Metabolic engineering of pseudomonas taiwanensis VLB120 for sustainable production of 4-Hydroxybenzoate:
Saved in:
| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
Aachen
Apprimus Verlag
2020
|
| Edition: | 1. Auflage |
| Series: | Applied microbiology
Volume 17 |
| Subjects: | |
| Online Access: | Volltext Inhaltstext Inhaltsverzeichnis Inhaltsverzeichnis |
| Physical Description: | XVIII, 153 Seiten Illustrationen, Diagramme 21 cm, 254 g |
| ISBN: | 9783863598167 3863598164 |
| DOI: | 10.18154/RWTH-2020-02978 |
Staff View
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| 245 | 1 | 0 | |a Metabolic engineering of pseudomonas taiwanensis VLB120 for sustainable production of 4-Hydroxybenzoate |c Christoph Lenzen |
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| adam_text | TABLE OF CONTENTS
TABLE OF CONTENTS
LIST OF FIGURES.V
LIST OF TABLES.VII
LIST OF ABBREVIATIONS.IX
SUMMARY .XV
ZUSAMMENFASSUNG.XVII
1 GENERAL INTRODUCTION.3
1.1 BIOTECHNOLOGY - A SCIENCE APPLIED IN EVERYDAY LIFE.3
1.2 BIOCATALYSIS AS AN ALTERNATIVE TO CHEMICAL SYNTHESIS.5
1.3 METABOLIC ENGINEERING - RATIONAL VS. NON-RATIONAL STRATEGIES. 6
1.4 4-HYDROXYBENZOATE - A COMPOUND OF INDUSTRIAL RELEVANCE.8
1.5 PSEUDOMONAS AS A MICROBIAL CELL FACTORY.12
1.6 SCOPE AND OUTLINE OF THIS THESIS.14
2 MATERIALS AND METHODS.19
2.1 CHEMICALS.19
2.2 BACTERIAL STRAINS AND CULTURE CONDITIONS.19
2.2.1 PULSED FED-BATCH CULTIVATION.22
2.2.2 CARBON-LIMITED CHEMOSTAT CULTIVATION.22
2.3 DNA TECHNIQUES.23
2.4 MNNG MUTAGENESIS.25
2.5 ANALYTICAL METHODS.31
2.5.1 AGAROSE GEL ELECTROPHORESIS.31
2.5.2 HPLC ANALYSIS.32
2.5.3 FLUORESCENCE DETECTION AND HIGH-THROUGHPUT SCREENING.32
2.5.4 METABOLIC MODELING.33
3 RESULTS.37
3.1 RATIONAL METABOLIC ENGINEERING OF P. TAIWANENSIS VLB120 FOR THE
BIOCONVERSION OF 4-
HYDROXYBENZOATE.37
3.1.1 SUMMARY.38
3.1.2 INTRODUCTION.39
3.1.3 RESULTS AND DISCUSSION.-.41
3.1.3.1 ESTABLISHING 4-HYDROXYBENZOATE PRODUCTION IN P. TAIWANENSIS.41
3.1.3.2 ENHANCING SUPPLY OF TYROSINE.44
I
BIBLIOGRAFISCHE INFORMATIONEN
HTTP://D-NB.INFO/1204719128
TABLE OF CONTENTS
3.1.3.3 STABLE GENOMIC INTEGRATION OF PRODUCTION MODULES.46
3.1.3.4 INVESTIGATION OF THE EFFECT OF CATABOLITE REPRESSION ON
4-HYDROXYBENZOATE
PRODUCTION.47
3.1.3.5 DEREPRESSION OF GLUCOSE CONSUMPTION AND TYROSINE BIOSYNTHESIS -
KNOCKOUT OF
HEXR AND TYRR.50
3.1.3.6 ENHANCED 4-HYDROXYBENZOATE PRODUCTION THROUGH OVEREXPRESSION OF
PPSA
AND PGI.52
3.1.3.7 PRODUCTION OF 4-HYDROXYBENZOATE IS ENHANCED ON GLYCEROL.54
3.1.3.8 PRODUCTION OF 4-HYDROXYBENZOATE FROM GLYCEROL VIA PULSED
FED-BATCH CULTIVATION.57
3.1.4 CONCLUSION.60
3.2 DEVELOPMENT OF A FLUORESCENCE-BASED BIOSENSOR FOR HIGH-THROUGHPUT
SCREENING OF A
MUTANT LIBRARY OF P. TAIWANENSIS VLB120 FOR ENHANCED BIOCONVERSION OF 4-
HYDROXYBENZOATE.61
3.2.1 SUMMARY.62
3.2.2 INTRODUCTION.63
3.2.3 RESULTS AND DISCUSSION. .66
3.2.3.1 BIOSENSOR ENGINEERING.66
3.2.3.2 MUTAGENESIS AND HIGH-THROUGHPUT SCREENING VIA FACS.71
