Laccases and other ligninolytic enzymes of the basidiomycetes Coprinopsis cinerea and Pleurotus ostreatus: submerged and solid state fermentation, morphological studies of liquid cultures and characterisation of new laccases
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1. Verfasser: | |
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Format: | Abschlussarbeit Buch |
Sprache: | English |
Veröffentlicht: |
Göttingen
Cuvillier Verl.
2010
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Ausgabe: | 1. Aufl. |
Schlagworte: | |
Online-Zugang: | Inhaltstext Inhaltsverzeichnis |
Beschreibung: | XII, 275 S. Ill., graph. Darst. |
ISBN: | 9783869552392 |
Internformat
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IMAGE 1
TABLE OF CONTENTS
SUMMARY VII
ZUSAMMENFASSUNG IX
ABBREVIATIONS XII
CHAPTER 1
INTRODUCTION 1
I. LACCASES: SOURCES AND APPLICATIONS 3
A. OCCURRENCE AND FUNCTIONS OF LACCASES IN NATURE 4
B. SCIENTIFIC AND COMMERCIAL APPLICATIONS 9
1. LIGNIN MODIFICATION: DEGRADATION AND POLYMERISATION 9 A) LACCASES AND
LACCASE MEDIATOR SYSTEMS (LMS) IN PULP PROCESSING 9 B) LACCASE IN THE
WOOD COMPOSITE INDUSTRY 11
C) ALTERATION OF WOOD PROPERTIES WITH LACCASES 12
2. BIOREMEDIATION 12
3. IMMOBILISATION OF LACCASE FOR DIFFERENT APPLICATIONS 15
4. COMMERCIAL LACCASE PRODUCTS 16
5. OTHER POTENTIAL APPLICATIONS 18
II. BASIDIOMYCETES: HIGHER FUNGI WITH A GREAT POTENTIAL 19
A. APPLICATIONS OF LIVING BASIDIOMYCETES ON LIGNOCELLULOSIC MATERIAL 19
B. IN VIVO BIOREMEDIATION AND DYE DECOLOURISATION 20
C. MUSHROOMS AS A VALUABLE FOOD 21
III. AIM OF THE THESIS 26
IV. REFERENCES 28
BIBLIOGRAFISCHE INFORMATIONEN HTTP://D-NB.INFO/999978500
DIGITALISIERT DURCH
IMAGE 2
CHAPTER -
LACCASES AND OTHER LIGNINOLYTIC ENZYMES OF BASIDIOMYCETES 45
I. PRODUCTION OF LACCASE AND OTHER ENZYMES FOR THE WOOD INDUSTRY 47
II. LIGNINOLYTIC ENZYME ACTIVITIES ALTERNATE WITH MUSHROOM PRODUCTION
DURING INDUSTRIAL CULTIVATION OF PLEUROTUS OSTREATUS ON WHEAT-STRAW-
BASED SUBSTRATE 87
CHAPTER 3
MORPHOLOGY OF COPRINOPSIS CINEREA IN SUBMERGED CULTURES 103 I.
INTRODUCTION: FERMENTATION OF BASIDIOMYCETES IN LIQUID CULTURES 105 A.
