Engineering formatotrophic growth in Escherichia coli:
Saved in:
| Main Author: | |
|---|---|
| Format: | Thesis Book |
| Language: | English |
| Published: |
Potsdam
[2020]
|
| Subjects: | |
| Online Access: | Inhaltsverzeichnis Inhaltsverzeichnis |
| Physical Description: | V, 107 Seiten Illustrationen 30 cm |
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MARC
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| 100 | 1 | |a Wenk, Sebastian |e Verfasser |0 (DE-588)1220117633 |4 aut | |
| 245 | 1 | 0 | |a Engineering formatotrophic growth in Escherichia coli |c Sebastian Wenk |
| 264 | 1 | |a Potsdam |c [2020] | |
| 300 | |a V, 107 Seiten |b Illustrationen |c 30 cm | ||
| 336 | |b txt |2 rdacontent | ||
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| 502 | |b Dissertation |c Universität Potsdam |d 2020 | ||
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Record in the Search Index
| _version_ | 1804182547345702912 |
|---|---|
| adam_text | TABLE
OF
CONTENT
OVERARCHING
INTRODUCTION
..........................................................................................................
1
CO
2
-GREENHOUSE
GAS
AND
VALUABLE
FEEDSTOCK
...............................................................................
1
BIOLOGICAL
CO2
AND
CI
FIXATION
.........................................................................................................
2
CO2
CAPTURE
AND
CONVERSION
INTO
FORMATE
......................................................................................
4
TOWARDS
A
SUSTAINABLE
BIOECONOMY
-
SYNTHETIC
CI
ASSIMILATION
...................................................
6
SYNTHETIC
FORMATE
ASSIMILATION
PATHWAYS
.......................................................................................
7
AIM
OF
THIS
WORK
...............................................................................................................................
9
MATERIALS
.............................................................................................................................................
11
REAGENTS
........................................................................................................................................
11
GROWTH
MEDIA
................................................................................................................................
11
BACTERIAL
STRAINS
.............................................................................................................................
11
METHODS
...............................................................................................................................................
16
GENOME
ENGINEERING
.....................................................................................................................
16
SYNTHETIC
OPERON
CONSTRUCTION
.....................................................................................................
17
GROWTH
CHARACTERIZATION
IN
MICROPLATE
READERS
...........................................................................
18
ANALYSIS
OF
SUPERNATANT
BY
ION
CHROMATOGRAPHY
..........................................................................
18
13
C-LABELLING
OF
PROTEINOGENIC
AMINO
ACIDS
...................................................................................
19
DRY
WEIGHT
ANALYSIS
.................................................................
19
ENZYMES
AND
CHEMICAL
ASSAYS
.......................................................................................................
20
QUANTITATIVE
POLYMERASE
CHAIN
REACTION
......................................................................................
20
CHAPTER
1:
ESTABLISHING
A
STRATEGY
AND
METHODS
FOR
METABOLIC
ENGINEERING
OF
E.
COU
........
.
..............................
............
...21
STRAIN
ENGINEERING:
IN
VIVO
SELECTION
FOR
PATHWAY
MODULES
.........................................................
23
PATHWAY
ENGINEERING:
FINE-TUNE
EXPRESSION
OF
ENZYMES
.............................................................
25
GENOMIC
INTEGRATION
OF
SYNTHETIC
OPERONS
...................................................................................
27
GROWTH
EXPERIMENTS:
IN
VIVO
SELECTION
FOR
PATHWAY
ACTIVITY
.......................................................
29
PATHWAY
CONFIRMATION:
13
C-LABELING
OF
PROTEINOGENIC
AMINO
ACIDS
.............................................
32
CHAPTER
2:
AN
ENERGY
AUXOTROPH
E.
COU
TO
TEST
THE
IN
VIVO
ACTIVITY
OF
CI-
OXIDIZING
ENZYMES......
............................................
33
CONSTRUCTION
AND
VALIDATION
OF
AN
E.
COLI
STRAIN
AUXOTROPH
TO
REDUCING
POWER
AND
ENERGY
......
34
TESTING
FORMATE
AS
A
REDUCING
POWER
SOURCE
...............................................................................
37
TESTING
METHANOL
AS
A
REDUCING
POWER
SOURCE
.............................................................................
38
CHAPTER
3:
ESTABLISHING
THE
SERINE
THREONINE
CYCLE
IN
COLL
...................
.............
43
PRODUCTION
OF
ACETYL-COA,
GLYCINE
AND
SERINE
VIA
THREONINE
SYNTHESIS
AND
CLEAVAGE
...............
45
ESTABLISHMENT
OF
AN
EXTERNAL
ENERGY
PRODUCTION
MODULE
IMPROVES
GROWTH
ON
GLYCEROL
AND
ALLOWS
GROWTH
ON
PYRUVATE
............................................................................................................