3.2.3.3 BIOSENSOR OPTIMIZATION.77
3.2.4 CONCLUSION.81
3.3 ESTABLISHMENT OF METABOLIC DEMAND FOR ACETYL-COA IN P. TAIWANENSIS
VLB120 FOR
ENHANCED PRODUCTION OF 4-HYDROXYBENZOATE.83
3.3.1 SUMMARY...84
3.3.2 INTRODUCTION.85
3.3.3 RESULTS AND DISCUSSION.87
3.3.3.1 IN SILLICO EVALUATION OF METABOLIC DEMAND VIA 4-HYDROXYBENZOATE
PRODUCTION ...89
3.3.3.2 IN VIVO EVALUATION OF METABOLIC DEMAND VIA 4-HYDROXYBENZOATE
PRODUCTION.91
3.3.4 CONCLUSION.96
4 GENERAL DISCUSSION AND OUTLOOK.99
4.1 RATIONAL BIOCATALYST DESIGN.100
4.2 NON-RATIONAL ENGINEERING - BIOSENSOR DEVELOPMENT.105
4.3 NON-RATIONAL ENGINEERING - METABOLIC DEMAND.107
4.4 CONCLUSION.108
5 REFERENCES.ILL
6 APPENDIX.145
6.1 CODON-OPTIMIZED GENES.145
6.1.1 TYROSINE AMMONIA-LYASE [RHODOBACTER SPHAEROIDES).145
II
TABLE OF CONTENTS
6.1.2 TYROSINE AMMONIA-LYASE (FLAVOBACTERIUM JOHNSONIAE).146
6.1.3 PHENYLALANINE AMMONIA-LYASE (TRICHOSPORON CUTANEUM).147
7 CURRICULUM VITAE.153
III
|
| adam_txt |
TABLE OF CONTENTS
TABLE OF CONTENTS
LIST OF FIGURES.V
LIST OF TABLES.VII
LIST OF ABBREVIATIONS.IX
SUMMARY .XV
ZUSAMMENFASSUNG.XVII
1 GENERAL INTRODUCTION.3
1.1 BIOTECHNOLOGY - A SCIENCE APPLIED IN EVERYDAY LIFE.3
1.2 BIOCATALYSIS AS AN ALTERNATIVE TO CHEMICAL SYNTHESIS.5
1.3 METABOLIC ENGINEERING - RATIONAL VS. NON-RATIONAL STRATEGIES. 6
1.4 4-HYDROXYBENZOATE - A COMPOUND OF INDUSTRIAL RELEVANCE.8
1.5 PSEUDOMONAS AS A MICROBIAL CELL FACTORY.12
1.6 SCOPE AND OUTLINE OF THIS THESIS.14
2 MATERIALS AND METHODS.19
2.1 CHEMICALS.19
2.2 BACTERIAL STRAINS AND CULTURE CONDITIONS.19
2.2.1 PULSED FED-BATCH CULTIVATION.22
2.2.2 CARBON-LIMITED CHEMOSTAT CULTIVATION.22
2.3 DNA TECHNIQUES.23
2.4 MNNG MUTAGENESIS.25
2.5 ANALYTICAL METHODS.31
2.5.1 AGAROSE GEL ELECTROPHORESIS.31
2.5.2 HPLC ANALYSIS.32
2.5.3 FLUORESCENCE DETECTION AND HIGH-THROUGHPUT SCREENING.32
2.5.4 METABOLIC MODELING.33
3 RESULTS.37
3.1 RATIONAL METABOLIC ENGINEERING OF P. TAIWANENSIS VLB120 FOR THE
BIOCONVERSION OF 4-
HYDROXYBENZOATE.37
3.1.1 SUMMARY.38
3.1.2 INTRODUCTION.39
3.1.3 RESULTS AND DISCUSSION.-.41
3.1.3.1 ESTABLISHING 4-HYDROXYBENZOATE PRODUCTION IN P. TAIWANENSIS.41
3.1.3.2 ENHANCING SUPPLY OF TYROSINE.44
I
BIBLIOGRAFISCHE INFORMATIONEN
HTTP://D-NB.INFO/1204719128
TABLE OF CONTENTS
3.1.3.3 STABLE GENOMIC INTEGRATION OF PRODUCTION MODULES.46
3.1.3.4 INVESTIGATION OF THE EFFECT OF CATABOLITE REPRESSION ON
4-HYDROXYBENZOATE
PRODUCTION.47
3.1.3.5 DEREPRESSION OF GLUCOSE CONSUMPTION AND TYROSINE BIOSYNTHESIS -
KNOCKOUT OF
HEXR AND TYRR.50
3.1.3.6 ENHANCED 4-HYDROXYBENZOATE PRODUCTION THROUGH OVEREXPRESSION OF
PPSA
AND PGI.52
3.1.3.7 PRODUCTION OF 4-HYDROXYBENZOATE IS ENHANCED ON GLYCEROL.54
3.1.3.8 PRODUCTION OF 4-HYDROXYBENZOATE FROM GLYCEROL VIA PULSED
FED-BATCH CULTIVATION.57
3.1.4 CONCLUSION.60
3.2 DEVELOPMENT OF A FLUORESCENCE-BASED BIOSENSOR FOR HIGH-THROUGHPUT
SCREENING OF A
MUTANT LIBRARY OF P. TAIWANENSIS VLB120 FOR ENHANCED BIOCONVERSION OF 4-
