Structure elucidation and biosynthesis of secondary metabolites containing non-proteinogenic amino acids from Xanthomonas and Streptomyces species
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
Berlin
2017
|
| Schlagworte: | |
| Online-Zugang: | Abstract Inhaltsverzeichnis Inhaltsverzeichnis |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
MARC
| LEADER | 00000nam a2200000 c 4500 | ||
|---|---|---|---|
| 001 | BV044329711 | ||
| 003 | DE-604 | ||
| 005 | 00000000000000.0 | ||
| 007 | t| | ||
| 008 | 170529s2017 xx a||| m||| 00||| eng d | ||
| 035 | |a (OCoLC)1001953987 | ||
| 035 | |a (DE-599)BVBBV044329711 | ||
| 040 | |a DE-604 |b ger |e rda | ||
| 041 | 0 | |a eng | |
| 049 | |a DE-83 | ||
| 100 | 1 | |a Saleh, Hesham |e Verfasser |0 (DE-588)1133051065 |4 aut | |
| 245 | 1 | 0 | |a Structure elucidation and biosynthesis of secondary metabolites containing non-proteinogenic amino acids from Xanthomonas and Streptomyces species |c vorgelegt von M.Sc. Hesham Saleh, geb. in Kairo, Ägypten |
| 264 | 1 | |a Berlin |c 2017 | |
| 300 | |a vii, 131 Seiten |b Illustrationen, Diagramme | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 502 | |b Dissertation |c Technische Universität Berlin |d 2017 | ||
| 655 | 7 | |0 (DE-588)4113937-9 |a Hochschulschrift |2 gnd-content | |
| 856 | 4 | |u http://verlag.tu-berlin.de/abstracts/diss/saleh_hesham.abstract.pdf |3 Abstract | |
| 856 | 4 | |u http://verlag.tu-berlin.de/abstracts/diss/saleh_hesham_inhalt.pdf |3 Inhaltsverzeichnis | |
| 856 | 4 | 2 | |m DNB Datenaustausch |q application/pdf |u http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=029732998&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |3 Inhaltsverzeichnis |
| 943 | 1 | |a oai:aleph.bib-bvb.de:BVB01-029732998 | |
Datensatz im Suchindex
| _version_ | 1819727519333482496 |
|---|---|
| adam_text | TABLE OF CONTENTS
ABBREVIATIONS.............................................................................................................................................I
TABLE OF
FIGURES........................................................................................................................................V
LIST OF
TABLES............................................................................................................................................VII
1
INTRODUCTION.................................................................................................................................1
1.1 DIVERSITY AND IMPACTS OF NATURAL
PRODUCTS........................................................................1
1.2 ADVANCEMENTS IN DISCOVERY OF LEAD-STRUCTURES FROM NATURAL PRODUCTS
..........................
6
1.2.1 TECHNOLOGICAL ADVANCEMENTS IN
BIOANALYTICS....................................................................6
1.2.2 BIOINFORMATICS ACCELERATE TARGET IDENTIFICATION
...............................................................
8
1.3 BIOSYNTHETIC ORIGINS FOR TERPENE AND PEPTIDE SECONDARY METABOLITES
.........................
10
1.3.1 TERPENES: THE PROBABLY LARGEST CLASS OF NATURAL
PRODUCTS.............................................12
1.3.2 NON-RIBOSOMALLY SYNTHESIZED PEPTIDES: STRUCTURAL AND BIOSYNTHETIC
COMPLEXITIES
....
13
1.4 SIGNIFICANCE OF BACTERIAL SECONDARY METABOLITES
...........................................................
18
1.4.1 THE GENUS
XANTHOMONAS: PHYTOPATHOGENS AIDING DRUG DISCOVERY
...............................
19
1.4.2 THE GENUS
STREPTOMYCES: THE PRINCIPAL PROVIDERS OF BIOACTIVE MOLECULES
...................
20
2 AIM OF THE W O
RK.......................................................................................................................
21
3 M
ATERIALS....................................................................................................................................
23
3.1 EQUIPMENT AND
SOFTWARE..................................................................................................23
3.2 CHEMICALS, ENZYMES AND
MATERIALS.................................................................................