IMPORTANT PARAMETERS OF FERMENTATION ON FUNGAL MORPHOLOGY AND ON
PRODUCTION YIELDS OF ENZYMES AND OTHER METABOLITES 107
1. INOCULUM 107
2. OXYGEN 108
3. VESSEL AND IMPELLER 110
4. MODE OF CULTIVATION 112
B. CONCLUSION 113
II. TOOLS FOR ANALYSIS OF BIOMASS AND MORPHOLOGY OF ORGANISMS 114
A. GENERAL BIOMASS DETERMINATION 114
B. APPROACHES TO DETERMINE MORPHOLOGICAL CHARACTERISTICS OF FREE HYPHAE
AND MYCELIAL PELLETS 115
1. GENERAL IMAGE ANALYSIS 117
2. AUTOMATED IMAGE ANALYSIS 117
3. METHODS FOR A MORE PRECISE IMAGE ANALYSIS 118
4. CONCLUSION 121
III. AUTOMATED IMAGE ANALYSIS TO OBSERVE PELLET MORPHOLOGY IN LIQUID
CULTURES OF FILAMENTOUS FUNGI SUCH AS THE BASIDIOMYCETE COPRINOPSIS
CINEREA 122
A. ABSTRACT 122
B. INTRODUCTION 122
C. MATERIAL AND METHODS 124
IMAGE 3
1. FUNGAL CULTURES 124
2. IMAGING 125
3. ANALYSIS OF THE IMAGES 125
4. DATA PROCESSING 128
D. RESULTS 129
E. DISCUSSION 133
IV. FERMENTATION OF C. CINEREA IN SHAKE FLASKS AND BIOREACTORS 136
A. ABSTRACT 136
B. INTRODUCTION 137
C. MATERIAL AND METHODS 138
1. FUNGAL CULTURES 138
2. DETERMINATION OF THE DRY WEIGHT (DW) IN GRAM PER LITRE [G/L] 139
3. LACCASE ASSAY 140
4. GLUCOSE DETERMINATION 140
5. C/N DETERMINATION OF CULTURE SUPERNATANT 140
6. MORPHOLOGICAL OBSERVATION 140
7. EMBEDDING OF MYCELIUM PELLETS INTO ROTI-PLAST
(FA. ROTH, KARLSRUHE, GERMANY) 141
D. RESULTS 141
1. CULTURAL GROWTH AND LACCASE PRODUCTION AT 25 C AND 37 C 141
2. PELLET MORPHOLOGY IN SHAKE FLASK CULTURES 144
3. MICRO-MORPHOLOGICAL ANALYSIS OF C. CINEREA PELLETS 149
4. LACCASE PRODUCTION IN A STIRRED BIOREACTOR 152
5. PELLET MORPHOLOGY IN STIRRED BIOREACTOR CULTURES 154
E. DISCUSSION 156
1. SHAKE FLASK CULTURES: LACCASE AND BIOMASS PRODUCTION 156
2. SHAKE FLASK CULTURES: PELLET MORPHOLOGY 157
3. FERMENTATION IN A STIRRED BIOREACTOR 161
V. REFERENCES 163
IMAGE 4
CHAPTER 4
NATIVE AND RECOMBINANT LACCASE PRODUCTION WITH COPRINOPSIS CINEREA 171
I. SECRETION OF NATURAL LACCASES IN COPRINOPSIS CINEREA 173
A. ABSTRACT 173
B. INTRODUCTION 173
C. MATERIAL AND METHODS 175
1. C. CINEREA STRAINS AND CULTURE CONDITIONS 175
2. LACCASE ACTIVITY ASSAY 175
3. SDS-PAGE 176
4. STAINING 176
5. PROTEIN IDENTIFICATION 176
D. RESULTS 177
1. LACCASE SECRETION DEPENDS ON MEDIUM AND TEMPERATURE 177 2. DIFFERENT
ISOENZYMES ACCOUNT FOR THE LACCASE ACTIVITY OF DIFFERENT STRAINS 178
E. DISCUSSION 182
1. TEMPERATURE AND MEDIUM EFFECT 182
2. ISOENZYME 183
II. OPTIMISATION OF RECOMBINANT LACCASE PRODUCTION IN COPRINOPSIS
CINEREA 186
A. ABSTRACT 186
B. INTRODUCTION 187
C. MATERIAL AND METHODS 188
1. DNA TRANSFORMATION AND C. CINEREA STRAINS USED 188
2. FUNGAL CULTURES WITH SPORE INOCULUM 188
3. GLUCOSE DETERMINATION 189
4. ERGOSTEROL MEASUREMENT 189
5. DETERMINATION OF THE MYCELIAL DRY WEIGHT (DW) 190
6. ENZYME ASSAYS 190
D. RESULTS 192
1. MEDIA SUPPORTING GROWTH FOR RECOMBINANT LACCASE LED PRODUCTION 192 2.
TEMPERATURE AS AN IMPORTANT GROWTH PARAMETER 198
E. DISCUSSION 203
IMAGE 5
1. MEDIA EFFECTS ON LACCASE PRODUCTION 204
2. TEMPERATURE EFFECTS ON LACCASE SECRETION 208
3. PROTEASE PRODUCTION AND EFFECT ON LACCASE ACTIVTIES 209
III. REFERENCES 210
CHAPTER 5
PURIFICATION AND CHARACTERISATION OF THREE NEW LACCASES FROM THE
