Most of petrochemical oil pollutants contain aliphatic lipid and aromatic hydrocarbon compounds that are hydrophobic to water but sticky to surface of certain materials. These widespread fuel oil pollutants (total petroleum hydrocarbon,TPH) are difficult to be removed and need to be degraded with specific microorganisms associated with abundant quantity of extracellular polymer and secreted biosurfactant. Biosurfactant with the extracellular polysaccharide and the polypeptide promoted biofilm can be developed from diverse microbial consortia that are most found in hydrocarbon contaminated soil. With the secreted biosurfactants dissolving and penetrating into the oil layer, a biodegradation process can be enhanced with the enriched microorganisms which can drill channels for mass transport of nutrients and bioproducts. In order to carry out a systematic environmental molceular biotechnological (SEMBT) remediation, three research groups are established: 1) bioagent development with molecular biotechnology, (2) molecular biomonitoring technology, and 3) biotechnology integration for environmental engineering. Group 1 with 3 professors’ laboratories will identify specific bacterial strains (indigenic consortia) from the local environment, then grow up large quantity of these bioagents through scaled-up fermentation processes. Group 2 processors’ laboratories will develop different biomonitoring technologies, which include microarray to identify the specific strains, and DNA extraction to identify the diverse microbial community in a contaminated soil. Group 3 with 2 professors’ laboratories will develop biofilm technologies: to aggregate the diverse microorganisms as biofilm consortia, along with the intergration of all of the biotechnology established by these laboratories in this project. This research group will work out the systematic technologies of the bioagent augmentation, the biomonitoring, and the biofilm stimulation to perform an useful bioremediation process (SEMBT) in a petroleum contaminated site. Within one year, a set of ex-situ bioremediation demonstration will be established with a pilot scale plant of 20 m3 biopile treating the total petroleum hydrocarbon (TPH). These biotechnologies of ex-situ bioremediation will be transferred to the Taiwan China Petroleum Corporation, and the CHC environmental engineering firm for scale-up application in the near future. The project goal is to demonstrate a systematic environemental molecular biotechnology (SEMBT) for biodegradation technology that is feasible for remediating a large area of hydrocarbon comtaminated site.