49
FORMATE
IS
ASSIMILATED
TO
ALLOW
GROWTH
VIA
THE
SERINE
THREONINE
CYCLE
......................................
51
SHORT-TERM
EVOLUTION
IMPROVES
GROWTH
ON
PYRUVATE
AND
FORMATE
AND
ENABLES
GROWTH
ON
GLYCINE
AND
FORMATE
....................................................................................................................
54
CHAPTER
4:
GROWTH
OF
E.
COU
ON
FORMATE
AND
METHANOL
VIA
THE
REDUCTIVE
GLYCINE
PATHWAY
......................
57
THE
REDUCTIVE
GLYCINE
PATHWAY
......................................................................................................
59
MODULAR
ENGINEERING
ESTABLISHES
GROWTH
ON
FORMATE
..................................................................
61
SHORT-TERM
EVOLUTION
IMPROVES
GROWTH
ON
FORMATE
....................................................................
64
CARBON
LABELLING
SHEDS
LIGHT
ON
CELLULAR
FLUXES
.............................................................................
66
ENGINEERED
GROWTH
OF
E.
COLI
ON
METHANOL
...................................................................................
67
DISCUSSION
............................................................................................................................................
71
SYNTHETIC
BACTERIAL
METABOLISM
.....................................................................................................
71
THE
POWER
OF
ADAPTIVE
LABORATORY
EVOLUTION
...............................................................................
72
MEETING
CELLULAR
ENERGY
DEMANDS
BY
CI
OXIDATION
.......................................................................
73
FORMATOTROPHIC
GROWTH
VIA
THE
REDUCTIVE
GLYCINE
PATHWAY
.........................................................
74
FORMATOTROPHIC
GROWTH
VIA
THE
SERINE
THREONINE
CYCLE
...............................................................
75
OUTLOOK
-
THE
FUTURE
OF
SYNTHETIC
CI
METABOLISM
........................................................................
77
APPENDIX
...............................................................................................................................................
79
SUPPLEMENTARY
FIGURES
..................................................................................................................
79
REFERENCES
............................................................................................................................................
89
DISCLAIMER
..........................................................................................................................................
103
DECLARATION
OF
WORK
...................................................................................................................
105
EIDESSTATTLICHE
ERKLARUNG
........................................................................................................
105
ACKNOWLEDGEMENTS
.....................................................................................................................
107
II
|
| adam_txt |
TABLE
OF
CONTENT
OVERARCHING
INTRODUCTION
.
1
CO
2
-GREENHOUSE
GAS
AND
VALUABLE
FEEDSTOCK
.
1
BIOLOGICAL
CO2
AND
CI
FIXATION
.
2
CO2
CAPTURE
AND
CONVERSION
INTO
FORMATE
.
4
TOWARDS
A
SUSTAINABLE
BIOECONOMY
-
SYNTHETIC
CI
ASSIMILATION
.
6
SYNTHETIC
FORMATE
ASSIMILATION
PATHWAYS
.
7
AIM
OF
THIS
WORK
.
9
MATERIALS
.
11
REAGENTS
.
11
GROWTH
MEDIA
.
11
BACTERIAL
STRAINS
.
11
METHODS
.
16
GENOME
ENGINEERING
.
16
SYNTHETIC
OPERON
CONSTRUCTION
.
17
GROWTH
CHARACTERIZATION
IN
MICROPLATE
READERS
.
18
ANALYSIS
OF
SUPERNATANT
BY
ION
CHROMATOGRAPHY
.
18
13
C-LABELLING
OF
PROTEINOGENIC
AMINO
ACIDS
.
19
DRY
WEIGHT
ANALYSIS
.
19
ENZYMES
AND
CHEMICAL
ASSAYS
.
20
QUANTITATIVE
POLYMERASE
CHAIN
REACTION
.
20
CHAPTER
1:
ESTABLISHING
A
STRATEGY
AND
METHODS
FOR
METABOLIC
ENGINEERING
OF
E.
COU
.
.
.
.
.21
STRAIN
ENGINEERING:
IN
VIVO
SELECTION
FOR
PATHWAY
MODULES
.
23
PATHWAY
ENGINEERING:
FINE-TUNE
EXPRESSION
OF
ENZYMES
.
25
GENOMIC
INTEGRATION
OF
SYNTHETIC
OPERONS
.
27
GROWTH
EXPERIMENTS:
IN
VIVO
SELECTION
FOR
PATHWAY
ACTIVITY
.
29
PATHWAY
CONFIRMATION:
13
C-LABELING
OF
PROTEINOGENIC
AMINO
ACIDS
.
32
CHAPTER
2:
AN
ENERGY
AUXOTROPH
E.
COU
TO
TEST
THE
IN
VIVO
ACTIVITY
OF
CI-
OXIDIZING
ENZYMES.
.
33
CONSTRUCTION
AND
VALIDATION
OF
AN
E.
