Optimization of triterpenoid production in engineered Saccharomyces cerevisiae:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Aachen
Apprimus Verlag
2019
|
| Ausgabe: | 1. Auflage |
| Schriftenreihe: | Applied Microbiology
Volume 16 |
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | X, 97 Seiten Illustrationen, Diagramme 21 cm x 14.8 cm, 184 g |
| ISBN: | 9783863597849 3863597842 |
Internformat
MARC
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| 100 | 1 | |a Guo, Hao |d 1988- |e Verfasser |0 (DE-588)1203839162 |4 aut | |
| 245 | 1 | 0 | |a Optimization of triterpenoid production in engineered Saccharomyces cerevisiae |c Hao Guo |
| 250 | |a 1. Auflage | ||
| 264 | 1 | |a Aachen |b Apprimus Verlag |c 2019 | |
| 300 | |a X, 97 Seiten |b Illustrationen, Diagramme |c 21 cm x 14.8 cm, 184 g | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 490 | 1 | |a Applied Microbiology |v Volume 16 | |
| 502 | |b Dissertation |c Rheinisch-Westfälische Technische Hochschule Aachen |d 2019 | ||
| 653 | |a Biologie | ||
| 653 | |a Mikrobiologie | ||
| 653 | |a Triterpenoid | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 710 | 2 | |a Apprimus Verlag |0 (DE-588)1068101474 |4 pbl | |
| 830 | 0 | |a Applied Microbiology |v Volume 16 |w (DE-604)BV042644516 |9 16 | |
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Datensatz im Suchindex
| _version_ | 1804184405440200704 |
|---|---|
| adam_text | CONTENTS
ABSTRACT
..............................................................................................................................................................
V
ZUSAMMENFASSUNG
..........................................................................................................................................
VII
LIST
OF
ABBREVIATIONS
........................................................................................................................................
IX
CHAPTER
1
.............................................................................................................................................................
1
GENERAL
INTRODUCTION
........................................................................................................................................
1
1.1
INTRODUCTION
OF
TRITERPENOIDS
...................................................................................................
3
1.2
S.
CEREVISIAE
A
POTENTIAL
PRODUCTION
HOST
...............................................................................
5
1.3
ENZYMES
OF
THE
TRITERPENOID
PATHWAY
....................................................................................
5
1.3.1
TRITERPENE
SYNTHASES
...................................................................................................................
5
1.3.2
CYTOCHROME
P450
ENZYMES
........................................................................................................6
1.4
METABOLIC
ENGINEERING
OF
TRITERPENOIDS
PRODUCTION
............................................................
9
1.4.1
MEVALONATE
PATHWAY
AND
ERGOSTEROL
PATHWAY
.......................................................................
9
1.4.2
PRECURSOR
SUPPLY
FOR
TRITERPENOID
SYNTHESIS
..........................................................................
11
1.5
STRATEGIES
FOR
MANIPULATING
ERG7
ACTIVITY
.............................................................................
13
1.6
METABOLIC
ENGINEERING
OF
CYP450
.........................................................................................
13
1.7
INFLUENCE
OF
LIPID
DROPLETS
AND
ER
MEMBRANE
ON
TRITERPENOIDS
PRODUCTION
................
15
1.7.1
LIPID
DROPLETS
COMPOSITION
IN
S.
CEREVISIAE
..........................................................................
15
1.7.2
ER
MEMBRANE
AFFECTS
TRITERPENOIDS
PRODUCTION
.....................................................................
15
1.8
SCOPE
OF
THIS
THESIS
...................................................................................................................
16
CHAPTER
2
..........................................................................................................................................................
17
MATERIALS
AND
METHODS
..................................................................................................................................
17
2.1
CHEMICALS
AND
ENZYMES
...........................................................................................................
19
2.2
PLASMIDS
CONSTRUCTION
..............................................................................................................
19
2.2.1
PRIMERS
AND
PLASMIDS
.................................................................................................................
19
2.2
PLASMIDS
CONSTRUCTION
.....................................................................................................
21
2.3
MANIPULATION
OF
ERG7
GENE,
DELETION
OF
AREL
T
ARE2
AND
PAH1
GENE
..........................
23
2.4
YEAST
STRAINS
...............................................................................................................................
23
2.5
MEDIUM
AND
GROWTH
CONDITIONS
............................................................................................
26
2.5.1
MEDIUM
.........................................................................................................................................
26
2.5.2
YEAST
CULTIVATION
CONDITION
.........................................................................................................
27
2.6
BIOREACTOR
PROCESS
...................................................................................................................
