Tailor-made sortase reactions:
Gespeichert in:
| Beteilige Person: | |
|---|---|
| Format: | Hochschulschrift/Dissertation Buch |
| Sprache: | Englisch |
| Veröffentlicht: |
Aachen
[2018?]
|
| Schlagwörter: | |
| Links: | http://d-nb.info/1186906928/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=031426981&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| Umfang: | 172 Blätter Illustrationen 30 cm |
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| 100 | 1 | |a Schatte, Martin |e Verfasser |0 (DE-588)1185293957 |4 aut | |
| 245 | 1 | 0 | |a Tailor-made sortase reactions |c vorgelegt von Master of Science (M. Sc.) Biochemie Martin Schatte |
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Datensatz im Suchindex
| _version_ | 1819295405995720704 |
|---|---|
| adam_text | TABLE
OF
CONTENT
ABBREVIATIONS
AND
ACRONYMS
10
GRAPHICAL
ABSTRACT
12
ABSTRACT
13
INTRODUCTION
SORTASES
15
THE
NATIVE
FUNCTION
OF
SORTASES
A
15
SORTASE
REACTION
MECHANISM
16
SORTASE
CLASSES
AND
THEIR
FUNCTION
18
SORTASE
STRUCTURE
19
STRUCTURE
SORTASE
CLASS
A
21
STRUCTURE
SA-SRTA
21
STRUCTURE
SORTASE
CLASSES
C
AND
B
21
BIOCONJUGATION
22
ENZYMATIC
BIOCONJUGATION
22
LIGATION-TAGS
FOR BIOCONJUGATION
23
CHEMICAL
BIOCONJUGATION
24
NATIVE
CHEMICAL
LIGATION
25
MATERIALS
AND
METHODS
REAGENTS
26
BUFFER
29
KITS
30
4
PRIMER
30
INSTRUMENTS
35
E.COLI
STRAINS
37
E.COLI
PLASMIDS
37
SORTASE
CONSTRUCTS
37
METHODS
39
SOLID-PHASE
PEPTIDE
SYNTHESIS
OF
SORTASE
SUBSTRATES
39
SORTASE
EXPRESSION
AND
PURIFICATION
39
FRET
BASED
ASSAY
39
GLUCOSE
DEHYDROGENASE
(PQQ-GDH)
39
POLYACRYLAMIDE
GEL
ELECTROPHORESIS
(SDS-PAGE)
40
RE
LA
AS
SORTASE
ACTIVITY
ASSAY
40
REIA
AS
HIGH-THROUGHPUT
SCREENING
ASSAY
IN
CRUDE
CELLULAR EXTRACT
41
EVALUATION
OF
THE
BINDING
BEHAVIOR
ON
THE
STREPTAVIDIN
COATED
MTP
41
P450
EXPRESSION
41
CO-BINDING
ASSAY
42
PNCA
ASSAY
42
BCCE
ASSAY
42
SORTASE
ACTIVITY
ASSAY
42
WESTERN
BLOT
43
HPLC
MS
43
FORMATION
OF
DES
43
REACTION
IN
DIFFERENT
SOLVENTS
43
SOLUBILITY
OF
LIPOPHILIC
SUBSTRATES
44
5
INFLUENCE
OF
WATER
CONTENT
44
COUPLING
OF
TWO
LIPOPHILIC
SUBSTRATES
44
PLASMID
DNA
PREPARATION
45
ELECTROPORATION
45
RELATIVE
AFFINITY
QUOTIENT
(RAQ)
AND
RELATIVE
STABILITY
QUOTIENT
(RSQ)
DETERMINATION
45
DIFFERENTIAL
SCANNING
CALORIMETRY
45
DM
SIMULATIONS
45
A
REPORTER
IMMOBILIZATION
ASSAY
(REIA)
FOR
BIOCONJUGATING
REACTIONS
INTRODUCTION
47
RESULTS
AND
DISCUSSION
FRET-BASED
BIOCONJUGATION
ASSAY
49
REPORTER
IMMOBILIZATION
ASSAY
(REIA)
53
REAGENT
GENERATION
AND
EVALUATION
54
BIOTIN
AS
AFFINITY
HANDLE
56
ASSAY
OPTIMIZATION
58
FINAL
ASSAY PROTOCOL
60
REIA
AS
SCREENING
ASSAY
60
MULTIPARAMETER
SCREENING
OF
SORTASE
REACTIONS
USING
REIA
61
ULTRA-SENSITIVE
REPORTER
IMMOBILIZATION
ASSAY
(US-RELA)
63
WORKING
PROTOCOL
FOR
US-REIA
65
REIA
FOR
ANALYZING
GENERAL
BIOCONJUGATING
REACTIONS
68
6
ADOPTING
THE
REIA
TO
FLUOROPHORES
AS
REPORTER
69
CONCLUSION
71
PRESCREENING
P450
SPLIT
ENZYME
INTRODUCTION
72
RESULTS
AND
DISCUSSION
74
CONCLUSION
80
ENHANCED
SORTASE
BIOCONJUGATION
YIELD
INTRODUCTION
81
RESULTS
AND
DISCUSSION
BLOCKING
THE
BACK
REACTION
WITH
NONSTANDARD
ALA
OR CYS-NUCLEOPHILES
83
SORTASE
CATALYZED
NATIVE
CHEMICAL
LIGATION
(SRT-NCL)
87
CONCLUSION
95
DES
ENABLES
SORTASE
CONJUGATION
OF
HYDROPHOBIC
COMPOUNDS
INTRODUCTION
97
RESULTS
AND
DISCUSSION
IDENTIFICATION
OF
A
SUITABLE
SOLVENT
