Recent advances in molecular biology have extended our understanding of the metabolic processes related to microbial transformation of petroleum hydrocarbons. The physiological responses of microorganisms to the presence of hydrocarbons, including cell surface alterations and adaptive mechanisms for uptake and efflux of these substrates, have been characterized. New molecular techniques have enhanced our ability to investigate the dynamics of microbial communities in petroleum-impacted ecosystems. By establishing conditions which maximize rates and extents of microbial growth, hydrocarbon access, and transformation, highly accelerated and bioreactor-based petroleum waste degradation processes have been implemented. Biofilters capable of removing and biodegrading volatile petroleum contaminants in air streams with short substrate-microbe contact times ( 2 S and sulfoxides from petrochemical waste streams. Microbes also have potential for use in removal of nitrogen from crude oil leading to reduced nitric oxide emissions provided that technical problems similar to those experienced in biodesulfurization can be solved. Enzymes are being exploited to produce added-value products from petroleum substrates, and bacterial biosensors are being used to analyze petroleum-contaminated environments.

Recent Advances in Petroleum Microbiology

http://cau.ac.kr/~jjang14/BioMEMS/Lazcka_BSBE_Pathogen_Detection_Review_2006.pdf

Pathogen detection: a perspective of traditional methods and biosensors.

Soils represent a major sink for organic xenobiotic contaminants in the environment. The degree to which organic chemicals are retained within the soil is controlled by soil properties, such as organic matter, and the physico-chemical properties of the contaminant. Chemicals which display hydrophobic and lipophilic characteristics, as well as a recalcitrant chemical structure, will be retained within the soil, and depending on the `strength' of the association may persist for long periods of time. This review describes the behaviour of hydrophobic organic contaminants in soils, focusing on the mechanisms controlling interactions between soil and contaminants. The bioavailability of contaminants in soil is also discussed, particularly in relation to contact time with the soil. It considers the degradation of organic contaminants in soil and the mechanisms microbes use to access contaminants. Finally, the review discusses the `pros' and `cons' of chemical and biological techniques available for assessing bioavailability of hydrophobic organic chemicals in soils, highlighting the need to quantify bioavailability by chemical techniques. It concludes by highlighting the need for understanding the interactions between the soil, contaminants and biota which is crucial to understanding the bioavailability of contaminants in soils.

Bioavailability of hydrophobic organic contaminants in soils: fundamental concepts and techniques for analysis

Bacteria have long been the targets for genetic manipulation, but more recently they have been synthetically designed to carry out specific tasks. Among the simplest of these tasks is chemical compound and toxicity detection coupled to the production of a quantifiable reporter signal. In this Review, we describe the current design of bacterial bioreporters and their use in a range of assays to measure the presence of harmful chemicals in water, air, soil, food or biological specimens. New trends for integrating synthetic biology and microengineering into the design of bacterial bioreporter platforms are also highlighted.

Where microbiology meets microengineering: design and applications of reporter bacteria

Genetically engineered whole-cell sensing systems: coupling biological recognition with reporter genes.

A bacterial biosensor for benzene, toluene, and similar compounds has been constructed, characterized, and field tested on contaminated water and soil. The biosensor is based on a plasmid incorporating the transcriptional activator xylR from the TOL plasmid of Pseudomonas putida mt-2. The XylR protein binds a subset of toluene-like compounds and activates transcription at its promoter, Pu. A reporter plasmid was constructed by placing the luc gene for firefly luciferase under the control of XylR and Pu. When Escherichia coli cells were transformed with this plasmid vector, luminescence from the cells was induced in the presence of benzene, toluene, xylenes, and similar molecules. Accurate concentration dependencies of luminescence were obtained and exhibited K1/2 values ranging from 39.0 +/- 3.8 microM for 3-xylene to 2,690 +/- 160 microM for 3-methylbenzylalcohol (means +/- standard deviations). The luminescence response was specific for only toluene-like molecules that bind to and activate XylR. The biosensor cells were field tested on deep aquifer water, for which contaminant levels were known, and were able to accurately detect toluene derivative contamination in this water. The biosensor cells were also shown to detect BETX (benzene, toluene, and xylene) contamination in soil samples. These results demonstrate the capability of such a bacterial biosensor to accurately measure environmental contaminants and suggest a potential for its inexpensive application in field-ready assays.

/pdf/development-and-testing-of-a-bacterial-biosensor-for-toluene-1qtwsdzutr.pdf

Timothy A. Rand

Papers

Development and testing of a bacterial biosensor for toluene-based environmental contaminants