26
3.3 PLASMIDS AND
OLIGONUCLEOTIDES........................................................................................
30
3.4 MICROORGANISMS AND
ANTIBIOTICS......................................................................................
31
3.4.1
MICROORGANISMS................................................................................................................31
3.4.2
ANTIBIOTICS...........................................................................................................................31
3.5 CULTURE
MEDIA....................................................................................................................32
3.6 KITS, BUFFERS AND
SOLUTIONS................................................................................................34
3.6.1
KITS.....................................................................................................................................
34
3.6.2
BUFFERS...............................................................................................................................
34
3.6.3
SOLUTIONS............................................................................................................................
35
4
METHODS.......................................................................................................................................
37
4.1 MICROBIOLOGICAL
METHODS..................................................................................................
37
4.1.1 CULTIVATION OF
STRAINS........................................................................................................
37
4.1.2 PRECURSOR AND ISOTOPE-LABELLED PRECURSOR FEEDING
........................................................
38
4.1.3 PREPARATION OF CACI2-COMPETENT 5. COLI DH5A CELLS
.........................................................
38
4.1.4 PREPARATION OF A STREPTOMYCES SPORE SUSPENSION
..........................................................
38
4.1.5 ISOLATION OF STREPTOMYCES M
UTANTS.................................................................................
38
4.1.6 AGAR DIFFUSION
ASSAYS........................................................................................................
39
4.2 ISOLATION AND PURIFICATION OF BACTERIAL METABOLITES
.........................................................
39
4.2.1 XAD EXTRACTION OF LIQUID XANTHOMONAS CULTURES
............................................................
39
4.2.2 SOLID-PHASE
EXTRACTION......................................................................................................
39
4.2.3 EXTRACTION OF SOLID STREPTOMYCES
CULTURES.......................................................................40
4.2.4 SIZE-EXCLUSION CHROMATOGRAPHY
......................................................................................
40
4.2.5 FLASH
CHROMATOGRAPHY......................................................................................................
40
4.2.6 PREPARATIVE
HPLC..............................................................................................................
41
4.2.7 SEMI-PREPARATIVE
HPLC.....................................................................................................
41
4.3 ANALYTICAL
METHODS............................................................................................................
42
4.3.1 ANALYTICAL HPLC
42
4.3.2 LC-(HR)-ESI-MSN
EXPERIMENTS..........................................................................................
42
4.3.3 MOLECULAR NETWORK
ANALYSIS............................................................................................
43
4.3.4 NMR
ANALYSIS....................................................................................................................
44
4.3.5 AMINO ACID ANALYSIS USING CHIRAL
GC-MS..........................................................................45
4.3.6 MALDI IMAGING
MS...........................................................................................................45
4.4
BIOINFORMATICS..................................................................................................................
47
4.4.1 GENOME
MINING................................................................................................................47
4.4.2 SEQUENCE ALIGNMENTS OF NRPS MODULAR
DOMAINS..........................................................47
4.4.3 MODELING OF
A-DOMAINS....................................................................................................48
4.5 BIOCHEMICAL
METHODS.......................................................................................................48
4.5.1 ATP-(32P)-PPJ EXCHANGE
ASSAY..........................................................................................
48
4.6 MOLECULAR BIOLOGY
METHODS............................................................................................
48
4.6.1 PCR AMPLIFICATION OF GENOMIC DNA SEGMENTS
................................................................
48
4.6.2 AGAROSE GEL
ELECTROPHORESIS............................................................................................
49
4.6.3 GENOMIC DNA, PCR PRODUCTS, PLASMID DNA PURIFICATION AND ANALYSIS
.........................
50
4.6.4 CLONING OF PLASMID INSERT INTO CACH-COMPETENT E. COLI DH5A CELLS
................................
51
4.6.5 PLASMID TRANSFORMATION INTO F. COLI ET12567/PUZ8002 CELLS BY
ELECTROPORATION
.......
52
4.6.6 CONJUGATION OF STREPTOMYCES SPORES WITH F. COLI ET12567/PUZ8002
CLONES
...............
52
5 RESULTS AND
DISCUSSION...........................................................................................................
