with a correlation coefficient value of 0.97. The maximum growth rate- p(max) was 0.039 hr(-1) while the saturation constant or half velocity constant Ks and inhibition constant Ki, were 0.387% and 4.46%, respectively. MATH assays showed that 75% of the bacterium was found in the hexadecane phase indicating that the bacterium was hydrophobic. The characteristics of this bacterium make it useful for bioremediation works in the Tropics.

The first purification of the Mo-reducing enzyme from Serratia sp. strain DRY5 that is responsible for molybdenum reduction to molybdenum blue in the bacterium is reported. The monomeric enzyme has an apparent molecular weight of 105 kDalton. The isoelectric point of this enzyme was 7.55. The enzyme has an optimum pH of 6.0 and maximum activity between 25 and 35 degrees C. The Mo-reducing enzyme was extremely sensitive to temperatures above 50 degrees C (between 54 and 70 degrees C). A plot of initial rates against substrate concentrations at 15mM 12-MP registered a V-max for NADH at 12.0 nmole Mo blue/min/mg protein. The apparent k(m) for NADH was 0.79mM. At 5mM NADH, the apparent V-max and apparent k(m) values for 12-MP of 12.05 nmole/min/mg protein and 3.87mM, respectively, were obtained. The catalytic efficiency (k(cat)/k(m)) of the Mo-reducing enzyme was 5.47M(-1) s(-1). The purification of this enzyme could probably help to solve the phenomenon of molybdenum reduction to molybdenum blue first reported in 1896 and would be useful for the understanding of the underlying mechanism in molybdenum bioremediation involving bioreduction.