Effect of insertional mutations in the pueA and pueB genes encoding two polyurethanases in Pseudomonas chlororaphis contained within a gene cluster.

AIMS: To better understand the role of PueA and PueB from Pseudomonas chlororaphis in polyurethane degradation, the present study was conducted to create insertional mutants in their respective genes. METHODS AND RESULTS: Growth kinetic studies showed that the pueA knockout mutant had a greater effect than the pueB knockout mutant. The pueA mutant had an 80% decrease in cell density from that of the wild type, while the pueB mutant had an 18% decrease in cell density. Polyurethane utilization followed Michaelis-Menten kinetics. The pueA and pueB mutants exhibited a 17% and 10% decrease respectively in growth rate using polyurethane when compared with the wild type. CONCLUSIONS: In this present study, pueA and pueB, are shown to be part of an ABC transporter gene cluster that consists of seven open reading frames. Mutational analysis results suggest that PueA may play a more major role in polyurethane degradation than PueB based on cell density and growth rates. SIGNIFICANCE AND IMPACT OF THE STUDY: The results from this study provide a starting point for the eventual enhancement and bioremediation of polyurethane waste. Understanding the role of polyurethane-degrading enzymes is useful for the creation of strains for this purpose.

Phytoremediation of small-scale oil spills in fresh marsh environments: a mesocosm simulation.

Research was conducted to assess the impact of oiling on fresh-marsh plant communities and to test the efficacy of techniques that may be used to enhance the bioremediation of crude oil spills in these environments while minimizing secondary anthropogenic impacts. To emulate field conditions, a mesocosm facility was used that houses 120 mesocosm vessels, each of 200-1 capacity. A five-way factorial treatment arrangement was used that included two substrates (inorganic, organic), two nutrient regimes (fertilized, not fertilized), two aeration levels (substrate aeration, no aeration), three oiling concentrations (0-, 5-, 10-1 m(-2) of South Louisiana Sweet Crude oil), and four vascular plant species (Alternanthera philoxeroides, Panicum hemitomon, Phragmites australis, Sagittaria lancifolia, and an unplanted control). Under the 5- and 10-1 m(-2) oiling concentrations, S. lancifolia displayed a short-term response of increased productivity, whereas P. hemitomon had the highest biomass production and photosynthetic rates at the end of the 18-month experiment. Overall plant growth and productivity, as well as oil degradation, were significantly higher in the inorganic substrate, indicating that biodegradation of oil spills in organic substrates may require a longer time period. Time-released fertilizer also stimulated plant productivity and resulted in higher soil respiratory quotients, suggestive of greater microbial activity, particularly in aerated mesocosms. The amount of oil remaining after 18 months was lowest in aerated and fertilized mesocosms containing either P. hemitomon or S. lancifolia and a substrate of low organic matter content.

Diauxic Growth of Azotobacter vinelandii on Galactose and Glucose: Regulation of Glucose Transport by Another Hexose.

The growth curve of Azotobacter vinelandii was biphasic when the organism was grown in a medium containing a mixture of galactose and glucose. Galactose was the primary carbon source; glucose was also consumed, but the rate at which it was consumed was lower than the rate at which galactose was consumed during the first phase of growth. Metabolic pathways for both sugars were induced. Cell cultures exhibited a second lag period as galactose was depleted. The length of this lag phase varied from 2 to 10 h depending on the pregrowth history of the cells. The second log growth phase occurred at the expense of the remaining glucose in the medium and was accompanied by induction of the high-maximum rate of metabolism glucose-induced glucose permease and increases in the levels of glucose metabolic enzymes. The second lag phase of diauxie may have been due to the time required for induction of the glucose-induced glucose permease.

Comparison of the presence-absence and membrane filter techniques for coliform detection in small, nonchlorinated water distribution systems.

The traditional membrane filter (American Public Health Association, Standard Methods for the Examination of Water and Wastewater, 16th ed., American Public Health Association, Washington, D.C., 1985) and presence-absence (P-A) (J. A. Clark, Can. J. Microbiol. 14:13-18, 1968) techniques for the detection of coliform bacteria were compared in a small nonchlorinated drinking water distribution system by using total positive samples and frequency-of-occurrence analyses. No significant differences (P less than 0.05) were found in detection of the presence of coliform bacteria or in changes in the frequency of occurrence with time. A reduction in P-A sample volume (to 50 ml) was not found to statistically affect the comparative results of traditional membrane filter and P-A tests.