Investigation on the biosynthesis of the MraY inhibitors caprazamycins and napsamycins/mureidomycins: = Untersuchungen zur Biosynthese der MraY Translokase Inhibitoren Caprazamycine und Napsamycine/Mureidomycine
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2013
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XIII, 122 S. Ill., graph. Darst. |
Internformat
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| 245 | 1 | 0 | |a Investigation on the biosynthesis of the MraY inhibitors caprazamycins and napsamycins/mureidomycins |b = Untersuchungen zur Biosynthese der MraY Translokase Inhibitoren Caprazamycine und Napsamycine/Mureidomycine |c vorgelegt von Xiaoyu Tang |
| 246 | 1 | 1 | |a Untersuchungen zur Biosynthese der MraY Translokase Inhibitoren Caprazamycine und Napsamycine/Mureidomycine |
| 264 | 1 | |c 2013 | |
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Datensatz im Suchindex
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|---|---|
| adam_text | CONTENTS
CONTENTS
PUBLICATIONS AND POSTERS IV
1. PUBLICATIONS IV
2. CONFERENCE CONTRIBUTIONS IV
ABBREVATIONS VI
SUMMARY X
ZUSAMMENFASSUNG XII
I. INTRODUCTION 1
1. PEPTIDOGLYCAN AND ANTIBIOTICS TARGET
ON TRANSLOCASE MRAY 1
2. BIOLOGICAL ACTIVITY OF UPAS 4
3. BIOSYNTHESIS OF THE PACIDAMYCIN/MUREIDOMYCIN FAMILY OF UPAS 4
4.
BIOSYNTHESIS OF THE CAPTAZAMYCIN/LIPOSIDOMYCIN FAMILY OF
UPAS 7
5. SULFATION IN LIVING ORGANISMS 12
6. AIMS OF THIS STUDY 14
II. MATERIALS AND METHODS 16
1. CHEMICALS 16
2. MATERIALS FOR CHROMATOGRAPHY 19
3. ENZYMES AND KITS 19
4. MEDIA, BUFFERS AND SOLUTIONS 20
4.1. MEDIA FOR BACTERIAL CULTIVATION 20
4.2. ANTIBIOTIC SOLUTIONS 24
4.3. BUFFERS AND SOLUTIONS 25
5. PLASMIDS, BACTERIAL STRAINS, PRIMERS AND PROBES 29
5.1. VECTORS, COSMIDS
AND PLASMIDS 29
5.2. BACTERIAL STRAINS 31
6. CULTURE CONDITIONS 33
6.1. CULTIVATION OF E. COLI 33
6.2. CULTIVATION AND PRESERVATION OF STREPTOMYCES 33
7. METHODS OF MOLECULAR BIOLOGY 34
7.1. PURIFICATION, CONCENTRATION AND QUANTIFICATION OF DNA 34
7.2. AGAROSE GEL ELECTROPHORESIS
OF DNA 35
7.3. DNA MANIPULATION WITH ENZYMES 35
7.4. DNA ISOLATION 35
7.5. PCR AMPLIFICATION 37
7.6. INTRODUCTION OF DNA IN E. COLI 38
7.7. PEG-MEDIATED PROTOPLAST TRANSFORMATION FOR INTRODUCTION OF DNA
PREPARATION OF PROTOPLASTS FROM STREPTOMYCES 39
7.8. INTEIGENERIC CONJUGATION FOR INTRODUCTION OF DNA IN STREPTOMYCES 41
7.9. GENERATION AND SCREENING
OF A COSMID LIBRARY 42
7.10. DNA SEQUENCING AND COMPUTER-ASSISTED
SEQUENCE ANALYSIS 43
8. METHODS OF BIOCHEMISTRY AND BIOLOGY : 44
8.1. DENATURING POLYACRYLAMIDE GEL ELECTROPHORESIS
(SDS-PAGE) 44
8.2. CLONING AND PURIFICATION OF RECOMBINANT PROTEINS FROM E. COLI 44
II
HTTP://D-NB.INFO/1043494804
CONTENTS
8.3. ASSAY CONDITIONS 45
9. CONSTRUCTION OF STREPTOMYCES MUTANT STRAINS 47
10. ANALYSIS AND ISOLATION OF SECONDARY METABOLITES 51
10.1. LC-MS ANALYSIS 51
10.2. EXTRACTION OF CAPRAZAMYCIN OR NAPSAMYCIN DERIVATIVES 51