27
2.6.1
ETHANOL
RESTRICTION
FEED
PROCESS
...............................................................................................
28
2.7
YEAST
TRANSFORMATION
...............................................................................................................
28
2.8
OPTICAL
DENSITY
MEASUREMENT
................................................................................................
29
TABLE
OF
CONTENTS
2.9
HPLC
ANALYSIS
..............................................................................................................................
29
2.9.1
SAMPLE
PREPARATION
.....................................................................................................................
29
2.9.2
HPLC
CHROMATOGRAPHIC
CONDITION
..............................................................................................
30
2.9.3
GC-MS
ANALYSIS
CONDITION
...........................................................................................................
30
2.9.4
GC-FID
ANALYSIS
CONDITION
...........................................................................................................
30
CHAPTERS
...........................................................................................................................................................
31
INVESTIGATING
THE
IMPACT
OF
THE
ERG7
GENE
OF
BAKER S
YEAST
FOR
HYPER
TRITERPENOID
SYNTHESIS
.........
31
3.1
SUMMARY
.....................................................................................................................................
33
3.2
INTRODUCTION
................................................................................................................................
33
3.3
RESULTS
.........................................................................................................................................
36
3.3.1
MODIFICATION
OF
THE
FRAME-SHIFT
MUTANT
ERG7
IN
SIMOL575
..................................................
36
3.3.2
IMPACT
OF
MUTANT
ERG7
ON
TRITERPENOID
PRODUCTION
AND
STEROL
SYNTHESIS
.........................
37
3.4
DISCUSSION
....................................................................................................................................
38
CHAPTER
4
...........................................................................................................................................................
41
CHASSIS
STRAIN
WITH
MINIMAL
MODIFICATIONS
FOR
HIGH
TRITERPENOID
PRODUCTION
.......................................
41
4.1
SUMMARY
.....................................................................................................................................
43
4.2
INTRODUCTION
................................................................................................................................
43
4.3
RESULTS
.........................................................................................................................................
46
4.3.1
RECONSTRUCTION
OF
2,3-OXIDOSQUALENE
PRODUCER
STRAIN
...........................................................
46
4.3.2
PRODUCTION
OF
LUPANE-TYPE
TRITERPENOIDS
IN
THE
RECONSTRUCTED
2,3-OXIDOSQUALENE
STRAIN48
4.3.3
IMPACT
OF
MUTANT
ERG7
ON
STEROL
SYNTHESIS
.............................................................................
50
4.3.4
INFLUENCE
OF
MEDIUM
COMPOSITION
ON
TRITERPENOIDS
PRODUCTION
.........................................
51
4.3.5
APPLICATION
OF
RECONSTRUCTED
2,3-OXIDOSQUALENE
STRAIN
ON
THE
PRODUCTION
OF
OLEANANE
TYPE
AND
DAMMARANE-TYPE
TRITERPENOIDS
IN
THE
OPTIMIZED
MEDIUM
..............................................
54
4.3.6
EFFECTS
OF
ARE2
AND
ARE1
KNOCKOUTS
ON
THE
PRODUCTION
OF
LUPANE-TYPE
TRITERPENOIDS...
56
4.3.7
PRODUCTION
OF
LUPANE-TYPE,
OLEANANE-TYPE
AND
DAMMARENE-TYPE
TRITERPENOIDS
IN
FED
BATCH
FERMENTATION
.................................................................................................................................
58
4.4
DISCUSSION
...................................................................................................................................
60
CHAPTERS
...........................................................................................................................................................
63
ATTEMPTING
TO
OPTIMIZE
CYTOCHROME
P450
OXIDATION
................................................................................
63
5.1
SUMMARY
...................................................................................................................................
64
5.2
INTRODUCTION
...............................................................................................................................
65
5.3
RESULTS
........................................................................................................................................
66
5.4
DISCUSSION
..................................................................................................................................
70
CHAPTER
6
...........................................................................................................................................................
73
GENERAL
DISCUSSION
...........................................................................................................................................
73
II
TABLE
OF
CONTENTS
CHAPTER
7
..........................................................................................................................................................
81
CONCLUSION
AND
OUTLOOK
..................................................................................................................................
81
7.1
CONCLUSION
..................................................................................................................................
83
7.2
OUTLOOK
........................................................................................................................................
83
REFERENCE
.........................................................................................................................................................
85
III
|
| adam_txt |
CONTENTS
ABSTRACT
.
V
ZUSAMMENFASSUNG
.
VII
LIST
OF
ABBREVIATIONS
.
