by Wang Pui. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2002. / Includes bibliographical references (leaves 115-128). / Abstracts in English and Chinese. / Acknowledgments --- p.i / Abstract --- p.ii / List of Figures --- p.vi / List of Tables --- p.viii / Abbreviations --- p.ix / Chapter 1. --- Introduction Pg no / Chapter 1.1 --- Ligninolytic enzyme systems --- p.1 / Chapter 1.2 --- Three main ligninolytic enzymes --- p.3 / Chapter 1.2.1 --- Lignin peroxidases (LiP) --- p.3 / Chapter 1.2.2 --- Gene structure and Amino acid sequence structure --- p.7 / Chapter 1.2.3 --- Regulation of expression --- p.8 / Chapter 1.3. --- MnP --- p.8 / Chapter 1.3.1 --- General properties --- p.8 / Chapter 1.3.2 --- Gene structure and Amino acid sequence --- p.9 / Chapter 1.3.3 --- Regulation of Expression --- p.12 / Chapter 1.4 --- Laccase --- p.12 / Chapter 1.4.1 --- General Properties --- p.12 / Chapter 1.4.2 --- Gene structure and Amino acid sequence --- p.14 / Chapter 1.5 --- Pentachlorophenol (PCP) --- p.16 / Chapter 1.5.1 --- Production --- p.16 / Chapter 1.5.2 --- Toxicity --- p.15 / Chapter 1.5.3 --- Persistence --- p.19 / Chapter 1.6 --- Oyster mushroom --- p.22 / Chapter 1.7 --- Application of ligninolytic enzymes in bioremediation --- p.23 / Chapter 1.7.1 --- Genetic modification --- p.23 / Chapter 1.7.2 --- Characterization of enzymes properties --- p.25 / Chapter 1.7.3 --- Ligninolytic enzymes Purification and extraction --- p.26 / Chapter 1.7.4 --- Immobilization of ligninolytic enzymes --- p.26 / Chapter 1.8 --- Fermentation --- p.29 / Chapter 1.8.1 --- Different types of fermentation --- p.29 / Chapter 1.8.1.1 --- Submerged fermentation (SF) --- p.29 / Chapter 1.8.1.2 --- Solid State Fermentation (SSF) --- p.30 / Chapter 1.9 --- Proposal and experimental plan of the project --- p.33 / Chapter 1.9.1 --- Objectives --- p.34 / Chapter 2. --- Methods --- p.36 / Chapter 2.1 --- Materials / Chapter 2.1.1 --- Culture maintenance --- p.36 / Chapter 2.1.2 --- Preparation of Pentachlorophenol (PCP) stock solution --- p.36 / Chapter 2.2 --- Optimization of production of ligninolytic enzymes by effective PCP concentration --- p.37 / Chapter 2.2.1 --- Preparation of mycelial homogenate --- p.37 / Chapter 2.2.2 --- Incubation --- p.37 / Chapter 2.2.3 --- Specific enzyme assays --- p.38 / Chapter 2.2.3.1 --- Laccase --- p.38 / Chapter 2.2.3.2 --- Manganese peroxidase (MnP) --- p.39 / Chapter 2.2.3.3 --- Lignin peroxidase (LiP) --- p.39 / Chapter 2.2.3.4 --- Protein --- p.39 / Chapter 2.3 --- Cloning of specific PCP-degradative laccase cDNA --- p.40 / Chapter 2.3.1 --- Isolation of total RNA --- p.41 / Chapter 2.3.2 --- Spectrophotometric quantification and qualification of DNA and RNA --- p.41 / Chapter 2.3.3 --- First strand cDNA synthesis --- p.42 / Chapter 2.3.4 --- Amplification of laccase cDNA --- p.43 / Chapter 2.3.4.1 --- Design of primers for PCR reaction --- p.43 / Chapter 2.3.4.2 --- Polymerase chain reaction --- p.44 / Chapter 2.3.5 --- Agarose gel electrophoresis of DNA --- p.44 / Chapter 2.3.6 --- Purification of PCR products --- p.45 / Chapter 2.3.7 --- TA cloning of PCR products --- p.46 / Chapter 2.3.8 --- Preparation of Escherichia coli competent cells --- p.46 / Chapter 2.3.9 --- Bacterial transformation by heat shock --- p.47 / Chapter 2.3.10 --- Colony screening --- p.48 / Chapter 2.3.11 --- Mini-preparation of plasmid DNA --- p.48 / Chapter 2.3.12 --- Sequencing --- p.49 / Chapter 2.3.13 --- Identification of sequence --- p.51 / Chapter 2.4 --- Study of regulation temporal expression of laccase genes by PCP --- p.51 / Chapter 2.4.1 --- Semi-quantitative PCR --- p.51 / Chapter 2.4.1.1 --- Design of gene-specific primers --- p.51 / Chapter 2.4.1.2 --- Determination of suitable PCR cycles --- p.54 / Chapter 2.4.1.3 --- Normalization of the amount of RNA of each sample --- p.54 / Chapter 2.5 --- Quantification of residual PCP concentration --- p.55 / Chapter 2.5.1 --- Extraction of PCP --- p.55 / Chapter 2.5.2 --- High performance liquid chromatography --- p.55 / Chapter 2.5.3 --- Assessment criteria --- p.56 / Chapter 2.6 --- Effect of other componds on laccase activity and laccase expression --- p.56 / Chapter 2.6.1 --- Study of different isoform of laccase --- p.57 / Chapter 2.6.2 --- SDS-PAGE analysis of proteins --- p.58 / Chapter 2.7 --- Study of laccase expression and laccase activity in fruiting process of oyster mushroom --- p.59 / Chapter 2.8 --- Statistical analysis --- p.60 / Chapter 3. --- Results --- p.61 / Chapter 3.1 --- Production of Ligninolytic Enzymes by oyster mushroom / Chapter 3.1.1 --- Optimization of laccase production --- p.62 / Chapter 3.1.2 --- Optimization of MnP production --- p.64 / Chapter 3.1.3 --- Change of Protein content at different PCP concentration and time --- p.64 / Chapter 3.1.4 --- Change of specific activity at different PCP concentration and time --- p.64 / Chapter 3.1.5 --- Toxicity of PCP towards mycelial growth --- p.67 / Chapter 3.1.6 --- Enzyme productivities of laccase and MnP --- p.67 / Chapter 3.1.7 --- Change of % of residual PCP concentrations during 14 days --- p.70 / Chapter 3.2. --- Cloning of PCP-degradative laccase genes --- p.70 / Chapter 3.3 --- Regulation of expression of the laccase genes by PCP --- p.74 / Chapter 3.3.1 --- Determination of suitable PCR cycles --- p.74 / Chapter 3.3.2 --- Normalization of total RNA amount of different samples --- p.74 / Chapter 3.3.3 --- Regulation of temporal expression of the laccase genes by PCP --- p.74 / Chapter 3.4 --- Effect of other compounds and physiological status on laccase activity and expression --- p.81 / Chapter 3.5 --- Study of different forms of laccase --- p.86 / Chapter 4. --- Discussion --- p.93 / Chapter 4.1 --- Production of Ligninolytic enzymes by Pleurotus pulmonarius / Chapter 4.1.1 --- Optimization of laccase and MnP production by PCP --- p.95 / Chapter 4.2 --- Cloning of laccase genes --- p.97 / Chapter 4.2.1 --- Cloning strategy --- p.97 / Chapter 4.2.2 --- Analysis of Nucleotide sequence of Lac1 - Lac3 --- p.99 / Chapter 4.2.3 --- Characterization and comparison of deduced amino acid sequences of Lacl-Lac3 --- p.99 / Chapter 4.3 --- Regulation of expression of the laccase genes by PCP --- p.100 / Chapter 4.3.1 --- Regulation of temporal expression by PCP --- p.100 / Chapter 4.4 --- Effect of the potential inducers on laccase activity and expression --- p.103 / Chapter 4.5 --- Effect of the physiological status on laccase activity and expression --- p.105 / Chapter 4.5.1 --- Production of PCP-degradative laccase by Solid-state fermentation --- p.107 / Chapter 4.5.2 --- Uses of molecular probe in bioremediation --- p.107 / Chapter 4.6 --- Different isoforms of laccase --- p.109 / Chapter 4.7 --- Conclusion --- p.112 / Chapter 4.8 --- Further studies / Chapter 4.8.1 --- Confirmation of PCP-degradation by gene product of Lac1 and Lac2 --- p.114 / Chapter 4.8.2 --- Optimization of PCP-degradative laccases production by solid-state fermentation --- p.114 / Chapter 5. --- References --- p.115

Yau Sze-nga. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 155-177). / Abstracts in English and Chinese. / Acknowledgement --- p.i / Abstract --- p.ii / 摘要 --- p.v / Table of Contents --- p.vii / List of Figures --- p.xi / List of Tables --- p.xiv / Chapter 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- Organopollutants and environment --- p.1 / Chapter 1.2 --- Pentachlorophenol --- p.3 / Chapter 1.2.1 --- Application of pentachlorophenol --- p.3 / Chapter 1.2.2 --- Characteristics of PCP --- p.4 / Chapter 1.2.3 --- Toxicity of PCP --- p.5 / Chapter 1.2.4 --- Environmental exposure of PCP --- p.6 / Chapter 1.3 --- Wastewater treatments of organopollutants --- p.9 / Chapter 1.3.1 --- Physical treatment --- p.10 / Chapter 1.3.2 --- Chemical treatment --- p.10 / Chapter 1.3.3 --- Bioremediation --- p.11 / Chapter 1.4 --- Biodegradation of PCP --- p.13 / Chapter 1.4.1 --- Biodegradation of PCP by bacteria --- p.13 / Chapter 1.4.2 --- Biodegradation of PCP by fungi --- p.14 / Chapter 1.5 --- Ligninolytic enzyme --- p.16 / Chapter 1.5.1 --- Lignin peroxidase --- p.16 / Chapter 1.5.2 --- Manganese peroxidase --- p.19 / Chapter 1.5.3 --- Laccase --- p.21 / Chapter 1.5.4 --- Biodegradation of PCP and other organopollutants by ligninolytic enzymes --- p.25 / Chapter 1.6 --- Structure and gene regulation --- p.27 / Chapter 1.6.1 --- MnP gene and structure --- p.27 / Chapter 1.6.1.1 --- Structure of MnP --- p.27 / Chapter 1.6.1.2 --- MnP gene regulation --- p.30 / Chapter 1.6.2 --- Laccase gene and structure --- p.31 / Chapter 1.6.2.1 --- Structure of laccase --- p.31 / Chapter 1.6.2.2 --- Laccase gene regulation --- p.32 / Chapter 1.7 --- Pleurotus pulmonarius --- p.36 / Chapter 1.8 --- Aims of study --- p.37 / Chapter 2 --- MATERIALS & METHOD --- p.39 / Chapter 2.1 --- Optimization of PCP induction in broth system --- p.39 / Chapter 2.1.1 --- Specific enzyme assays --- p.41 / Chapter 2.1.1.1 --- Assay for laccase activity --- p.41 / Chapter 2.1.1.2 --- Assay for manganese peroxidase (MnP) activity --- p.41 / Chapter 2.1.1.3 --- Assay for protein assay --- p.41 / Chapter 2.1.2 --- PCP effect on biomass gain --- p.42 / Chapter 2.1.3 --- Extraction of PCP --- p.42 / Chapter 2.1.3.1 --- Preparation of PCP stock solution --- p.43 / Chapter 2.1.3.2 --- Extraction efficiency of PCP --- p.43 / Chapter 2.1.3.3 --- Quantification of PCP by HPLC --- p.43 / Chapter 2.1.3.4 --- Study of PCP degradation pathway using GC-MS --- p.44 / Chapter 2.2 --- Isolation of laccase and manganese peroxidase coding genes --- p.46 / Chapter 2.2.1 --- Preparation of ribonuclease free reagents and apparatus --- p.46 / Chapter 2.2.2 --- Isolation of RNA --- p.46 / Chapter 2.2.3 --- Quantification of total RNA --- p.47 / Chapter 2.2.4 --- First strand cDNA synthesis --- p.47 / Chapter 2.2.5 --- Polymerase Chain Reaction (PCR) --- p.48 / Chapter 2.2.6 --- Gel electrophoresis --- p.50 / Chapter 2.2.7 --- Purification of PCR products --- p.50 / Chapter 2.2.8 --- Preparation of Escherichia coli competent cells --- p.51 / Chapter 2.2.9 --- Ligation and E. coli transformation --- p.51 / Chapter 2.2.10 --- PCR screening of E. coli transformation --- p.52 / Chapter 2.2.11 --- Isolation of recombinant plasmid --- p.52 / Chapter 2.2.12 --- Sequence analysis --- p.53 / Chapter 2.2.13 --- Construction of dendrogram for Pleurotus sp. laccase and manganese peroxidase dendrogram --- p.54 / Chapter 2.2.13.1 --- Dendrogram of laccase genes --- p.55 / Chapter 2.2.13.2 --- Dendrogram of manganese genes --- p.55 / Chapter 2.3 --- Differential regulation profiles of laccase and manganese peroxidase genes --- p.57 / Chapter 2.3.1 --- Time course of the effects of PCP on levels of laccase and manganese peroxidase mRNAs --- p.57 / Chapter 2.3.1.1 --- Isolation of RNA --- p.57 / Chapter 2.3.1.2 --- RT-PCR --- p.57 / Chapter 2.3.2 --- The effect of different stresses --- p.65 / Chapter 2.3.2.1 --- Pollutant removal analysis --- p.66 / Chapter 2.3.2.2 --- Differential gene expression under different stresses --- p.69 / Chapter 2.4 --- Construction of full-length cDNA --- p.69 / Chapter 2.4.1 --- Primer design --- p.69 / Chapter 2.4.2 --- First-strand cDNA synthesis --- p.71 / Chapter 2.4.3 --- RACE PCR reactions --- p.71 / Chapter 2.5 --- Statistical analysis --- p.73 / Chapter 3 --- RESULT --- p.74 / Chapter 3.1 --- Optimization of PCP induction in broth system --- p.74 / Chapter 3.1.1 --- Enzyme Assay --- p.74 / Chapter 3.1.1.1 --- Protein content --- p.74 / Chapter 3.1.1.2 --- Specific laccase activity --- p.74 / Chapter 3.1.1.3 --- Specific MnP activity --- p.76 / Chapter 3.1.1.4 --- Laccase productivity --- p.78 / Chapter 3.1.1.5 --- MnP productivity --- p.78 / Chapter 3.1.2 --- PCP effect on biomass development --- p.80 / Chapter 3.1.3 --- PCP removal --- p.80 / Chapter 3.2 --- isolation of laccase and manganese peroxidase coding genes --- p.83 / Chapter 3.2.1 --- Dendrogram construction for heterologous MnP and laccase coding genes --- p.83 / Chapter 3.2.2 --- Phylogeny of ligninolytic enzyme coding genes of P. pulmonarius --- p.85 / Chapter 3.2.2.1 --- Phylogeny of MnP coding genes --- p.88 / Chapter 3.2.2.2 --- Phylogeny of laccase coding genes --- p.88 / Chapter 3.3 --- differential regulation profiles of laccase and MnP genes --- p.91 / Chapter 3.3.1 --- Time course of the effects of PCP on levels of MnP and laccase mRNAs --- p.91 / Chapter 3.3.1.1 --- Time course of the effects of PCP on levels of MnP mRNAs --- p.91 / Chapter 3.3.1.2 --- Time course of the effects of PCP on levels of laccase mRNAs --- p.97 / Chapter 3.3.2 --- The effects of different stresses and two lignocellulosic substrates --- p.99 / Chapter 3.3.2.1 --- The effect on laccase and MnP enzyme activities --- p.99 / Chapter 3.3.2.1.1 --- Protein content --- p.99 / Chapter 3.3.2.1.2 --- Specific laccase activity --- p.100 / Chapter 3.3.2.1.3 --- Specific MnP activity --- p.102 / Chapter 3.3.2.1.4 --- Dry weight of P. pulmonarius --- p.102 / Chapter 3.3.2.1.5 --- Laccase productivity --- p.105 / Chapter 3.3.2.1.6 --- MnP productivity --- p.105 / Chapter 3.3.2.2 --- Organopollutant removal --- p.107 / Chapter 3.3.2.3 --- Differential gene expression under different stresses --- p.107 / Chapter 3.3.2.3.1 --- The effect on MnP mRNAs --- p.107 / Chapter 3.3.2.3.2 --- The effect on laccase mRNAs --- p.115 / Chapter 3.4 --- Construction of full-length cDNA --- p.116 / Chapter 3.4.1 --- PPMnP5 --- p.117 / Chapter 3.4.2 --- PPlac2 --- p.120 / Chapter 3.4.3 --- PPlac6 --- p.120 / Chapter 4 --- DISCUSSION --- p.123 / Chapter 4.1 --- Optimization of PCP induction in broth system --- p.123 / Chapter 4.2 --- Isolation of MnP and laccase coding genes --- p.126 / Chapter 4.3 --- Differential regulation profiles of MnP and laccase genes --- p.128 / Chapter 4.3.1 --- The effects incubation time and PCP on levels of MnP and laccase mRNAs --- p.128 / Chapter 4.3.1.1 --- MnP --- p.129 / Chapter 4.3.1.2 --- Laccase --- p.129 / Chapter 4.3.2 --- Regulation of MnP and laccase by different substrates --- p.130 / Chapter 4.3.2.1 --- Regulation of MnP and laccase activities --- p.131 / Chapter 4.3.2.2 --- Organopollutant removal --- p.132 / Chapter 4.3.2.3 --- Regulation of MnP coding genes --- p.136 / Chapter 4.3.2.4 --- Regulation of laccase coding genes --- p.137 / Chapter 4.4 --- "Characterization of full length cDNAs of PPMnP5, PPlac2 and PPLAC6" --- p.140 / Chapter 4.4.1 --- PPMnP5 --- p.140 / Chapter 4.4.2 --- PPlac2 and PPlac6 --- p.144 / Chapter 4.4.3 --- Real-time PCR --- p.146 / Chapter 4.4.3.1 --- Methodology for SYBR-Green real-time PCR --- p.146 / Chapter 4.4.3.2 --- Comparison of conventional PCR and real-time PCR --- p.148 / Chapter 4.5 --- APPLICATION AND FURTHER INVESTIGATION --- p.150 / Chapter 5 --- CONCLUSION --- p.152 / Chapter 6 --- REFERENCES --- p.155