55
5.1 DISCOVERY OF A PABA-CONTAINING METABOLITE FROM XANTHOMONAS CITRI PV.
MANGIFERAEINDICAE............................................................................................................
55
5.2 ISOLATION AND PURIFICATION OF THE TARGET
COMPOUND....................................................... 56
5.3 STRUCTURE ELUCIDATION OF THE PABA-CONTAINING M
ETABOLITE............................................59
5.4 CYTOTOXIC ACTIVITY OF XANTHOMONIC
ACID...........................................................................64
5.5 PRODUCTION OF STREPTOFACTINS BY STREPTOMYCES TENDAE TUE 901/8C
................................
68
5.6 PARTIAL AMINO ACID SEQUENCE OF STREPTOFACTIN B USING LC-(HR)-ESI-MS3
ANALYSIS
........
68
5.7
BIOINFORMATICS...................................................................................................................70
5.7.1 IDENTIFICATION OF THE PUTATIVE BIOSYNTHESIS GENE
CLUSTER...............................................70
5.7.2 SEQUENCE ALIGNMENTS OF THE NRPS
DOMAINS...................................................................73
5.7.3 SPECIFICITY-CONFERRING CODES OF THE ADENYLATION
DOMAINS..............................................73
5.8 SPECIFICITY OF THE A8 DOM
AIN............................................................................................75
5.9 MOLECULAR
BIOLOGY.............................................................................................................
77
5.9.1 PCR AMPLIFICATION AND LIGATION IN CONJUGATION PLASMIDS
................................................
77
5.9.2 CLONING OF PK18MOB_APRA-SFND INTO CACI2-COMPETENT E. COLI DH5A AND
E. COLI
ET12567/PUZ8002
CELLS.....................................................................................................79
5.9.3 CONJUGATION OF STREPTOMYCES SPORES WITH E. COLI ET12567/PUZ8002
CLONES................79
5.9.4 GENOMIC DNA ISOLATION AND
ANALYSIS...............................................................................
80
5.10 MALDI-IMS FOR THE PRODUCTION
STREPTOFACTINS................................................................81
5.11 ISOLATION AND PURIFICATION OF STREPTOFACTIN A
...................................................................82
5.12 STRUCTURE ELUCIDATION OF STREPTOFACTIN A
.........................................................................84
5.12.1 AMINO ACIDS ANALYSIS USING CHIRAL
GC-MS.....................................................................84
5.12.2 LC-(HR)-ESI-MS/MS
ANALYSIS.........................................................................................87
5.12.3 NMR
SPECTROSCOPY.......................................................................................................
90
5.13 BIOSYNTHESIS OF
STREPTOFACTINS..........................................................................................95
6 O
UTLOOK.......................................................................................................................................
99
7
APPENDIX...................................................................................................................................103
8
REFERENCES
123
|
| any_adam_object | 1 |
| author | Saleh, Hesham |
| author_GND | (DE-588)1133051065 |
| author_facet | Saleh, Hesham |
| author_role | aut |
| author_sort | Saleh, Hesham |
| author_variant | h s hs |
| building | Verbundindex |
| bvnumber | BV044329711 |
| ctrlnum | (OCoLC)1001953987 (DE-599)BVBBV044329711 |
| format | Thesis Book |
| fullrecord | <?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01407nam a2200301 c 4500</leader><controlfield tag="001">BV044329711</controlfield><controlfield tag="003">DE-604</controlfield><controlfield tag="005">00000000000000.0</controlfield><controlfield tag="007">t|</controlfield><controlfield tag="008">170529s2017 xx a||| m||| 00||| eng d</controlfield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(OCoLC)1001953987</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-599)BVBBV044329711</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-604</subfield><subfield code="b">ger</subfield><subfield code="e">rda</subfield></datafield><datafield tag="041" ind1="0" ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="049" ind1=" " ind2=" "><subfield code="a">DE-83</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Saleh, Hesham</subfield><subfield code="e">Verfasser</subfield><subfield code="0">(DE-588)1133051065</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Structure