SYSTEM
FOR
SA-SRTA
100
INFLUENCE
OF
WATER
ON
SRTA
PERFORMANCE
IN
DES
(CHCEGLYCEROL
1
:2)
103
SOLUBILITY
OF
LIPOPHILIC
SUBSTRATES
IN
CHCEGLYCEROL
BASED
DEEP
EUTECTIC
SOLVENTS
105
COUPLING
OF
TWO
HYDROPHOBIC
SUBSTRATES
WITH
SA-SRTA
107
GLYCEROL
ACTS
AS
NUCLEOPHILE
IN
THE
DEEP
EUTECTIC
SOLVENT
108
(CHCEGLYCEROL
1:2)
7
CONCLUSION
110
ANALYZING
THE
NATURAL
DIVERSITY
OF
SORTASES
INTRODUCTION
112
RESULTS
GENETIC
CONSTRUCTIONS
OF
SORTASES
FOR
IN
VITRO
USE
114
METHOD
TO
GENERATE
SOLUBLE
AND
CATALYTICALLY
ACTIVE
SORTASES
115
SORTASE
EXPRESSION
AND
PURIFICATION
116
SORTASE
STABILITY
118
SORTASE
ACTIVITY
119
SORTASE
YIELDS
120
SUBSTRATE
CONCENTRATION
DEPENDENCY
123
CONCLUSIONS
125
SEMI-RATIONAL
DESIGN
OF
SORTASES
INTRODUCTION
PROTEIN
ENGINEERING
127
DIRECTED
EVOLUTION,
RATIONAL
DESIGN
AND
SEMI
RATIONAL
DESIGN
127
GENERATION
OF
MUTATIONS
128
SCREENING
ASSAY/SYSTEM
129
SORTASE
129
RESULTS
AND
DISCUSSION
RESULTS
OF
THE
FIRST
ROUND
OF
DIRECTED
EVOLUTION
USING
OMNICHANGE
131
RESULTS
OF
THE
SECOND
ROUND
OF
DIRECTED
EVOLUTION
USING
SESAM
135
RESULTS
OF
THE
THIRD
ROUND
OF
DIRECTED
EVOLUTION
138
8
CHARACTERIZATION
OF
THE
FINAL
MS
5
MUTANT
139
CONCLUSION
141
STRUCTURE-FUNCTION
RELATIONSHIP
OF
SA-SRTA:
MOLECULAR
BASIS
OF
THE
ACTIVITY
INCREASE
OF
MUTATED
SORTASE
2
(MS2)
INTRODUCTION
143
RESULTS
AND
DISCUSSION
STARTING
STRUCTURE
149
MD-SIMULATIONS
151
MD-SIMULATIONS
OF
WT
VS.
MS2
IN
*THR
OUT
*
STATE
153
CONCLUSION
158
SUMMARY
159
ZUSAMMENFASSUNG
162
PARTS
OF
THIS
THESIS
HAVE
BEEN
PUBLISHED
PREVIOUSLY
163
REFERENCES
164
9
|
| any_adam_object | 1 |
| author | Schatte, Martin |
| author_GND | (DE-588)1185293957 |
| author_facet | Schatte, Martin |
| author_role | aut |
| author_sort | Schatte, Martin |
| author_variant | m s ms |
| building | Verbundindex |
| bvnumber | BV046045362 |
| ctrlnum | (OCoLC)1108926817 (DE-599)DNB1186906928 |
| discipline | Chemie / Pharmazie Biologie |
| format | Thesis Book |
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| illustrated | Illustrated |
| indexdate | 2024-12-20T18:41:43Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-031426981 |
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| owner_facet | DE-355 DE-BY-UBR |
| physical | 172 Blätter Illustrationen 30 cm |
| publishDate | 2018 |
| publishDateSearch | 2018 |
| publishDateSort | 2018 |
| record_format | marc |
| spellingShingle | Schatte, Martin Tailor-made sortase reactions |
| subject_GND | (DE-588)4113937-9 |
| title | Tailor-made sortase reactions |
| title_auth | Tailor-made sortase reactions |
| title_exact_search | Tailor-made sortase reactions |
| title_full | Tailor-made sortase reactions vorgelegt von Master of Science (M. Sc.) Biochemie Martin Schatte |
| title_fullStr | Tailor-made sortase reactions vorgelegt von Master of Science (M. Sc.) Biochemie Martin Schatte |
| title_full_unstemmed | Tailor-made sortase reactions vorgelegt von Master of Science (M. Sc.) Biochemie Martin Schatte |
| title_short | Tailor-made sortase reactions |
| title_sort | tailor made sortase reactions |
| topic_facet | Hochschulschrift |
| url | http://d-nb.info/1186906928/04 http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=031426981&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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