COLI
STRAIN
AUXOTROPH
TO
REDUCING
POWER
AND
ENERGY
.
34
TESTING
FORMATE
AS
A
REDUCING
POWER
SOURCE
.
37
TESTING
METHANOL
AS
A
REDUCING
POWER
SOURCE
.
38
CHAPTER
3:
ESTABLISHING
THE
SERINE
THREONINE
CYCLE
IN
COLL
.
.
43
PRODUCTION
OF
ACETYL-COA,
GLYCINE
AND
SERINE
VIA
THREONINE
SYNTHESIS
AND
CLEAVAGE
.
45
ESTABLISHMENT
OF
AN
EXTERNAL
ENERGY
PRODUCTION
MODULE
IMPROVES
GROWTH
ON
GLYCEROL
AND
ALLOWS
GROWTH
ON
PYRUVATE
.
49
FORMATE
IS
ASSIMILATED
TO
ALLOW
GROWTH
VIA
THE
SERINE
THREONINE
CYCLE
.
51
SHORT-TERM
EVOLUTION
IMPROVES
GROWTH
ON
PYRUVATE
AND
FORMATE
AND
ENABLES
GROWTH
ON
GLYCINE
AND
FORMATE
.
54
CHAPTER
4:
GROWTH
OF
E.
COU
ON
FORMATE
AND
METHANOL
VIA
THE
REDUCTIVE
GLYCINE
PATHWAY
.
57
THE
REDUCTIVE
GLYCINE
PATHWAY
.
59
MODULAR
ENGINEERING
ESTABLISHES
GROWTH
ON
FORMATE
.
61
SHORT-TERM
EVOLUTION
IMPROVES
GROWTH
ON
FORMATE
.
64
CARBON
LABELLING
SHEDS
LIGHT
ON
CELLULAR
FLUXES
.
66
ENGINEERED
GROWTH
OF
E.
COLI
ON
METHANOL
.
67
DISCUSSION
.
71
SYNTHETIC
BACTERIAL
METABOLISM
.
71
THE
POWER
OF
ADAPTIVE
LABORATORY
EVOLUTION
.
72
MEETING
CELLULAR
ENERGY
DEMANDS
BY
CI
OXIDATION
.
73
FORMATOTROPHIC
GROWTH
VIA
THE
REDUCTIVE
GLYCINE
PATHWAY
.
74
FORMATOTROPHIC
GROWTH
VIA
THE
SERINE
THREONINE
CYCLE
.
75
OUTLOOK
-
THE
FUTURE
OF
SYNTHETIC
CI
METABOLISM
.
77
APPENDIX
.
79
SUPPLEMENTARY
FIGURES
.
79
REFERENCES
.
89
DISCLAIMER
.
103
DECLARATION
OF
WORK
.
105
EIDESSTATTLICHE
ERKLARUNG
.
105
ACKNOWLEDGEMENTS
.
107
II |
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| id | DE-604.BV047334836 |
| illustrated | Illustrated |
| index_date | 2024-07-03T17:32:15Z |
| indexdate | 2024-07-10T09:09:17Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-032737397 |
| oclc_num | 1225838186 |
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| physical | V, 107 Seiten Illustrationen 30 cm |
| publishDate | 2020 |
| publishDateSearch | 2020 |
| publishDateSort | 2020 |
| record_format | marc |
| spelling | Wenk, Sebastian Verfasser (DE-588)1220117633 aut Engineering formatotrophic growth in Escherichia coli Sebastian Wenk Potsdam [2020] V, 107 Seiten Illustrationen 30 cm txt rdacontent n rdamedia nc rdacarrier Dissertation Universität Potsdam 2020 (DE-588)4113937-9 Hochschulschrift gnd-content B:DE-101 application/pdf https://d-nb.info/1223182495/04 Inhaltsverzeichnis DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032737397&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Wenk, Sebastian Engineering formatotrophic growth in Escherichia coli |
| subject_GND | (DE-588)4113937-9 |
| title | Engineering formatotrophic growth in Escherichia coli |
| title_auth | Engineering formatotrophic growth in Escherichia coli |
| title_exact_search | Engineering formatotrophic growth in Escherichia coli |
| title_exact_search_txtP | Engineering formatotrophic growth in Escherichia coli |
| title_full | Engineering formatotrophic growth in Escherichia coli Sebastian Wenk |
| title_fullStr | Engineering formatotrophic growth in Escherichia coli Sebastian Wenk |
| title_full_unstemmed | Engineering formatotrophic growth in Escherichia coli Sebastian Wenk |
| title_short | Engineering formatotrophic growth in Escherichia coli |
| title_sort | engineering formatotrophic growth in escherichia coli |
| topic_facet | Hochschulschrift |
| url | https://d-nb.info/1223182495/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032737397&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT wenksebastian engineeringformatotrophicgrowthinescherichiacoli |
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