10.3. PURIFICATION OF HYDROXYACYLCAPRAZOL E 52
11. PRODUCTION,
ISOLATION,
PURIFICATION, AND
STRUCTURE DETERMINATION
OF PRESULFICIDINS
A, B, C AND D 52
11.1. PRODUCTION OF PRESULFICIDINS 52
11.2. PURIFICATION OF PRESULFICIDINS 53
11.3. NMR METHODS AND STRUCTURAL CHARACTERIZATION 53
III. RESULTS AND
DISCUSSION 55
1. INVESTIGATION ON NAPSAMYCIN BIOSYNTHESIS 55
1.1. IDENTIFICATION AND CLONING OF THE NAPSAMYCIN
GENE CLUSTER 55
1.2. SEQUENCE ANALYSIS
OF THE NAPSAMYCIN GENE
CLUSTER 56
1.3. HETEROLOGOUS EXPRESSION OF THE NAPSAMYCIN GENE CLUSTER 63
1.4. ANALYSIS OF THE NPSU KNOCK-OUT MUTANT 67
1.5. IDENTIFICATION OF MUREIDOMYCIN ANALOGUES 69
1.6. IDENTIFICATION OF N-ACETYLATED MUREIDOMYCIN B REVEALS A PLAUSIBLE
SELF-RESISTANT MECHANISM 73
1.7. CONCLUSIONS 78
2. A TWO-STEP
SULFATION MECHANISM IN CAPRAZAMYCIN BIOSYNTHESIS 80
2.1. ANALYSIS OF DELETION MUTANTS IN S.
COELICOLOR M512 81
2.2. FUNCTIONAL ASSIGNMENT OF CPZ8
AS A
PAPS-DEPENDENT SULFOTRANSFERASE 81
2.3. VALIDATION OF CPZ7 AS A PAP PHOSPHATASE 85
2.4. CPZ6 IS ESSENTIAL FOR THE SULFATION OF CAPRAZAMYCINS 87
2.5. PRESULFICIDINS ARE THE
PRODUCTS OF THE
TYPE III PKS
CPZ6 90
2.6. TWO-STEP SULFATION MECHANISM 94
2.7. CONCULSIONS 99
IV. REFERENCES 101
V. ACKNOWLEDGMENT 108
VI. CURRICULUM VITAE 109
VH. APPENDIX 110
111
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| any_adam_object | 1 |
| author | Tang, Xiaoyu 1984- |
| author_GND | (DE-588)1037989694 |
| author_facet | Tang, Xiaoyu 1984- |
| author_role | aut |
| author_sort | Tang, Xiaoyu 1984- |
| author_variant | x t xt |
| building | Verbundindex |
| bvnumber | BV041376408 |
| ctrlnum | (OCoLC)870907414 (DE-599)BVBBV041376408 |
| dewey-full | 540 610 |
| dewey-hundreds | 500 - Natural sciences and mathematics 600 - Technology (Applied sciences) |
| dewey-ones | 540 - Chemistry and allied sciences 610 - Medicine and health |
| dewey-raw | 540 610 |
| dewey-search | 540 610 |
| dewey-sort | 3540 |
| dewey-tens | 540 - Chemistry and allied sciences 610 - Medicine and health |
| discipline | Chemie / Pharmazie Medizin |
| format | Thesis Book |
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| indexdate | 2024-07-10T00:55:19Z |
| institution | BVB |
| language | English |
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| physical | XIII, 122 S. Ill., graph. Darst. |
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| spelling | Tang, Xiaoyu 1984- Verfasser (DE-588)1037989694 aut Investigation on the biosynthesis of the MraY inhibitors caprazamycins and napsamycins/mureidomycins = Untersuchungen zur Biosynthese der MraY Translokase Inhibitoren Caprazamycine und Napsamycine/Mureidomycine vorgelegt von Xiaoyu Tang Untersuchungen zur Biosynthese der MraY Translokase Inhibitoren Caprazamycine und Napsamycine/Mureidomycine 2013 XIII, 122 S. