Chan Kam Che. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 199-219). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstracts --- p.iii / 摘要 --- p.v / Contents --- p.vii / List of figures --- p.xiv / List of tables --- p.xix / Abbreviations --- p.xxii / Chapter Chapter I --- Introduction --- p.1 / Chapter 1.1 --- Persistent organic pollutants --- p.1 / Chapter 1.2 --- DDT and DDE --- p.2 / Chapter 1.2.1 --- Background --- p.2 / Chapter 1.2.2 --- Health effects --- p.4 / Chapter 1.2.3 --- Environmental exposure of DDE --- p.4 / Chapter 1.2.4 --- Level of DDE in human --- p.9 / Chapter 1.2.5 --- Biodegradation of DDE --- p.10 / Chapter 1.3 --- Remediation methods --- p.11 / Chapter 1.3.1 --- Physical/ chemical treatment --- p.11 / Chapter 1.3.2 --- Bioremediation --- p.13 / Chapter 1.4 --- Fungal Bioremediation --- p.14 / Chapter 1.5 --- Ligninolytic enzymes --- p.15 / Chapter 1.5.1 --- Laccase --- p.15 / Chapter 1.5.2 --- Peroxidases --- p.20 / Chapter 1.5.2.1 --- Manganese Peroxidase (MnP) --- p.20 / Chapter 1.5.2.1 --- Lignin Peroxidase (LiP) --- p.24 / Chapter 1.6 --- Cultivation of Pleurotus pulmonarius --- p.27 / Chapter 1.7 --- Enzyme technology on environmental cleanup and its limitation --- p.28 / Chapter 1.8 --- Aims and objectives of this study --- p.29 / Chapter Chapter II --- Materials and Methods --- p.30 / Chapter 2.1 --- Organism and growth conditions --- p.30 / Chapter 2.2 --- Cultivation and the expression of the ligninolytic enzyme-coding genes during solid-state-fermentation of edible mushroom Pleurotus pulmonarius --- p.30 / Chapter 2.3 --- Treatment of DDE by living P. pulmonarius --- p.31 / Chapter 2.3.1 --- Optimization of DDE removal in broth system --- p.31 / Chapter 2.3.1.1 --- Effects of initial DDE concentration on the removal of DDE --- p.32 / Chapter 2.3.1.2 --- Effects of inoculum size on the removal of DDE --- p.33 / Chapter 2.3.1.3 --- Effects of incubation time on the removal of DDE and transcriptional profiles of the ligninolytic enzyme-coding genes --- p.33 / Chapter 2.3.2 --- Optimization of DDE removal in soil system --- p.34 / Chapter 2.3.2.1 --- Effects of initial DDE concentration on the removal of DDE --- p.34 / Chapter 2.3.2.2 --- Effects of inoculum size on the removal of DDE --- p.35 / Chapter 2.3.2.3 --- Effects of incubation time on the removal of DDE --- p.35 / Chapter 2.3.2.4 --- Transcription of the ligninolytic enzyme-coding genes --- p.35 / Chapter 2.4 --- Treatment of DDE by 1st SMC of p. pulmonarius grown on straw-based compost --- p.36 / Chapter 2.4.1 --- Optimization of DDE removal in soil system --- p.36 / Chapter 2.5 --- Treatment of DDE by crude enzyme preparations of P. pulmonarius grown on straw-based compost --- p.36 / Chapter 2.5.1 --- Optimization of DDE removal in broth system --- p.36 / Chapter 2.5.1.1 --- Effects of initial DDE concentration on the removal of DDE --- p.37 / Chapter 2.5.1.2 --- Effects of amounts of crude enzyme preparations on the removal of DDE --- p.37 / Chapter 2.5.1.3 --- Effects of incubation time on the removal of DDE --- p.37 / Chapter 2.5.2 --- Optimization of DDE removal in soil system --- p.37 / Chapter 2.5.2.1 --- Effects of initial DDE concentration on the removal of DDE --- p.38 / Chapter 2.5.2.2 --- Effects of amount of crude enzyme preparations on