Hydrocarbon Degradation Potential of Halotolerant Bacteria

Index

1. Introduction
1.1 Objectives
1.2 Bioremediation
1.3 Hydrocarbon overview
1.4 Need for biodegradation
1.5 Approaches to biodegradation of hydrocarbons
1.6 Laboratory methods for studying hydrocarbon degradation
1.7 Microorganisms known to degrade hydrocarbons
1.8 Enzymes involved in hydrocarbon degradation
1.9 Factors affecting hydrocarbon degradation
1.10 Salinity and hydrocarbon degradation
1.11 Test organisms

2 Methodology
2.1 Materials
2.2 Instruments
2.3 Test organisms
2.4 Preliminary qualitative analysis
2.5 Cell lysis
2.6 Enzyme assay: catechol 2,3 dioxygenase
2.7 Enzyme assay: chlorocatechol 1,2-dioxygenase
2.8 Enzyme assay: protocatechuate 3,4-dioxygenase
2.9 Biofilm formation
2.10 Estimation of biofilm formation by crystal violet assay
2.11 Effect of microwave radiation on enzyme activity

3. Findings
3.1 Qualitative analysis
3.2 Enzyme activity
3.3 Effect of salinity on enzyme activity
3.4 Biofilm formation

4. Appendices

5. References

A. List of tables

Table 1.1 Major oil spills

Table 1.2 Halophiles known to degrade hydrocarbons

Table 1.3 Enzymes involved in biodegradation of petroleum

Table 2.1 Enzyme assay for catechol 2,3 dioxygenase

Table 2.2 Enzyme assay for chlorocatechol 1,2 dioxygenase

Table 2.3 Enzyme assay for protocatechuate 3,4-dioxygenase

Table 2.4 Classification of bacterial adhesion and biofilm formation

Table 3.1 Results of qualitative analysis

Table 3.2 Growth on combination of HC

Table 3.3 Activity of catechol 2,3 dioxygenase...

Table 3.4 Activity of chlorocatechol 1,2 dioxygenase

Table 3.5 Percent change in enzyme activity of V. salarius wrt P. oleovorans and their mixture

Table 3.6 Percent change in enzyme activity at different salt conc

Table 3.7 Results of biofilm formation

Table A1 Results of biofilm formation

B. List of figures

Fig 1.1 Catalytic cycle for intradiol cleavage

Fig 1.2 Catalytic cycle for extradiol cleavage

Fig 1.3 Ortho pathway for chlorocatechol degradation

Fig 1.4 General pathway for aromatic hydrocarbon degradation

Fig 3.1 Comparison of enzyme activity of catechol 2,3 dioxygenase at 6% salt concentration

Fig 3.2 Comparison of enzyme activity of chlorocatechol 1,2 dioxegenase at 6% salt concentration

Fig 3.3 Comparison of enzyme activity of catechol 2,3 dioxygenase at 10% salt concentration

Fig 3.4 Comparison of enzyme activity of chlorocatechol 1,2 dioxegenase at 10% salt concentration

Fig 3.5 Comparison of enzyme activity of V. salarius on different salt concentration

Fig3.6 Comparison of enzyme activity of P. oleovorans on different salt concentration

Fig 3.7 Comparison of enzyme activity mixture of V. salarius and P. oleovorans on different salt concentration

Fig 3.8 Comparison of enzyme activity of V. salarius on different salt concentration

Fig 3.9 Comparison of enzyme activity of P. oleovorans on different salt concentration

Fig 3.10 Comparison of enzyme activity mixture of V. salarius and P. oleovorans on different salt concentration34