by Wai Lok Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 142-155). / Abstracts in English and Chinese. / Acknowledgements --- p.i / Abstracts --- p.ii / Contents --- p.vii / List of figures --- p.xiii / List of tables --- p.xvi / Abbreviations --- p.xviii / Chapter 1. --- Introduction --- p.1 / Chapter 1.1 --- Pentachlorophenol / Chapter 1.1.1 --- Applications of pentachlorophenol --- p.1 / Chapter 1.1.2 --- Characteristics --- p.3 / Chapter 1.1.3 --- Pentachlorophenol in the environment --- p.3 / Chapter 1.1.4 --- Toxicity of Pentachlorophenol --- p.6 / Chapter 1.2 --- Treatments of Pentachlorophenol --- p.10 / Chapter 1.2.1 --- Physical treatment --- p.10 / Chapter 1.2.2 --- Chemical treatment --- p.11 / Chapter 1.2.3 --- Biological treatment --- p.13 / Chapter 1.3 --- Biodegradation --- p.14 / Chapter 1.3.1 --- Biodegradation of PCP by bacteria --- p.14 / Chapter 1.3.2 --- Biodegradation of PCP by white-rot fungi --- p.15 / Chapter 1.4 --- Biosorption --- p.24 / Chapter 1.5 --- Proposed Strategy --- p.28 / Chapter 1.6 --- Spent Mushroom Compost / Chapter 1.6.1 --- Background --- p.28 / Chapter 1.6.2 --- Physico-chemical properties of SMC --- p.29 / Chapter 1.6.3 --- As a biosorbent --- p.29 / Chapter 1.6.3.1 --- Factors affecting biosorption --- p.31 / Chapter 1.6.3.2 --- Contact time --- p.31 / Chapter 1.6.3.3 --- Initial pH --- p.32 / Chapter 1.6.3.4 --- Concentration of biosorbent --- p.33 / Chapter 1.6.3.5 --- Initial PCP concentration --- p.34 / Chapter 1.6.3.6 --- Incubation temperature --- p.34 / Chapter 1.6.3.7 --- Agitation speed --- p.35 / Chapter 1.6.4 --- Modeling of adsorption --- p.36 / Chapter 1.6.4.1 --- Langmuir isotherm --- p.36 / Chapter 1.6.4.2 --- Freundlich isotherm --- p.36 / Chapter 1.6.5 --- As a source of PCP-degrading bacteria --- p.38 / Chapter 1.6.5.1 --- Identification of PCP-degrading bacterium --- p.40 / Chapter 1.6.6 --- As a source of fungus --- p.42 / Chapter 1.7 --- Objectives of this Study --- p.43 / Chapter 2. --- Materials and Methods --- p.44 / Chapter 2.1 --- Spent Mushroom compost (SMC) Production --- p.44 / Chapter 2.2 --- Characterization of SMC --- p.46 / Chapter 2.2.1 --- pH --- p.46 / Chapter 2.2.2 --- Electrical conductivity --- p.46 / Chapter 2.2.3 --- "Carbon, hydrogen, nitrogen and sulphur contents" --- p.46 / Chapter 2.2.4 --- Infrared spectroscopic study --- p.47 / Chapter 2.2.5 --- Metal analysis --- p.47 / Chapter 2.2.6 --- Anion content --- p.47 / Chapter 2.2.7. --- Chitin assay --- p.48 / Chapter 2.3 --- Extraction of PCP --- p.49 / Chapter 2.3.1 --- Selection of extraction solvent --- p.49 / Chapter 2.3.2 --- Selection of desorbing agent --- p.49 / Chapter 2.3.3 --- Extraction efficiency --- p.50 / Chapter 2.4 --- Adsorption of Pentachlorophenol on SMC --- p.50 / Chapter 2.4.1 --- Preparation of pentachlorophenol (PCP) stock solution --- p.50 / Chapter 2.4.2 --- Batch adsorption experiment --- p.51 / Chapter 2.4.3 --- Quantification of PCP by HPLC --- p.51 / Chapter 2.4.4 --- Data analysis for biosorption --- p.51 / Chapter 2.4.5 --- Optimization of PCP adsorption --- p.52 / Chapter 2.4.5.1 --- Effect of contact time --- p.52 / Chapter 2.4.5.2 --- Effect of initial pH --- p.52 / Chapter 2.4.5.3 --- Effect of incubation temperature --- p.53 / Chapter 2.4.5.4 --- Effect of shaking speed --- p.53 / Chapter 2.4.5.5 --- Effect of initial PCP concentration and amount of biosorbent --- p.53 / Chapter 2.4.6 --- Adsorption isotherm --- p.53 / Chapter 2.4.7 --- Effect of removal efficiency on reuse of biosorbent --- p.54 / Chapter 2.5 --- Biodegradation by Isolated Bacterium --- p.54 / Chapter 2.5.1 --- Isolation of PCP-tolerant bacteria from mushroom compost --- p.54 / Chapter 2.5.2 --- Screening for the best PCP-tolerant bacterium --- p.54 / Chapter 2.5.3 --- Identification of the isolated bacterium --- p.55 / Chapter 2.5.3.1 --- 16S ribosomal DNA sequencing --- p.55 / Chapter 2.5.3.1.1 --- Extraction of DNA --- p.55 / Chapter 2.5.3.1.2 --- Specific PCR for 16S rDNA --- p.56 / Chapter 2.5.3.1.3 --- Gel electrophoresis --- p.57 / Chapter 2.5.3.1.4 --- Purification of PCR products --- p.57 / Chapter 2.5.3.1.5 --- Sequencing of 16S rDNA --- p.58 / Chapter 2.5.3.2 --- Gram staining --- p.60 / Chapter 2.5.3.3 --- Biolog Microstation System --- p.60 / Chapter 2.5.3.4 --- MIDI Sherlock Microbial Identification System --- p.61 / Chapter 2.5.4 --- Optimization of PCP degradation by PCP-degrading bacterium --- p.62 / Chapter 2.5.4.1 --- Effect of incubation time --- p.63 / Chapter 2.5.4.2 --- Effect of shaking speed --- p.63 / Chapter 2.5.4.3 --- Effect of initial PCP concentration and inoculum size --- p.63 / Chapter 2.5.4.4 --- Study of PCP degradation pathway by isolated bacterium using