Background/Objectives: Molybdenum, phenol and diesel are toxic to organism, and are part of global pollution. Their removal using microorganisms with multiple detoxification ability is being intensely sought as a cleaner and economic approach. Methods/Statistical analysis: A soil suspension was spread plated on a minimal salts media supplemented with molybdenum. Blue colonies, indicating molybdenum reduction was then screened for phenol and diesel degradation capabilities. Findings: A molybdenum-reducing bacterium locally isolated showed the ability to grow on phenol and diesel. The bacterium required pHs of between 5.8 and 6.3 and temperatures of between 30 and 40oC for optimal reduction. Among the carbon sources tested for supporting reduction, glucose was the best. A critical concentration of phosphate at just 5 mM was required, while molybdenum (sodium molybdate) was required between 15 and 25 mM. The absorption spectrum of the Mo-blue produced showed a characteristic maximum peak at 865 nm. The reduction of molybdenum was inhibited by the ions mercury, copper, chromium, lead and silver by 78.9, 78.4, 77.4, 53.5 and 36.8%, respectively. Analysis using phylogenetic analysis identifies the bacterium as Enterobacteriaceae bacterium strain DRY7. Growth on phenol and diesel as carbon sources showed that the optimal concentrations supporting growth was between 300 and 400 mg/L and between 300 and 500 mg/L, respectively. Application/Improvements: The capacity of this bacterium to detoxify a number of toxicants is an important property or bioremediation of soils contaminated with multiple toxicants.

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 MTT 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 IC 50 obtained by strain DRYS8 grown at the 12 hr-period in nutrient broth at 287deg;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. � Triveni Enterprises, Lucknow (India).

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® GN microplate panels and Microlog® 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° 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- μmax 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°C. The Mo-reducing enzyme was extremely sensitive to temperatures above 50°C (between 54 and 70°C). A plot of initial rates against substrate concentrations at 15 mM 12-MP registered a V m a x for NADH at 12.0 nmole Mo blue/min/mg protein. The apparent K m for NADH was 0.79 mM. At 5 mM NADH, the apparent V m a x and apparent K m values for 12-MP of 12.05 nmole/min/mg protein and 3.87 mM, respectively, were obtained. The catalytic efficiency (k c a t / K m) of the Mo-reducing enzyme was 5.47 M - 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.