elucidation and biosynthesis of secondary metabolites containing non-proteinogenic amino acids from Xanthomonas and Streptomyces species</subfield><subfield code="c">vorgelegt von M.Sc. Hesham Saleh, geb. in Kairo, Ägypten</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="a">Berlin</subfield><subfield code="c">2017</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">vii, 131 Seiten</subfield><subfield code="b">Illustrationen, Diagramme</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="b">n</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="b">nc</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="502" ind1=" " ind2=" "><subfield code="b">Dissertation</subfield><subfield code="c">Technische Universität Berlin</subfield><subfield code="d">2017</subfield></datafield><datafield tag="655" ind1=" " ind2="7"><subfield code="0">(DE-588)4113937-9</subfield><subfield code="a">Hochschulschrift</subfield><subfield code="2">gnd-content</subfield></datafield><datafield tag="856" ind1="4" ind2=" "><subfield code="u">http://verlag.tu-berlin.de/abstracts/diss/saleh_hesham.abstract.pdf</subfield><subfield code="3">Abstract</subfield></datafield><datafield tag="856" ind1="4" ind2=" "><subfield code="u">http://verlag.tu-berlin.de/abstracts/diss/saleh_hesham_inhalt.pdf</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="856" ind1="4" ind2="2"><subfield code="m">DNB Datenaustausch</subfield><subfield code="q">application/pdf</subfield><subfield code="u">http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=029732998&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA</subfield><subfield code="3">Inhaltsverzeichnis</subfield></datafield><datafield tag="943" ind1="1" ind2=" "><subfield code="a">oai:aleph.bib-bvb.de:BVB01-029732998</subfield></datafield></record></collection> |
| genre | (DE-588)4113937-9 Hochschulschrift gnd-content |
| genre_facet | Hochschulschrift |
| id | DE-604.BV044329711 |
| illustrated | Illustrated |
| indexdate | 2024-12-24T05:57:29Z |
| institution | BVB |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-029732998 |
| oclc_num | 1001953987 |
| open_access_boolean | |
| owner | DE-83 |
| owner_facet | DE-83 |
| physical | vii, 131 Seiten Illustrationen, Diagramme |
| publishDate | 2017 |
| publishDateSearch | 2017 |
| publishDateSort | 2017 |
| record_format | marc |
| spellingShingle | Saleh, Hesham Structure elucidation and biosynthesis of secondary metabolites containing non-proteinogenic amino acids from Xanthomonas and Streptomyces species |
| subject_GND | (DE-588)4113937-9 |
| title | Structure elucidation and biosynthesis of secondary metabolites containing non-proteinogenic amino acids from Xanthomonas and Streptomyces species |
| title_auth | Structure elucidation and biosynthesis of secondary metabolites containing non-proteinogenic amino acids from Xanthomonas and Streptomyces species |
| title_exact_search | Structure elucidation and biosynthesis of secondary metabolites containing non-proteinogenic amino acids from Xanthomonas and Streptomyces species |
| title_full | Structure elucidation and biosynthesis of secondary metabolites containing non-proteinogenic amino acids from Xanthomonas and Streptomyces species vorgelegt von M.Sc. Hesham Saleh, geb. in Kairo, Ägypten |
| title_fullStr | Structure elucidation and biosynthesis of secondary metabolites containing non-proteinogenic amino acids from Xanthomonas and Streptomyces species vorgelegt von M.Sc. Hesham Saleh, geb. in Kairo, Ägypten |
| title_full_unstemmed | Structure elucidation and biosynthesis of secondary metabolites containing non-proteinogenic amino acids from Xanthomonas and Streptomyces species vorgelegt von M.Sc. Hesham Saleh, geb. in Kairo, Ägypten |
| title_short | Structure elucidation and biosynthesis of secondary metabolites containing non-proteinogenic amino acids from Xanthomonas and Streptomyces species |
| title_sort | structure elucidation and biosynthesis of secondary metabolites containing non proteinogenic amino acids from xanthomonas and streptomyces species |
| topic_facet | Hochschulschrift |
| url | http://verlag.tu-berlin.de/abstracts/diss/saleh_hesham.abstract.pdf http://verlag.tu-berlin.de/abstracts/diss/saleh_hesham_inhalt.pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=029732998&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT salehhesham structureelucidationandbiosynthesisofsecondarymetabolitescontainingnonproteinogenicaminoacidsfromxanthomonasandstreptomycesspecies |