GC-MS --- p.64 / Chapter 2.6 --- Biodegradation by Fungus Pleurotus pulmonarius --- p.64 / Chapter 2.6.1 --- Optimization of PCP degradation by P. pulmonarius --- p.65 / Chapter 2.6.1.1 --- Effect of incubation time --- p.65 / Chapter 2.6.1.2 --- Effect of shaking speed --- p.65 / Chapter 2.6.1.3 --- Effect of initial PCP concentration and inoculum size --- p.65 / Chapter 2.6.2 --- Study of PCP degradation pathway by fungus using GC-MS --- p.65 / Chapter 2.6.3 --- Specific enzyme assays --- p.66 / Chapter 2.6.3.1 --- Extraction of protein and enzymes --- p.66 / Chapter 2.6.3.2 --- Protein --- p.66 / Chapter 2.6.3.3 --- Laccase --- p.67 / Chapter 2.6.3.4 --- Manganese peroxidase (MnP) --- p.67 / Chapter 2.6.4 --- Microtox® assay --- p.67 / Chapter 2.7 --- Statistical Analysis --- p.68 / Chapter 3. --- Results --- p.69 / Chapter 3.1 --- Physico-chemical Properties of SMC --- p.69 / Chapter 3.2 --- Extraction Efficiency and Desorption Efficiency of PCP --- p.69 / Chapter 3.3 --- Batch Adsorption Experiments --- p.76 / Chapter 3.3.1 --- Optimization of adsorption conditions --- p.76 / Chapter 3.3.1.1 --- Effect of contact time --- p.76 / Chapter 3.3.1.2 --- Effect of initial pH --- p.76 / Chapter 3.3.1.3 --- Effect of shaking speed --- p.79 / Chapter 3.3.1.4 --- Effect of incubation temperature --- p.79 / Chapter 3.3.1.5 --- Effect of initial PCP concentration and amount of biosorbent --- p.79 / Chapter 3.3.2 --- Reuse of SMC --- p.83 / Chapter 3.3.3 --- Isotherm plot --- p.83 / Chapter 3.4 --- Biodegradation by PCP-degrading Bacterium --- p.86 / Chapter 3.4.1 --- Isolation and purification of PCP-tolerant bacteria --- p.86 / Chapter 3.4.2 --- Identification of the isolated bacterium --- p.90 / Chapter 3.4.2.1 --- 16S rDNA sequencing --- p.90 / Chapter 3.4.2.2 --- Gram staining --- p.90 / Chapter 3.4.2.3 --- Biolog MicroPlates Identification System --- p.90 / Chapter 3.4.2.4 --- MIDI Sherlock Microbial Identification System --- p.90 / Chapter 3.4.3 --- Growth curve of PCP-degrading bacterium --- p.90 / Chapter 3.4.4 --- Optimization of PCP degradation by PCP-degrading bacterium --- p.97 / Chapter 3.4.4.1 --- Effect of incubation time --- p.97 / Chapter 3.4.4.2 --- Effect of shaking speed --- p.97 / Chapter 3.4.4.3 --- Effect of initial PCP concentration and inoculum size of bacterium --- p.101 / Chapter 3.4.5 --- Determination of breakdown products of PCP by PCP-degrading bacterium --- p.101 / Chapter 3.5 --- Biodegradation by Fungus Pleurotus pulmonarius --- p.103 / Chapter 3.5.1 --- Growth curve of P. pulmonarius --- p.103 / Chapter 3.5.2 --- Optimization of PCP degradation by P. pulmonarius --- p.103 / Chapter 3.5.2.1 --- Effect of incubation time --- p.103 / Chapter 3.5.2.2 --- Effect of shaking speed --- p.103 / Chapter 3.5.2.3 --- Effect of initial PCP concentration and inoculum size of fungus --- p.108 / Chapter 3.5.3 --- Determination of breakdown products of PCP by P. pulmonarius --- p.108 / Chapter 3.5.4 --- Enzyme assays --- p.108 / Chapter 3.6 --- Integration of Biosorption by SMC and Biodegradation by P. pulmonarius --- p.112 / Chapter 3.6.1 --- Evaluation of PCP removal by an integration system --- p.112 / Chapter 3.6.2 --- Evaluation of toxicity by Micortox® assays --- p.112 / Chapter 4. --- Discussion --- p.115 / Chapter 4.1 --- Physico-chemical Properties of SMC --- p.115 / Chapter 4.2 --- Extraction Efficiency and Desorption Efficiency of PCP --- p.116 / Chapter 4.3 --- Batch Biosorption Experiment --- p.117 / Chapter 4.3.1 --- Effect of contact time --- p.117 / Chapter 4.3.2 --- Effect of initial pH --- p.118 / Chapter 4.3.3 --- Effect of shaking speed --- p.120 / Chapter 4.3.4 --- Effect of incubation temperature --- p.120 / Chapter 4.3.5 --- Effect of initial PCP concentration and amount of biosorbent --- p.121 / Chapter 4.3.6 --- Reuse of SMC --- p.122 / Chapter 4.3.7 --- Modeling of biosorption --- p.122 / Chapter 4.4 --- Biodegradation of PCP by PCP-degrading Bacterium --- p.124 / Chapter 4.4.1 --- Isolation and purification of PCP-tolerant bacterium --- p.124 / Chapter 4.4.2 --- Identification of the isolated bacterium --- p.125 / Chapter 4.4.3 --- Optimization of PCP degradation by PCP-degrading bacterium --- p.126 / Chapter 4.4.3.1 --- Effect of incubation time --- p.126 / Chapter 4.4.3.2 --- Effect of shaking speed --- p.128 / Chapter 4.4.3.3 --- Effect of initial PCP concentration and inoculum size of bacterium --- p.128 / Chapter 4.4.4 --- PCP degradation pathway by S. marcescens --- p.129 / Chapter 4.5 --- Biodegradation of PCP by Pleurotus pulmonarius --- p.130 / Chapter 4.5.1 --- Optimization of PCP degradation by P. pulmonarius --- p.130 / Chapter 4.5.1.1 --- Effect of incubation time --- p.131 / Chapter 4.5.1.2 --- Effect of shaking speed --- p.131 / Chapter 4.5.1.3 --- Effect of initial PCP concentration and inoculum size of fungus --- p.131 / Chapter 4.5.2 --- Enzyme activities --- p.132 / Chapter 4.5.3 --- PCP degradation pathway by P. pulmonarius --- p.133 / Chapter 4.6 --- Comparison of PCP Degradation between S.marcescens and P. pulmonarius --- p.133 / Chapter 4.7 --- Integration of Biosorption by SMC and Biodegradation by P. pulmonarius --- p.135 / Chapter 4.8 --- Evaluation of toxicity by Microtox® assay --- p.135 / Chapter 4.9 --- Comparison of PCP Removal by Integration System of Sorption and Fungal Biodegradation and Conventional Treatments --- p.136 / Chapter 4.10 --- Further Investigations --- p.137 / Chapter 5. --- Conclusion --- p.139 / Chapter 6. --- References --- p.142