IX
CHAPTER
1
.
1
GENERAL
INTRODUCTION
.
1
1.1
INTRODUCTION
OF
TRITERPENOIDS
.
3
1.2
S.
CEREVISIAE
A
POTENTIAL
PRODUCTION
HOST
.
5
1.3
ENZYMES
OF
THE
TRITERPENOID
PATHWAY
.
5
1.3.1
TRITERPENE
SYNTHASES
.
5
1.3.2
CYTOCHROME
P450
ENZYMES
.6
1.4
METABOLIC
ENGINEERING
OF
TRITERPENOIDS
PRODUCTION
.
9
1.4.1
MEVALONATE
PATHWAY
AND
ERGOSTEROL
PATHWAY
.
9
1.4.2
PRECURSOR
SUPPLY
FOR
TRITERPENOID
SYNTHESIS
.
11
1.5
STRATEGIES
FOR
MANIPULATING
ERG7
ACTIVITY
.
13
1.6
METABOLIC
ENGINEERING
OF
CYP450
.
13
1.7
INFLUENCE
OF
LIPID
DROPLETS
AND
ER
MEMBRANE
ON
TRITERPENOIDS
PRODUCTION
.
15
1.7.1
LIPID
DROPLETS
COMPOSITION
IN
S.
CEREVISIAE
.
15
1.7.2
ER
MEMBRANE
AFFECTS
TRITERPENOIDS
PRODUCTION
.
15
1.8
SCOPE
OF
THIS
THESIS
.
16
CHAPTER
2
.
17
MATERIALS
AND
METHODS
.
17
2.1
CHEMICALS
AND
ENZYMES
.
19
2.2
PLASMIDS
CONSTRUCTION
.
19
2.2.1
PRIMERS
AND
PLASMIDS
.
19
2.2
PLASMIDS
CONSTRUCTION
.
21
2.3
MANIPULATION
OF
ERG7
GENE,
DELETION
OF
AREL
T
ARE2
AND
PAH1
GENE
.
23
2.4
YEAST
STRAINS
.
23
2.5
MEDIUM
AND
GROWTH
CONDITIONS
.
26
2.5.1
MEDIUM
.
26
2.5.2
YEAST
CULTIVATION
CONDITION
.
27
2.6
BIOREACTOR
PROCESS
.
27
2.6.1
ETHANOL
RESTRICTION
FEED
PROCESS
.
28
2.7
YEAST
TRANSFORMATION
.
28
2.8
OPTICAL
DENSITY
MEASUREMENT
.
29
TABLE
OF
CONTENTS
2.9
HPLC
ANALYSIS
.
29
2.9.1
SAMPLE
PREPARATION
.
29
2.9.2
HPLC
CHROMATOGRAPHIC
CONDITION
.
30
2.9.3
GC-MS
ANALYSIS
CONDITION
.
30
2.9.4
GC-FID
ANALYSIS
CONDITION
.
30
CHAPTERS
.
31
INVESTIGATING
THE
IMPACT
OF
THE
ERG7
GENE
OF
BAKER'S
YEAST
FOR
HYPER
TRITERPENOID
SYNTHESIS
.
31
3.1
SUMMARY
.
33
3.2
INTRODUCTION
.
33
3.3
RESULTS
.
36
3.3.1
MODIFICATION
OF
THE
FRAME-SHIFT
MUTANT
ERG7
IN
SIMOL575
.
36
3.3.2
IMPACT
OF
MUTANT
ERG7
ON
TRITERPENOID
PRODUCTION
AND
STEROL
SYNTHESIS
.
37
3.4
DISCUSSION
.
38
CHAPTER
4
.
41
CHASSIS
STRAIN
WITH
MINIMAL
MODIFICATIONS
FOR
HIGH
TRITERPENOID
PRODUCTION
.
41
4.1
SUMMARY
.
43
4.2
INTRODUCTION
.
43
4.3
RESULTS
.
46
4.3.1
RECONSTRUCTION
OF
2,3-OXIDOSQUALENE
PRODUCER
STRAIN
.
46
4.3.2
PRODUCTION
OF
LUPANE-TYPE
TRITERPENOIDS
IN
THE
RECONSTRUCTED
2,3-OXIDOSQUALENE
STRAIN48
4.3.3
IMPACT
OF
MUTANT
ERG7
ON
STEROL
SYNTHESIS
.
50
4.3.4
INFLUENCE
OF
MEDIUM
COMPOSITION
ON
TRITERPENOIDS
PRODUCTION
.
