목차

BACKGROUND...........................................................................................1
Properties of hydrocarbons, major components of petroleum oils and endosulfan.......................................................................................................1
Microbial degradation of petroleum oils and endosulfan.......................................................................................................4
REFERENCES..............................................................................................12
Chapter 1. Characterization of Petroleum Oil Degrading Bacteria from a Marine Bacterial Library Isolated from the Coastal Areas of Taean, Korea
Abstract.........................................................................................................22
1.1 INTRODUCTION....................................................................................24
1.1.1 The fate of petroleum oils in the environment…………..........24
1.1.2 Biodegradation of petroleum oils..………………........................25
1.1.3 Oil spill accident in Taean, Korea.............................................30
1.2 MATERIALS AND METHODS…………………………………...............32
1.2.1 Petroleum oil-contaminated sample collection from Taean, Korea......................................................................................32
1.2.2 Enrichment culture and Isolation of marine bacteria...............33
1.2.3 DNA extraction............................................................................33
1.2.4 PCR amplification and determination of 16S rRNA and alkane hydroxylase (alkB) gene sequences.........................................34
1.2.5 Phylogenetic analysis..................................................................36
1.2.6 Biochemical and physiological characterization of bacteria.....37
1.2.7 Analysis of cell growth pattern with crude oil..........................38
1.2.8 GC/MS analysis for the biodegradation of petroleum oils......39
1.2.9 Chemotaxonomic characterization of bacteria..........................39
1.3 RESULTS……………………..................................................................45
1.3.1 Marine bacterial library isolated from oil-contaminated sites of Taean..........................................................................................45
1.3.2 Phylogenetic analysis of the isolated bacteria.........................52
1.3.3 Petroleum oil degradation potential of the selected bacterial strains..........................................................................................59
1.3.4 Petroleum oil degradation potential of the constructed bacterial consortium...................................................................................61
1.3.5 Biochemical and physiological characterization of bacteria.....73
1.3.6 Fatty acid composition of the selected bacteria......................80
1.3.7 Identification and characterization of alkane hydroxylase (alkB) genes in the selected bacteria responsible for degradation of petroleum oils.............................................................................82
1.4 CONCLUSION………………..................................................................92
1.5 REFERENCES.......................................................................................93
Chapter 2. Characterization of Endosulfan Degrading Bacteria from a Marine Bacterial Library Isolated from Suncheon Bay, Korea
Abstract.......................................................................................................109
2.1 INTRODUCTION..................................................................................112
2.1.1 Organic chlorine pesticide, endosulfan...……………………....112
2.1.2 Effect of endosulfan on human health………….....................116
2.1.3 Biodegradation of endosulfan...................................................121
2.2 MATERIALS AND METHODS………………………………….............132
2.2.1 Sample collection from Suncheon Bay, Korea…………….....132
2.2.2 Chemicals………………………………………….........................133 2.2.3 Isolation of marine bacteria from the coastal wetland of Suncheon Bay............................................................................133 2.2.4 Enrichment culture and isolation of organic chlorine pesticide endosulfan-degrading marine bacteria....................................134
2.2.5 Construction of a marine bacterial consortium capable of endosulfan degradation............................................................135 2.2.6 Phylogenetic analysis of endosulfan-degrading bacteria........136
2.2.7 Biochemical and physiological characterization of endosulfan-degrading bacteria.............................................137
2.2.8 HPLC analysis of endosulfan-degrading bacterial strain EDB2 for the biodegradation of endosulfan.......................................138
2.2.9 Measurement of the removal of organic compounds by a constructed endosulfan-degrading bacterial consortium under several environmental conditions............................................139
2.2.10 Optimization of growth medium for the mass production of a microbial agent.......................................................................139 2.2.11 Batch and fed-batch culture of the consortium bacteria.....140
2.2.12 Viable count of the consortium bacterial culture for a freeze-dried microbial agent..................................................141
2.2.13 Formulation process of the bacterial consortium for the industrial production................................................................143 2.2.14 Evaluation of the formulated microbial agent for the removal of organic compounds under different temperature conditions................................................................................143
2.3 RESULTS ......................................……………………........................145
2.3.1 Marine bacterial library isolated from diverse sources of Suncheon Bay..........................................................................145
2.3.2 Marine bacteria from the synthetic water-contaminated sites...........................................................................................159
2.3.3 Marine bacteria from the endosulfan enrichment culture......161
2.3.4 Phylogenetic analysis of the bacteria from the synthetic water-contaminated sites.........................................................162
2.3.5 Phylogenetic analysis of the bacteria from the endosulfan enrichment culture....................................................................168
2.3.6 Biochemical and physiological characterization of the selected bacteria from endosulfan enrichment culture.........................172
2.3.7 Endosulfan degradation potential of the selected strain EDB2 (collection no. EM403).............................................................180
2.3.8 Construction of a marine bacterial consortium capable of endosulfan degradation............................................................182
2.3.9 Removal of organic compounds by the constructed consortium.................................................................................183
2.3.10 Effect of environmental conditions on the removal of organic compounds..............................................................................184
2.3.11 The optimized growth medium for the consortium bacteria and development of a liquid microbial agent ....................186
2.3.12 Removal of organic compounds under different temperature conditions by a liquid microbial agent..................................188
2.3.13 Development of a formulated microbial agent and evaluation for the removal of organic compounds................................189
2.4 CONCLUSION………………................................................................196
2.5 REFERENCES.....................................................................................198
ACKNOWLEDGMENTS..............................................................................216