Organochlorine (OC) compounds that include several pesticides as well as an array of industrial chemicals were very efficacious for their intended use but were also characterized by deleterious environmental impacts when released either intentionally or inadvertently. Their lipophilic nature, long persistence in the environment and threat to human health caused all the developed countries to ban the production of these chemicals as well as restricted the use of formulations containing these material for food production. / A number of scientists have become involved in the development of intentional degradation methods/techniques for these compounds using zero-valent metals or bimetallic mixtures. To date, there is no single, simple and continuous procedure available to completely dechlorinate lindane or pentachlorophenol (PCP). This work describes the complete dechlorination of lindane and pentachlorophenol by zero-valent Zn, Fe and Fe/Ag bimetallic mixture as well as a supercritical fluid extraction technique for a more efficient mass transfer of the substrates to the surfaces of the metal catalyst. The dechlorination reaction occurs on the surface of metal particles with the removal of all the chlorine atoms from lindane and PCP in a matter of minute, and yields completely dechlorinated hydrocarbon molecules and chloride as products. (Abstract shortened by UMI.)

No description available.

Dominant genera of bacteria were isolated from three river waters during anthracene and pentachlorophenol biotransformation studies. The genera Pseudomonas, Acinetobacter, Micrococcus, Chromobacterium, Alcaligenes, Azomonos, Bacillus, and Flavobacterium were capable of biotransforming one or both of these compounds. These isolates were subjected to further biotransformation tests, including river water and a basal salt medium with and without additional glucose. The results of these experiments were evaluated statistically. It was concluded that only a limited number of the bacteria identified were able to transform these chemicals in river water. The addition of glucose to the growth medium significantly affected the biotransformation of these chemicals. It was also determined that the size of the initial bacterial population is not a factor in determining whether biotransformation of anthracene or pentachlorophenol can occur.

by Law Wing Man. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 192-206). / Abstracts in English and Chinese. / Acknowledgments --- p.i / Abstract --- p.ii / List of Figures --- p.vi / List of Tables --- p.xii / Abbreviations --- p.xv / Chapter 1. --- Introduction / Chapter 1.1. --- Pesticides --- p.1 / Chapter 1.1.1. --- Types and uses --- p.1 / Chapter 1.1.2. --- Development of pesticides --- p.1 / Chapter 1.1.3. --- The case against pesticides --- p.3 / Chapter 1.2. --- Pentachlorophenol --- p.4 / Chapter 1.2.1. --- Production --- p.4 / Chapter 1.2.2. --- Toxicity --- p.4 / Chapter 1.2.3. --- Persistency --- p.6 / Chapter 1.3. --- Methyl-parathion --- p.9 / Chapter 1.3.1. --- Production --- p.9 / Chapter 1.3.2. --- Toxicity --- p.9 / Chapter 1.3.3. --- Environmental fate --- p.12 / Chapter 1.4. --- Conventional methods dealing with pesticides --- p.12 / Chapter 1.5. --- Bioremediation --- p.15 / Chapter 1.6. --- Spent mushroom compost --- p.17 / Chapter 1.6.1. --- Background --- p.17 / Chapter 1.6.2. --- "Physical, chemical and biological properties of SMC " --- p.19 / Chapter 1.6.3. --- Recycling of agricultural residuals --- p.21 / Chapter 1.6.3.1. --- Definition --- p.21 / Chapter 1.6.3.2. --- Types of recycling --- p.22 / Chapter 1.6.4. --- Potential uses of SMC as bioremediating agent --- p.23 / Chapter 1.6.4.1. --- Use of microorganisms in SMC --- p.23 / Chapter 1.6.4.2. --- Use of ligninolytic enzymes in SMC --- p.24 / Chapter 1.7. --- Ligninolytic enzymes --- p.28 / Chapter 1.7.1. --- Background --- p.28 / Chapter 1.7.2. --- What are white rot fungi? --- p.29 / Chapter 1.7.3. --- Why is lignin so difficult to degrade? --- p.29 / Chapter 1.7.4. --- Three main ligninolytic enzymes --- p.32 / Chapter 1.7.4.1. --- Lignin peroxidases (LiP) --- p.32 / Chapter 1.7.4.2. --- Manganese peroxidase (MnP) --- p.36 / Chapter 1.7.4.3. --- Laccase --- p.37 / Chapter 1.8. --- Why SMC was