BASIDIOMYCETE COPRINOPSIS CINEREA 217
I. ABSTRACT 219
II. INTRODUCTION 220
III. MATERIAL AND METHODS 221
A. FUNGAL CULTURE 221
B. PURIFICATION 221
C. MOLECULAR CHARACTERISATION 223
1. ELECTROPHORESIS 223
2. ISOELECTRIC FOCUSING (IEF) 223
3. STAINING 223
4. PROTEIN IDENTIFICATION 224
D. BIOCHEMICAL CHARACTERISATION 225
1. OPTIMAL REACTION CONDITIONS 226
2. ENZYME KINETICS 226
3. STABILITY OF LACCASES AGAINST INHIBITORS AND IN ORGANIC SOLVENTS 226
4. DYE DECOLOURISATION 227
5. REDOX POTENTIAL DETERMINATION 227
IV. RESULTS 228
A. PURIFICATION 228
B. MOLECULAR CHARACTERISTICS 232
C. BIOCHEMICAL CHARACTERISATION USING STANDARD LACCASE SUBSTRATES 234
D. STABILITY AGAINST INHIBITORS AND ORGANIC SOLVENTS 239
E. REACTIONS WITH INDUSTRIAL DYES 241
F. STRUCTURAL CHARACTERISTICS 242
V
IMAGE 6
V. DISCUSSION 243
A. MOLECULAR STRUCTURE OF ENZYMES PRODUCED BY THE DIFFERENT C. CINEREA
LACCASE GENES 243
B. EFFICIENCY OF THE LACCASE PRODUCTION BY THE DIFFERENT LACCASE
GENES. 246
C. STABILITY OF RECOMBINANTLY PRODUCED ENZYMES 247
D. ENZYMATIC KINETICS WITH DIFFERENT SUBSTRATES 249
E. CONCLUSIONS 252
VI. REFERENCES 253
CHAPTE R6
FINAL DISCUSSION AND PROSPECTS 259
I. SOLID STATE FERMENTATION (SSF) VERSUS SUBMERGED FERMENTATION (SMF)
FOR PRODUCTION OF LACCASES 260
II. IMPROVEMENT OF RECOMBINANT LACCASE PRODUCTION IN COPRINOPSIS CINEREA
262
A. OPTIMISATION OF LACCASE PRODUCTION 263
B. CHANGING BIOCHEMICAL PROPERTIES OF C. CINEREA LACCASES 265
III. GENERAL CONCLUSIONS 266
IV. REFERENCES 267
CURRICULUM VITAE 271
PUBLICATIONS 273 |
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author | Rühl, Martin 1978- |
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dewey-hundreds | 600 - Technology (Applied sciences) 500 - Natural sciences and mathematics |
dewey-ones | 660 - Chemical engineering 572 - Biochemistry |
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discipline | Chemie / Pharmazie Biologie Chemie-Ingenieurwesen Agrar-/Forst-/Ernährungs-/Haushaltswissenschaft / Gartenbau Biotechnologie |
edition | 1. Aufl. |
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spelling | Rühl, Martin 1978- Verfasser (DE-588)140654046 aut Laccases and other ligninolytic enzymes of the basidiomycetes Coprinopsis cinerea and Pleurotus ostreatus submerged and solid state fermentation, morphological studies of liquid cultures and characterisation of new laccases Martin Rühl 1. Aufl. Göttingen Cuvillier Verl. 2010 XII, 275 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Zugl.: Göttingen, Univ., Diss., 2009 Fermentation (DE-588)4129537-7 gnd rswk-swf Coprinus cinereus (DE-588)4391978-9 gnd rswk-swf Laccase (DE-588)4329257-4 gnd rswk-swf Austernseitling (DE-588)4136836-8 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Coprinus cinereus (DE-588)4391978-9 s Laccase (DE-588)4329257-4 s Fermentation (DE-588)4129537-7 s Austernseitling (DE-588)4136836-8 s DE-604 text/html http://deposit.dnb.de/cgi-bin/dokserv?id=3422474&prov=M&dok_var=1&dok_ext=htm Inhaltstext DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=019011542&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