the removal of DDE --- p.38 / Chapter 2.5.2.3 --- Effects of incubation time on the removal of DDE --- p.38 / Chapter 2.6 --- Soil characterization --- p.39 / Chapter 2.6.1 --- Identification of organic contaminants in soil sample from Gene Garden using Gas Chromatography/Mass Spectrometry (GC/MS) --- p.39 / Chapter 2.6.2 --- Determination of soil texture --- p.42 / Chapter 2.6.3 --- Fresh soil/air-dried sample moisture --- p.44 / Chapter 2.6.4 --- "Soil pH, electrical conductivity & salinity" --- p.44 / Chapter 2.6.5 --- Total organic carbon contents --- p.44 / Chapter 2.6.6 --- Total nitrogen and total phosphorus --- p.44 / Chapter 2.6.7 --- Available nitrogen --- p.45 / Chapter 2.6.8 --- Available phosphorus --- p.45 / Chapter 2.6.9 --- Potassium value --- p.46 / Chapter 2.7 --- Quantification of residual DDE level --- p.47 / Chapter 2.7.1 --- Preparation of DDE stock solution --- p.47 / Chapter 2.7.2 --- Extraction and quantification of DDE using Gas Chromatography with Electron Capture Detector (GC/μECD) --- p.47 / Chapter 2.7.3 --- Identification of DDE breakdown products by GC/MS --- p.50 / Chapter 2.8 --- Extraction of protein and ligninolytic enzymes --- p.53 / Chapter 2.8.1 --- Protein assay --- p.53 / Chapter 2.8.2 --- Laccase assay --- p.53 / Chapter 2.8.3 --- Manganese peroxidase assay --- p.54 / Chapter 2.8.4 --- Calculation of activity and specific activity of laccase and manganese peroxidase --- p.54 / Chapter 2.9 --- Estimation of fungal biomass --- p.55 / Chapter 2.9.1 --- Preparation of ergosterol standard solution --- p.56 / Chapter 2.9.2 --- Analysis of ergosterol content --- p.56 / Chapter 2.10 --- Expression of the ligninolytic enzyme-coding genes --- p.58 / Chapter 2.10.1 --- Preparation of ribonuclease free reagents and apparatus --- p.58 / Chapter 2.10.2 --- RNA isolation and purification --- p.58 / Chapter 2.10.3 --- cDNA synthesis --- p.59 / Chapter 2.10.4 --- Semi-quantification of ligninolytic enzyme-coding gene expression by RT-PCR --- p.59 / Chapter 2.11 --- Preparation of crude enzyme preparations from P. pulmonarius compost --- p.63 / Chapter 2.12 --- "Assessment criteria: removal efficiency, RE, and removal capacity, RC" --- p.63 / Chapter 2.13 --- Statistical analysis “ --- p.64 / Chapter Chapter III --- Results --- p.65 / Chapter 3.1 --- Soil characterization --- p.65 / Chapter 3.2 --- Cultivation and the expression of the ligninolytic enzyme-coding genes during solid-state-fermentation of edible mushroom Pleurotus pulmonarius --- p.66 / Chapter 3.2.1 --- Mushroom yield --- p.66 / Chapter 3.2.2 --- Protein content --- p.66 / Chapter 3.2.3 --- Specific ligninolytic enzymes activities --- p.66 / Chapter 3.2.4 --- Ergosterol content --- p.69 / Chapter 3.2.5 --- Ligninolytic enzymes productivities --- p.69 / Chapter 3.2.6 --- Expression of the ligninolytic enzyme-coding genes during solid-state-fermentation --- p.72 / Chapter 3.3 --- Treatment of DDE by living P. pulmonaruis --- p.78 / Chapter 3.3.1 --- Optimization of DDE removal in broth system --- p.78 / Chapter 3.3.1.1 --- Effects of initial DDE concentration on the removal of DDE --- p.78 / Chapter 3.3.1.1.1 --- Effects of DDE on biomass development --- p.78 / Chapter 3.3.1.1.2 --- Protein content --- p.78 / Chapter 3.3.1.1.3 --- Specific ligninolytic enzyme activities --- p.78 / Chapter 3.3.1.1.4 --- Ligninolytic enzyme productivities --- p.79 / Chapter 3.3.1.1.5 --- DDE removal and removal capacity --- p.79 / Chapter 3.3.1.2 --- Effects of inoculum sizes on the removal of DDE --- p.84 / Chapter 3.3.1.2.1 --- Effects of DDE on biomass development --- p.84 / Chapter 3.3.1.2.2 --- Protein content --- p.84 / Chapter 3.3.1.2.3 --- Specific ligninolytic enzyme activities --- p.85 / Chapter 3.3.1.2.4 --- Ligninolytic enzyme productivities --- p.85 / Chapter 3.3.1.2.5 --- DDE removal and removal capacity --- p.85 / Chapter 3.3.1.3 --- Effects of incubation time on the removal of 4.0 mM DDE/g biomass --- p.89 / Chapter 3.3.1.3.1 --- Effects of DDE on biomass development --- p.89 / Chapter 3.3.1.3.2 --- Protein content --- p.89 / Chapter 3.3.1.3.3 --- Specific ligninolytic enzyme activities and ligninolytic enzyme productivities --- p.89 / Chapter 3.3.1.3.4 --- DDE removal and removal capacity --- p.90 / Chapter 3.3.1.3.5 --- Putative degradation derivatives --- p.90 / Chapter 3.3.1.3.6 --- Expression of the ligninolytic enzyme-coding genes during the removal of 4.0 mM DDE/g biomass --- p.94 / Chapter 3.3.1.4 --- Effects of incubation time on the removal of 10.0 mM DDE/g biomass --- p.100 / Chapter 3.3.1.4.1 --- Effects of DDE on biomass development --- p.100 / Chapter 3.3.1.4.2 --- Protein content --- p.100 / Chapter 3.3.1.4.3 --- Specific ligninolytic enzyme activities and ligninolytic enzyme productivities --- p.100 / Chapter 3.3.1.4.4 --- Expression of the ligninolytic enzyme-coding genes during the removal of 10.0 mM DDE/g biomass --- p.102 / Chapter 3.3.2 --- Optimization of DDE removal in soil system --- p.107 / Chapter 3.3.2.1 --- Effects of initial DDE concentration on the removal of DDE --- p.107 / Chapter 3.3.2.1.1 --- Ergosterol content --- p.107 / Chapter 3.3.2.1.2 --- Protein content --- p.107 / Chapter 3.3.2.1.3 --- Specific ligninolytic enzyme activities and ligninolytic enzyme productivities --- p.107 / Chapter 3.3.2.1.4 --- DDE removal and removal capacity --- p.108 / Chapter 3.3.2.2 --- Effects of inoculum sizes on the removal of DDE --- p.111 / Chapter 3.3.2.2.1 --- Ergosterol content --- p.111 / Chapter 3.3.2.2.2 --- Protein content --- p.111 / Chapter 3.3.2.2.3 --- Specific ligninolytic enzyme activities and ligninolytic enzyme productivities --- p.111 / Chapter 3.3.2.2.4 --- DDE removal and removal capacity --- p.112 / Chapter 3.3.2.3 --- Effects of incubation time on the removal of DDE --- p.115 / Chapter 3.3.2.3.1 --- Ergosterol content --- p.115 / Chapter 3.3.2.3.2 --- Protein content --- p.115 / Chapter 3.3.2.3.3 --- Specific ligninolytic enzyme activities and ligninolytic enzyme productivities --- p.115 / Chapter 3.3.2.3.4 --- DDE removal and removal capacity --- p.116 / Chapter 3.3.2.3.5 --- Putative degradation derivatives --- p.116 / Chapter 3.3.2.4 --- Transcription of the ligninolytic enzyme-coding genes --- p.121 / Chapter 3.4 --- Treatment of DDE by 1st SMC of p. pulmonarius grown on straw-based compost --- p.127 / Chapter 3.4.1 --- Optimization of DDE removal in soil system --- p.127 / Chapter 3.4.1.1 --- Effects of initial DDE concentration on the removal of DDE --- p.127 / Chapter 3.4.1.1.1 --- Ergosterol content --- p.127 / Chapter 3.4.1.1.2 --- Protein content --- p.127 / Chapter 3.4.1.1.3 --- Specific ligninolytic enzyme activities and ligninolytic enzyme productivities --- p.127 / Chapter 3.4.1.1.4 --- DDE removal and removal capacity --- p.128 / Chapter 3.4.1.2 --- Effects of inoculum sizes on the removal of DDE --- p.132 / Chapter 3.4.1.2.1 --- Ergosterol content --- p.132 / Chapter 3.4.1.2.2 --- Protein content --- p.132 / Chapter 3.4.1.2.3 --- Specific ligninolytic enzyme activities and ligninolytic enzyme productivities --- p.132 / Chapter 3.4.1.2.4 --- DDE removal and removal capacity --- p.133 / Chapter 3.4.1.3 --- Effects of incubation time on the removal of DDE --- p.136 / Chapter 3.4.1.3.1 --- Ergosterol content --- p.136 / Chapter 3.4.1.3.2 --- Protein content --- p.136 / Chapter 3.4.1.3.3 --- Specific ligninolytic enzyme activities and ligninolytic enzyme productivities --- p.136 / Chapter 3.4.1.3.4 --- DDE removal and removal capacity --- p.137 / Chapter 3.4.1.3.5 --- Putative degradation derivatives --- p.137 / Chapter 3.5 --- Treatment of DDE by crude enzyme preparations of P. pulmonarius grown on straw-based compost --- p.142 / Chapter 3.5.1 --- The crude enzyme preparations of P. pulmonarius grown on straw-based compost --- p.142 / Chapter 3.5.2 --- Optimization of DDE removal in broth system --- p.143 / Chapter 3.5.2.1 --- Effects of initial DDE concentration on the removal of DDE --- p.143 / Chapter 3.5.2.2 --- Effects of amounts of crude enzyme preparations on the removal of DDE --- p.145 / Chapter 3.5.2.3 --- Effects of incubation time on the removal of DDE --- p.147 / Chapter 3.5.2.4 --- Putative degradation derivatives --- p.147 / Chapter 3.5.3 --- Optimization of DDE removal in soil system --- p.151 / Chapter 3.5.3.1 --- Effects of initial DDE concentration on the removal of DDE --- p.151 / Chapter 3.5.3.2 --- Effects of amounts of crude enzyme preparations on the removal of DDE --- p.151 / Chapter 3.5.3.3 --- Effects of incubation time on the removal of DDE --- p.154 / Chapter 3.5.3.4 --- Putative degradation derivatives --- p.154 / Chapter Chapter IV --- Discussions --- p.158 / Chapter 4.1 --- Quantification of the expression of the ligninolytic enzyme-coding genes --- p.158 / Chapter 4.2 --- Artificial cultivation and the expression of the ligninolytic enzyme-coding genes during solid-state-fermentation of edible mushroom Pleurotus pulmonarius --- p.164 / Chapter 4.3 --- Treatment of DDE by living P. pulmonarius --- p.166 / Chapter 4.3.1 --- Optimization of DDE removal in broth system --- p.166 / Chapter 4.3.2 --- Optimization of DDE removal in soil system --- p.169 / Chapter 4.3.3 --- Phylogeny of the ligninolytic enzyme-coding genes --- p.170 / Chapter 4.3.3.1 --- Laccase coding genes --- p.170 / Chapter 4.3.3.2 --- MnP coding genes --- p.175 / Chapter 4.3.4 --- Transcription of the ligninolytic enzyme-coding genes --- p.178 / Chapter 4.4 --- Treatment of DDE by 1st SMC of P. pulmonarius grown on straw-based compost --- p.183 / Chapter 4.4.1 --- Optimization of DDE removal in soil system --- p.183 / Chapter 4.5 --- Treatment of DDE by crude enzyme preparations of P. pulmonarius grown on straw-based compost --- p.184 / Chapter 4.6 --- Cost-effectiveness of the bioremediation method --- p.185 / Chapter 4.7 --- Further investigations --- p.194 / Chapter Chapter V --- Conclusions --- p.197 / References --- p.199