chosen to be the bioremediating agent in my project? --- p.40 / Chapter 1.9. --- Bioremediation of chlorophenols and PCP --- p.44 / Chapter 1.9.1. --- Bacterial system --- p.44 / Chapter 1.9.2. --- Fungal system --- p.45 / Chapter 1.10. --- Bioremediation of methyl-parathion --- p.49 / Chapter 1.10.1. --- Bacterial system --- p.49 / Chapter 1.10.2. --- Fungal system --- p.51 / Chapter 1.11. --- Proposal and experimental plan of the project --- p.51 / Chapter 1.11.1. --- Study the removal of pesticides in both aquatic and soil system --- p.52 / Chapter 1.11.2. --- Research strategy --- p.52 / Chapter 1.11.3. --- Optimization of pesticide removal --- p.53 / Chapter 1.11.4. --- Identification of breakdown products --- p.54 / Chapter 1.11.5. --- Toxicity assay --- p.54 / Chapter 1.11.6. --- Isotherm plot --- p.55 / Chapter 1.12. --- Objectives of the study --- p.56 / Chapter 2. --- Material and Methods --- p.58 / Chapter 2.1. --- Material --- p.59 / Chapter 2.2. --- Production of Spent Mushroom Compost (SMC) --- p.59 / Chapter 2.3. --- Characterization of SMC --- p.60 / Chapter 2.3.1. --- PH --- p.60 / Chapter 2.3.2. --- Electrical conductivity --- p.60 / Chapter 2.3.3. --- "Carbon, hydrogen, nitrogen and sulphur contents " --- p.60 / Chapter 2.3.4. --- Ash content --- p.61 / Chapter 2.3.5. --- Metal analysis --- p.61 / Chapter 2.3.6. --- Anion content --- p.62 / Chapter 2.3.7. --- Chitin assay --- p.62 / Chapter 2.4. --- Characterization of soil --- p.63 / Chapter 2.4.1. --- Soil texture --- p.63 / Chapter 2.4.2. --- Moisture content --- p.64 / Chapter 2.5. --- Basic studies on the removal capacity of pesticides by SMC --- p.65 / Chapter 2.5.1. --- Preparation of pentachlorophenol and methyl- parathion stock solution --- p.66 / Chapter 2.6. --- Experimental design --- p.65 / Chapter 2.6.1. --- In aquatic system --- p.65 / Chapter 2.6.2. --- In soil system --- p.68 / Chapter 2.7. --- Extraction of pesticides --- p.68 / Chapter 2.7.1. --- In aquatic system --- p.68 / Chapter 2.7.2. --- In soil system --- p.69 / Chapter 2.8. --- Quantification of pesticides --- p.69 / Chapter 2.8.1. --- By high performance liquid chromatography --- p.69 / Chapter 2.8.2. --- By gas chromatography-mass spectrometry --- p.71 / Chapter 2.9. --- Optimization of pesticides degradation by SMC in both aquatic and soil systems --- p.72 / Chapter 2.9.1. --- Effect of initial pesticide concentrations on the removal of pesticides --- p.72 / Chapter 2.9.2. --- Effect of amount of SMC used on the removal of pesticides --- p.73 / Chapter 2.9.3. --- Effect of incubatoin time on the removal of pesticides --- p.73 / Chapter 2.9.4. --- Effect of initial pH on the removal of pesticides --- p.73 / Chapter 2.9.5. --- Effect of incubation of temperature on the removal of pesticides --- p.74 / Chapter 2.10. --- The study of breakdown process of pesticides --- p.74 / Chapter 2.10.1. --- GC/MS --- p.74 / Chapter 2.10.2. --- Ion chmatography --- p.74 / Chapter 2.11. --- Microtox® assay --- p.75 / Chapter 2.12. --- Assessment criteria --- p.75 / Chapter 2.12.1. --- In aquatic system --- p.75 / Chapter 2.12.2. --- In soil system --- p.76 / Chapter 2.13. --- Statistical analysis --- p.77 / Chapter 3. --- Results / Chapter 3.1. --- Characterization of SMC and soil --- p.78 / Chapter 3.2. --- Quantification of pesticides by HPLC and GC/MS --- p.82 / Chapter 3.3. --- Extraction efficiencies of pesticides with hexane --- p.82 / Chapter 3.4. --- Stability of pesticides against time --- p.82 / Chapter 3.5. --- Effect of sterilization of soil in the removal abilities of pesticides…… --- p.88 / Chapter 3.6. --- Optimization of removal of pentachlorophnol --- p.88 / Chapter 3.6.1. --- Effect of incubation time --- p.88 / Chapter 3.6.1.1. --- In aquatic system --- p.88 / Chapter 3.6.1.2. --- In soil system --- p.88 / Chapter 3.6.2. --- Effect of initial PCP concentrations and amout of SMC used --- p.91 / Chapter 3.6.2.1. --- In aquatic system --- p.91 / Chapter 3.6.2.2. --- In soil system --- p.94 / Chapter 3.6.3. --- Effect of pH --- p.97 / Chapter 3.6.3.1. --- In aquatic system --- p.97 / Chapter 3.6.3.2. --- In soil system --- p.97 / Chapter 3.6.4. --- Effect of incubation temperature --- p.97 / Chapter 3.6.4.1. --- In aquatic system --- p.97 / Chapter 3.6.4.2. --- In soil system --- p.101 / Chapter 3.6.5. --- Potential breakdown intermediates and products --- p.101 / Chapter 3.6.5.1. --- In aquatic system --- p.101 / Chapter 3.6.5.2. --- In soil system --- p.104 / Chapter 3.7. --- Microtox® assay of PCP --- p.110 / Chapter 3.7.1. --- In aquatic system --- p.110 / Chapter 3.7.2. --- In soil system --- p.110 / Chapter 3.8. --- Optimization of removal of methyl-parathion --- p.113 / Chapter 3.8.1. --- Effect of incubation time --- p.113 / Chapter 3.8.1.1. --- In aquatic system --- p.113 / Chapter 3.8.1.2. --- In soil system --- p.113 / Chapter 3.8.2. --- Effect of initial