spellingShingle | Rühl, Martin 1978- Laccases and other ligninolytic enzymes of the basidiomycetes Coprinopsis cinerea and Pleurotus ostreatus submerged and solid state fermentation, morphological studies of liquid cultures and characterisation of new laccases Fermentation (DE-588)4129537-7 gnd Coprinus cinereus (DE-588)4391978-9 gnd Laccase (DE-588)4329257-4 gnd Austernseitling (DE-588)4136836-8 gnd |
subject_GND | (DE-588)4129537-7 (DE-588)4391978-9 (DE-588)4329257-4 (DE-588)4136836-8 (DE-588)4113937-9 |
title | Laccases and other ligninolytic enzymes of the basidiomycetes Coprinopsis cinerea and Pleurotus ostreatus submerged and solid state fermentation, morphological studies of liquid cultures and characterisation of new laccases |
title_auth | Laccases and other ligninolytic enzymes of the basidiomycetes Coprinopsis cinerea and Pleurotus ostreatus submerged and solid state fermentation, morphological studies of liquid cultures and characterisation of new laccases |
title_exact_search | Laccases and other ligninolytic enzymes of the basidiomycetes Coprinopsis cinerea and Pleurotus ostreatus submerged and solid state fermentation, morphological studies of liquid cultures and characterisation of new laccases |
title_full | Laccases and other ligninolytic enzymes of the basidiomycetes Coprinopsis cinerea and Pleurotus ostreatus submerged and solid state fermentation, morphological studies of liquid cultures and characterisation of new laccases Martin Rühl |
title_fullStr | Laccases and other ligninolytic enzymes of the basidiomycetes Coprinopsis cinerea and Pleurotus ostreatus submerged and solid state fermentation, morphological studies of liquid cultures and characterisation of new laccases Martin Rühl |
title_full_unstemmed | Laccases and other ligninolytic enzymes of the basidiomycetes Coprinopsis cinerea and Pleurotus ostreatus submerged and solid state fermentation, morphological studies of liquid cultures and characterisation of new laccases Martin Rühl |
title_short | Laccases and other ligninolytic enzymes of the basidiomycetes Coprinopsis cinerea and Pleurotus ostreatus |
title_sort | laccases and other ligninolytic enzymes of the basidiomycetes coprinopsis cinerea and pleurotus ostreatus submerged and solid state fermentation morphological studies of liquid cultures and characterisation of new laccases |
title_sub | submerged and solid state fermentation, morphological studies of liquid cultures and characterisation of new laccases |
topic | Fermentation (DE-588)4129537-7 gnd Coprinus cinereus (DE-588)4391978-9 gnd Laccase (DE-588)4329257-4 gnd Austernseitling (DE-588)4136836-8 gnd |
topic_facet | Fermentation Coprinus cinereus Laccase Austernseitling Hochschulschrift |
url | http://deposit.dnb.de/cgi-bin/dokserv?id=3422474&prov=M&dok_var=1&dok_ext=htm http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=019011542&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
work_keys_str_mv | AT ruhlmartin laccasesandotherligninolyticenzymesofthebasidiomycetescoprinopsiscinereaandpleurotusostreatussubmergedandsolidstatefermentationmorphologicalstudiesofliquidculturesandcharacterisationofnewlaccases |