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PROJECT TOPIC – EFFECT OF AN EXOGENOUS LIPASE ON BACTERIAL UTILIZATION OF PETROLEUM HYDROCARBON

EFFECT OF AN EXOGENOUS LIPASE ON BACTERIAL UTILIZATION OF PETROLEUM HYDROCARBON

 

ABSTRACT

Contaminated land is widely recognised as a potential threat to human health. The release of petroleum hydrocarbon through improper handling, storage and waste management is a major issue in environmental remediation. Despite regulatory steps that have been implemented to reduce, remove or eliminate production and release of these chemicals into the environment, significant environmental contamination has occurred in the past and will probably continue to occur in the future. Several studies have shown that microbial degradation of crude oil is an important factor contributing to the elimination of oil spills from the environment. Based on frequency of isolation, hydrocarbon degrading bacteria and fungi genera in soils and aquatic environment are
mainly the Bacillus, Micrococcus, Achromobacter, Nocardia, Pseudomonas, Trichoderma, Aspergillus, Mortierella and Penicillum species In the bioremediation of organically polluted sites, microbial extracellular enzyme activities are important and key steps, since only compounds with molecular mass lower than 600 daltons can pass through cell pores. The three common extracellular enzymes are lipase, amylase and protease.

In this study, petroleum hydrocarbon degrading bacteria were isolated from soil collected from the Mechanic Village in Nsukka. They were screened for lipase producing potential and the isolate with the best potential was selected for the study. Through morphological and biochemical tests, it was identified as Pseudomonas aeruginosa. Lipase was produced through submerge fermentation in a stationary phase. Microbial utilisation of petroleum hydrocarbon was monitored using the microorganism and the lipase. Results showed that the lipase was most active in the pH range 6.0 – 8.0 with an optimal reaction temperature of 40oC. Statistical analysis revealed that there was significant difference (P<0.05) between the controls and the test sample.

 

CHAPTER ONE

INTRODUCTION

1.1 OVERVIEW

Contamination of soil and groundwater through the release of petroleum hydrocarbons has been recognized as one of the principal issues in environmental remediation (Shah and Bhatt, 2006). Improper handling, storage and waste management practices have resulted in contamination of soil and aquifers; constituting threat to drinking water supplies, adversely affecting the people and many other areas of the environment (Fetter, 1993; Shah and Bhatt, 2006).
In many countries, the problems associated with contaminated sites are assuming an increasing prominence, and as such have become a worldwide problem (Vidali, 2001). In the oil producing areas of Nigeria, crude oil exploration has resulted in serious problems of oil contamination of the environment such as reduction in farming and fishing activities (Oluyege and Oluyemi, 2005). In the United States, it is estimated that most underground water tables are leaking (Kovalick, 1991). Despite regulatory steps that have been implemented to reduce, remove or eliminate production and release of these chemicals into the environment, significant environmental contamination has occurred in the past and will possibly continue to occur in the future (Baker and Diana, 1994).

The release of these pollutants in some cases such as industrial emissions are deliberate and well regulated; while in other cases such as chemical spills, they are accidental and largely unavoidable (Akpofure et al., 2000). Also, the activities of individuals at various mechanic workshops further contaminate the environment with petroleum products such as diesel, engine oil and petrol (De and Bello, 2002). Contaminated land is widely recognised as a potential threat to human health (Vidali, 2001). Although the components of petroleum derived products can be biodegraded easily in contrast to man-made compounds,  they are also dangerous (Wyszkowska et al, 2006). They display potential carcinogenic and mutagenic activities (Krahl et al., 2002). This recognition has resulted in international efforts to remedy contaminated sites, either as a response to the risk of adverse health or environmental effects caused by contamination or to enable such sites to be developed for use (Vidali, 2001).

Several studies have shown that microbial degradation of crude oil  is an important factor contributing to the elimination of oil spills from the environment (Okpokwasili and Amanchukwu, 1988; Fuentes et al., 1998; De and Bello, 2002). Based on frequency of isolation, hydrocarbon degrading bacteria and fungi genera in soils and aquatic environment are mainly the Bacillus, Micrococcus, Achromobacter, Nocardia, Pseudomonas, Trichoderma, Aspergillus,  ortierella and Penicillum species (Atlas, 1981;De and Bello, 2002). In the bioremediation of organically polluted sites, microbial extracellular enzyme activities are  important (Munster and De Haan, 1998), and key steps since only compounds with molecular mass lower than 600 daltons can pass through cell pores (Hoppe, 1991). The three groups of common extracellular enzymes are lipase, amylase and protease (Shah and Bhatt, 2006).

 

EFFECT OF AN EXOGENOUS LIPASE ON BACTERIAL UTILIZATION OF PETROLEUM HYDROCARBON

 

1.2 LIPASE

Lipases are glycerol ester hydrolases that catalyse the hydrolysis of ester linkages of glycerides at water-oil interface (Kashmiri et al., 2006). Even though this enzyme occurs widely in nature, only the microbial lipases are commercially significant (Sharma et al., 2001). They are amongst the most important biocatalysts carrying out novel reactions in both aqueous and non-aqueous media. This is as a result of their ability to utilise a wide range of substrates, high stability towards extremes of temperature, pH and organic solvents (Saxena et al., 1999). They catalyse the hydrolysis of triacylglycerols to glycerol and free fatty acids, and are activated only when adsorbed to an oil-water interface (Martinelle et al., 1995). However, the amount of oil available at the interface determines the activity of the lipase. This interface area can be increased substantially to its saturation limit by the use of emulsifiers as well as by agitation (Saxena et al., 1999). Lipases are important in a large number of industrial applications such as organic chemical processing, detergent formulations,  synthesis of biosurfactants, the oleochemical industries, the dairy industries and the agrochemical industries (Table 1).

 

EFFECT OF AN EXOGENOUS LIPASE ON BACTERIAL UTILIZATION OF PETROLEUM HYDROCARBON

 

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