concentration and amount of SMC --- p.115 / Chapter 3.8.2.1. --- In aquatic system --- p.115 / Chapter 3.8.2.2. --- In soil system --- p.117 / Chapter 3.8.3. --- Effect of incubation temperature --- p.120 / Chapter 3.8.3.1. --- In aquatic system --- p.120 / Chapter 3.8.3.2. --- In soil system --- p.120 / Chapter 3.8.4. --- Potential breakdown intermediates and products --- p.121 / Chapter 3.8.4.1. --- In aquatic system --- p.121 / Chapter 3.8.4.2. --- In soil system --- p.124 / Chapter 3.9. --- Microtox ® assay of methyl-parathion --- p.133 / Chapter 3.9.1. --- In aquatic system --- p.133 / Chapter 3.9.2. --- In soil system --- p.133 / Chapter 4. --- Discussion / Chapter 4.1. --- Characterization of SMC and soil --- p.137 / Chapter 4.2. --- Stability of pesticides against time in aquatic and soil system --- p.141 / Chapter 4.3. --- Effect of sterilization of soil in the removal abilities of pesticides --- p.142 / Chapter 4.4. --- Optimization of removal of PCP --- p.142 / Chapter 4.4.1. --- Effect of incubation time --- p.142 / Chapter 4.4.1.1. --- In aquatic system --- p.142 / Chapter 4.4.1.2. --- In soil system --- p.143 / Chapter 4.4.2. --- Effect of initial PCP concentrations and amount of SMC --- p.144 / Chapter 4.4.2.1. --- In aquatic system --- p.144 / Chapter 4.4.2.2. --- In soil system --- p.147 / Chapter 4.4.3. --- Effect of pH --- p.149 / Chapter 4.4.3.1. --- In aquatic system --- p.149 / Chapter 4.4.3.2. --- In soil system --- p.150 / Chapter 4.4.4. --- Effect of incubation temperature --- p.150 / Chapter 4.4.4.1. --- In aquatic system --- p.150 / Chapter 4.4.4.2. --- In soil system --- p.152 / Chapter 4.4.5. --- Potential breakdown intermediates and products --- p.152 / Chapter 4.4.5.1. --- In aquatic system --- p.152 / Chapter 4.4.5.2. --- In soil system --- p.158 / Chapter 4.5. --- Microtox® assay of PCP --- p.159 / Chapter 4.5.1. --- In aquatic system --- p.159 / Chapter 4.5.2. --- In soil system --- p.160 / Chapter 4.6. --- Removal of PCP by the aqueous extract of SMC --- p.162 / Chapter 4.7. --- Optimization of removal of methyl-parathion --- p.164 / Chapter 4.7.1. --- Effect of incubation time --- p.164 / Chapter 4.7.1.1. --- In aquatic system --- p.164 / Chapter 4.7.1.2. --- In soil system --- p.165 / Chapter 4.7.2. --- Effect of initial methyl-paration concentrations and amount of SMC used --- p.165 / Chapter 4.7.2.1. --- In aquatic system --- p.165 / Chapter 4.7.2.2. --- I in soil system --- p.166 / Chapter 4.7.3. --- Effect of incubation temperature --- p.168 / Chapter 4.7.3.1. --- In aquatic system --- p.168 / Chapter 4.7.3.2. --- In soil system --- p.169 / Chapter 4.7.4. --- Potential breakdown intermediates and products --- p.169 / Chapter 4.7.4.1. --- In aquatic system --- p.169 / Chapter 4.7.4.2. --- In soil system --- p.170 / Chapter 4.8. --- Microtox® assay of Methyl-parathion --- p.173 / Chapter 4.8.1. --- In aquatic system --- p.173 / Chapter 4.8.2. --- In soil system --- p.174 / Chapter 4.9. --- Removal of methyl-parathion by the aqueous extract of SMC --- p.174 / Chapter 4.10. --- The ability of different types of SMC in the removal of organic pollutants --- p.176 / Chapter 4.11. --- The storage of SMC --- p.178 / Chapter 4.12. --- The effect of scale in the removal of pesticides --- p.180 / Chapter 4.13. --- Cost-effectiveness of using SMC as crude enzymes sources --- p.180 / Chapter 4.14. --- The effect of surfactant on the removal of PCP --- p.182 / Chapter 4.15. --- Prospects for employment SMC in removal of pollutants --- p.185 / Chapter 5. --- Conclusions --- p.186 / Chapter 6. --- Future investigation --- p.190 / Chapter 7. --- References --- p.192

Chlorinated aromatic compounds are an important group of compounds. Many of them have been produced in large quantities and they are indispensable to technological and societal benefits. But regulatory agencies have tightened regulations on the use and release of chlorinated aromatic compounds because of the scientific understanding of their toxicity, persistence, behavior in the environment and their potential to cause adverse effects on the ecosystem and human health. / Pentachlorophenol (PCP), octachloronaphthalene and decachlorobiphenyl are all highly chlorinated aromatic compounds, of which, PCP has been used mainly as a biocide. Octachloronaphthalene and decachlorobiphenyl don't have practical use, but their congeners have been used widely as chemicals in industry. These compounds are toxic, recalcitrant and bio-accumulated within organisms. As the conventional treatment, incineration of these compounds can cause more serious problems, so that suitable alternatives need to be developed for their detoxification. / When compared with biodegradation or the thermal treatment of these compounds, chemical degradations have several merits. (Abstract shortened by UMI.)

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