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Tübingen, Univ., Diss., 2013 Translocasen (DE-588)4760612-5 gnd rswk-swf Inhibitor (DE-588)4027007-5 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content Translocasen (DE-588)4760612-5 s Inhibitor (DE-588)4027007-5 s DE-604 DNB Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=026824460&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Tang, Xiaoyu 1984- Investigation on the biosynthesis of the MraY inhibitors caprazamycins and napsamycins/mureidomycins = Untersuchungen zur Biosynthese der MraY Translokase Inhibitoren Caprazamycine und Napsamycine/Mureidomycine Translocasen (DE-588)4760612-5 gnd Inhibitor (DE-588)4027007-5 gnd |
| subject_GND | (DE-588)4760612-5 (DE-588)4027007-5 (DE-588)4113937-9 |
| title | Investigation on the biosynthesis of the MraY inhibitors caprazamycins and napsamycins/mureidomycins = Untersuchungen zur Biosynthese der MraY Translokase Inhibitoren Caprazamycine und Napsamycine/Mureidomycine |
| title_alt | Untersuchungen zur Biosynthese der MraY Translokase Inhibitoren Caprazamycine und Napsamycine/Mureidomycine |
| title_auth | Investigation on the biosynthesis of the MraY inhibitors caprazamycins and napsamycins/mureidomycins = Untersuchungen zur Biosynthese der MraY Translokase Inhibitoren Caprazamycine und Napsamycine/Mureidomycine |
| title_exact_search | Investigation on the biosynthesis of the MraY inhibitors caprazamycins and napsamycins/mureidomycins = Untersuchungen zur Biosynthese der MraY Translokase Inhibitoren Caprazamycine und Napsamycine/Mureidomycine |
| title_full | Investigation on the biosynthesis of the MraY inhibitors caprazamycins and napsamycins/mureidomycins = Untersuchungen zur Biosynthese der MraY Translokase Inhibitoren Caprazamycine und Napsamycine/Mureidomycine vorgelegt von Xiaoyu Tang |
| title_fullStr | Investigation on the biosynthesis of the MraY inhibitors caprazamycins and napsamycins/mureidomycins = Untersuchungen zur Biosynthese der MraY Translokase Inhibitoren Caprazamycine und Napsamycine/Mureidomycine vorgelegt von Xiaoyu Tang |
| title_full_unstemmed | Investigation on the biosynthesis of the MraY inhibitors caprazamycins and napsamycins/mureidomycins = Untersuchungen zur Biosynthese der MraY Translokase Inhibitoren Caprazamycine und Napsamycine/Mureidomycine vorgelegt von Xiaoyu Tang |
| title_short | Investigation on the biosynthesis of the MraY inhibitors caprazamycins and napsamycins/mureidomycins |
| title_sort | investigation on the biosynthesis of the mray inhibitors caprazamycins and napsamycins mureidomycins untersuchungen zur biosynthese der mray translokase inhibitoren caprazamycine und napsamycine mureidomycine |
| title_sub | = Untersuchungen zur Biosynthese der MraY Translokase Inhibitoren Caprazamycine und Napsamycine/Mureidomycine |
| topic | Translocasen (DE-588)4760612-5 gnd Inhibitor (DE-588)4027007-5 gnd |
| topic_facet | Translocasen Inhibitor Hochschulschrift |
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