51
4.3.5
APPLICATION
OF
RECONSTRUCTED
2,3-OXIDOSQUALENE
STRAIN
ON
THE
PRODUCTION
OF
OLEANANE
TYPE
AND
DAMMARANE-TYPE
TRITERPENOIDS
IN
THE
OPTIMIZED
MEDIUM
.
54
4.3.6
EFFECTS
OF
ARE2
AND
ARE1
KNOCKOUTS
ON
THE
PRODUCTION
OF
LUPANE-TYPE
TRITERPENOIDS.
56
4.3.7
PRODUCTION
OF
LUPANE-TYPE,
OLEANANE-TYPE
AND
DAMMARENE-TYPE
TRITERPENOIDS
IN
FED
BATCH
FERMENTATION
.
58
4.4
DISCUSSION
.
60
CHAPTERS
.
63
ATTEMPTING
TO
OPTIMIZE
CYTOCHROME
P450
OXIDATION
.
63
5.1
SUMMARY
.
64
5.2
INTRODUCTION
.
65
5.3
RESULTS
.
66
5.4
DISCUSSION
.
70
CHAPTER
6
.
73
GENERAL
DISCUSSION
.
73
II
TABLE
OF
CONTENTS
CHAPTER
7
.
81
CONCLUSION
AND
OUTLOOK
.
81
7.1
CONCLUSION
.
83
7.2
OUTLOOK
.
83
REFERENCE
.
85
III |
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| author | Guo, Hao 1988- |
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| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV048462682 |
| illustrated | Illustrated |
| index_date | 2024-07-03T20:34:06Z |
| indexdate | 2024-07-10T09:38:49Z |
| institution | BVB |
| institution_GND | (DE-588)1068101474 |
| isbn | 9783863597849 3863597842 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-033840651 |
| oclc_num | 1196189553 |
| open_access_boolean | |
| owner | DE-29T |
| owner_facet | DE-29T |
| physical | X, 97 Seiten Illustrationen, Diagramme 21 cm x 14.8 cm, 184 g |
| publishDate | 2019 |
| publishDateSearch | 2019 |
| publishDateSort | 2019 |
| publisher | Apprimus Verlag |
| record_format | marc |
| series | Applied Microbiology |
| series2 | Applied Microbiology |
| spelling | Guo, Hao 1988- Verfasser (DE-588)1203839162 aut Optimization of triterpenoid production in engineered Saccharomyces cerevisiae Hao Guo 1. Auflage Aachen Apprimus Verlag 2019 X, 97 Seiten Illustrationen, Diagramme 21 cm x 14.8 cm, 184 g txt rdacontent n rdamedia nc rdacarrier Applied Microbiology Volume 16 Dissertation Rheinisch-Westfälische Technische Hochschule Aachen 2019 Biologie Mikrobiologie Triterpenoid (DE-588)4113937-9 Hochschulschrift gnd-content Apprimus Verlag (DE-588)1068101474 pbl Applied Microbiology Volume 16 (DE-604)BV042644516 16 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=033840651&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p vlb 20220810 DE-101 https://d-nb.info/provenance/plan#vlb |
| spellingShingle | Guo, Hao 1988- Optimization of triterpenoid production in engineered Saccharomyces cerevisiae Applied Microbiology |
| subject_GND | (DE-588)4113937-9 |
| title | Optimization of triterpenoid production in engineered Saccharomyces cerevisiae |
| title_auth | Optimization of triterpenoid production in engineered Saccharomyces cerevisiae |
| title_exact_search | Optimization of triterpenoid production in engineered Saccharomyces cerevisiae |
| title_exact_search_txtP | Optimization of triterpenoid production in engineered Saccharomyces cerevisiae |
| title_full | Optimization of triterpenoid production in engineered Saccharomyces cerevisiae Hao Guo |
| title_fullStr | Optimization of triterpenoid production in engineered Saccharomyces cerevisiae Hao Guo |
| title_full_unstemmed | Optimization of triterpenoid production in engineered Saccharomyces cerevisiae Hao Guo |
| title_short | Optimization of triterpenoid production in engineered Saccharomyces cerevisiae |
| title_sort | optimization of triterpenoid production in engineered saccharomyces cerevisiae |
| topic_facet | Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=033840651&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| volume_link | (DE-604)BV042644516 |
| work_keys_str_mv | AT guohao optimizationoftriterpenoidproductioninengineeredsaccharomycescerevisiae AT apprimusverlag optimizationoftriterpenoidproductioninengineeredsaccharomycescerevisiae |