HYDROXYBENZOATE.61
3.2.1 SUMMARY.62
3.2.2 INTRODUCTION.63
3.2.3 RESULTS AND DISCUSSION. .66
3.2.3.1 BIOSENSOR ENGINEERING.66
3.2.3.2 MUTAGENESIS AND HIGH-THROUGHPUT SCREENING VIA FACS.71
3.2.3.3 BIOSENSOR OPTIMIZATION.77
3.2.4 CONCLUSION.81
3.3 ESTABLISHMENT OF METABOLIC DEMAND FOR ACETYL-COA IN P. TAIWANENSIS
VLB120 FOR
ENHANCED PRODUCTION OF 4-HYDROXYBENZOATE.83
3.3.1 SUMMARY.84
3.3.2 INTRODUCTION.85
3.3.3 RESULTS AND DISCUSSION.87
3.3.3.1 IN SILLICO EVALUATION OF METABOLIC DEMAND VIA 4-HYDROXYBENZOATE
PRODUCTION .89
3.3.3.2 IN VIVO EVALUATION OF METABOLIC DEMAND VIA 4-HYDROXYBENZOATE
PRODUCTION.91
3.3.4 CONCLUSION.96
4 GENERAL DISCUSSION AND OUTLOOK.99
4.1 RATIONAL BIOCATALYST DESIGN.100
4.2 NON-RATIONAL ENGINEERING - BIOSENSOR DEVELOPMENT.105
4.3 NON-RATIONAL ENGINEERING - METABOLIC DEMAND.107
4.4 CONCLUSION.108
5 REFERENCES.ILL
6 APPENDIX.145
6.1 CODON-OPTIMIZED GENES.145
6.1.1 TYROSINE AMMONIA-LYASE [RHODOBACTER SPHAEROIDES).145
II
TABLE OF CONTENTS
6.1.2 TYROSINE AMMONIA-LYASE (FLAVOBACTERIUM JOHNSONIAE).146
6.1.3 PHENYLALANINE AMMONIA-LYASE (TRICHOSPORON CUTANEUM).147
7 CURRICULUM VITAE.153
III |
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| dewey-full | 660.63 |
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| dewey-sort | 3660.63 |
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| discipline | Chemie / Pharmazie |
| discipline_str_mv | Chemie / Pharmazie |
| doi_str_mv | 10.18154/RWTH-2020-02978 |
| edition | 1. Auflage |
| format | Thesis Book |
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| oclc_num | 1164615444 |
| open_access_boolean | 1 |
| owner | DE-29T |
| owner_facet | DE-29T |
| physical | XVIII, 153 Seiten Illustrationen, Diagramme 21 cm, 254 g |
| psigel | ebook |
| publishDate | 2020 |
| publishDateSearch | 2020 |
| publishDateSort | 2020 |
| publisher | Apprimus Verlag |
| record_format | marc |
| series | Applied microbiology |
| series2 | Applied microbiology |
| spelling | Lenzen, Christoph 1984- Verfasser (DE-588)1205940316 aut Metabolic engineering of pseudomonas taiwanensis VLB120 for sustainable production of 4-Hydroxybenzoate Christoph Lenzen 1. Auflage Aachen Apprimus Verlag 2020 XVIII, 153 Seiten Illustrationen, Diagramme 21 cm, 254 g txt rdacontent n rdamedia nc rdacarrier Applied microbiology Volume 17 Dissertation RWTH Aachen University 2019 Pseudomonas taiwanensis (DE-588)1064240615 gnd rswk-swf Hydroxybenzoesäureester 4- (DE-588)4286645-5 gnd rswk-swf Herstellung (DE-588)4159653-5 gnd rswk-swf Biokonversion (DE-588)4145610-5 gnd rswk-swf Naturwissenschaften Metabolic Engineering (DE-588)4113937-9 Hochschulschrift gnd-content Hydroxybenzoesäureester 4- (DE-588)4286645-5 s Herstellung (DE-588)4159653-5 s Biokonversion (DE-588)4145610-5 s Pseudomonas taiwanensis (DE-588)1064240615 s DE-604 Apprimus Verlag (DE-588)1068101474 pbl Erscheint auch als Online-Ausgabe http://doi.org/10.18154/RWTH-2020-02978 Applied microbiology Volume 17 (DE-604)BV042644516 17 http://doi.org/10.18154/RWTH-2020-02978 Resolving-System kostenfrei Volltext X:MVB text/html http://deposit.dnb.de/cgi-bin/dokserv?id=9fb2e9fc5b2446b99fd58c960aa6599b&prov=M&dok_var=1&dok_ext=htm Inhaltstext B:DE-101 application/pdf https://d-nb.info/1204719128/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032187517&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Lenzen, Christoph 1984- Metabolic engineering of pseudomonas taiwanensis VLB120 for sustainable production of 4-Hydroxybenzoate Applied microbiology Pseudomonas taiwanensis (DE-588)1064240615 gnd Hydroxybenzoesäureester 4- (DE-588)4286645-5 gnd Herstellung (DE-588)4159653-5 gnd Biokonversion (DE-588)4145610-5 gnd |
| subject_GND | (DE-588)1064240615 (DE-588)4286645-5 (DE-588)4159653-5 (DE-588)4145610-5 (DE-588)4113937-9 |
| title | Metabolic engineering of pseudomonas taiwanensis VLB120 for sustainable production of 4-Hydroxybenzoate |
| title_auth | Metabolic engineering of pseudomonas taiwanensis VLB120 for sustainable production of 4-Hydroxybenzoate |
| title_exact_search | Metabolic engineering of pseudomonas taiwanensis VLB120 for sustainable production of 4-Hydroxybenzoate |
| title_exact_search_txtP | Metabolic engineering of pseudomonas taiwanensis VLB120 for sustainable production of 4-Hydroxybenzoate |
| title_full | Metabolic engineering of pseudomonas taiwanensis VLB120 for sustainable production of 4-Hydroxybenzoate Christoph Lenzen |
| title_fullStr | Metabolic engineering of pseudomonas taiwanensis VLB120 for sustainable production of 4-Hydroxybenzoate Christoph Lenzen |
| title_full_unstemmed | Metabolic engineering of pseudomonas taiwanensis VLB120 for sustainable production of 4-Hydroxybenzoate Christoph Lenzen |
| title_short | Metabolic engineering of pseudomonas taiwanensis VLB120 for sustainable production of 4-Hydroxybenzoate |
| title_sort | metabolic engineering of pseudomonas taiwanensis vlb120 for sustainable production of 4 hydroxybenzoate |
| topic | Pseudomonas taiwanensis (DE-588)1064240615 gnd Hydroxybenzoesäureester 4- (DE-588)4286645-5 gnd Herstellung (DE-588)4159653-5 gnd Biokonversion (DE-588)4145610-5 gnd |
| topic_facet | Pseudomonas taiwanensis Hydroxybenzoesäureester 4- Herstellung Biokonversion Hochschulschrift |
| url | http://doi.org/10.18154/RWTH-2020-02978 http://deposit.dnb.de/cgi-bin/dokserv?id=9fb2e9fc5b2446b99fd58c960aa6599b&prov=M&dok_var=1&dok_ext=htm https://d-nb.info/1204719128/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032187517&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| volume_link | (DE-604)BV042644516 |
| work_keys_str_mv | AT lenzenchristoph metabolicengineeringofpseudomonastaiwanensisvlb120forsustainableproductionof4hydroxybenzoate AT apprimusverlag metabolicengineeringofpseudomonastaiwanensisvlb120forsustainableproductionof4hydroxybenzoate |