and stereoselective hydroxylation of amino acids new fungal proline hydroxylases Regio

211 Pages·2016·10.72 MB·English

Regio- and stereoselective hydroxylation of
amino acids (cid:21) new fungal proline hydroxylases
Regio- und stereoselektive Hydroxylierung
von Aminos(cid:228)uren (cid:21) Neue pilzliche
Prolinhydroxylasen
INAUGURALDISSERTATION
zur Erlangung des Doktorgrades (Dr. rer. nat.)
an der Fakult(cid:228)t f(cid:252)r Chemie und Pharmazie
der Albert-Ludwigs-Universit(cid:228)t Freiburg
vorgelegt von
Dipl. Biologin Stefanie Houwaart
aus Jena
November 2016

Die vorliegende Arbeit entstand von Juni 2012 bis November 2016 am Lehrstuhl
f(cid:252)r Pharmazeutische und Medizinische Chemie des Instituts f(cid:252)r Pharmazeutische
WissenschaftenderAlbert-Ludwigs-Universit(cid:228)tFreiburgunterderAnleitungvonHerrn
Prof. Dr. Michael M(cid:252)ller.
Vorsitzender des Promotionsausschusses: Prof. Dr. Stefan Weber
Referent: Prof. Dr. Michael M(cid:252)ller
Koreferent: Prof. Dr. Andreas Bechthold
Drittpr(cid:252)fer: PD Dr. Erik Schleicher
Datum der m(cid:252)ndlichen Pr(cid:252)fung: 12.12.2016

Erkl(cid:228)rung
Hiermit erkl(cid:228)re ich, dass ich die vorliegende Arbeit selbstst(cid:228)ndig und nur unter
Verwendung der angegebenen Literatur und Hilfsmittel angefertigt sowie Zitate
kenntlich gemacht habe.

dedicated to my fabulous sister Andrea

Acknowledgments
This thesis is the product of a lot of people working hand in hand. I am very grateful to
Prof Dr Michael M(cid:252)ller for supervising my thesis and the great opportunity to
work in his group. I am especially thankful for scienti(cid:28)c and ethical discussions, the
great working conditions, helpful advise, and all of your support. I am also very
thankful for entrusting me with the interdisciplinary ethics seminar. Your motivation
for teaching students and connecting di(cid:27)erent academic disciplines is very inspirational.
Dr Wolfgang H(cid:252)ttel for the interesting interdisciplinary topic connecting biology
and chemistry. For your supervision, your ever open door for discussions, our friendly
working atmosphere, and the rigid proofreading of this manuscript.
Prof Dr Andreas Bechthold for co-supervising my thesis.
PD Dr Erik Schleicher for the EPR spectroscopy cooperation, fruitful discussions and
taking part in my thesis defence.
Kerstin Serrer in the working group of PD Dr Erik Schleicher for countless EPR
measurements and lots of discussion.
Prof Dr Oliver Einsle, Dr Stefan Gerhardt, Dr Daniel Wohlwend, Florian Kemper and
Antonio Espin for providing me with the lab equipment, guidance and practical help
for and during protein crystallization experiments.
Dr Alexander Fries for introducing me into mass spectrometry and supporting
the HPLC(cid:21)MS anaylsis of the tripeptides and their interpretation. Nadine Killius
for the synthesis of SNAC-l-proline and everything around and about that broad
topic. Alexandra Walter for her great support in the HPLC(cid:21)MS analysis of the
SNAC-l-proline assays. Christina Sommer-Kamann, Dr Katharina Gloria Hugentobler
and Alexandra Walter for their support on everything about organic chemistry. Sascha
Ferlaino for NMR measurements. Julian Haas, David Conradt, Dr Philippe Bisel,
Marcel Wilde and Nadine Killius for their great help interpreting NMR spectra. Dr
Jutta Siegrist for her helpful advice regarding HPLC. Dr Anja R(cid:252)ther for her help
with the tiny pH meter. Johanna Mattay for performing the seeding experiments with
HtyE. Dr Torsten Houwaart for informatic support.
Marita Wagner for her help and support in all aspects of lab life and for pleasant talks
outside of the lab. I will speci(cid:28)cally remember our poem and play during our institute
excursion in 2012.
I am very fortune to have worked with and learned from great scientists: Dr
Laura Mazzaferro, Dr Alexander Fries and Dr Eduard Frick. Thank you very much for
sharpening my way of scienti(cid:28)c thinking by also keeping the bigger scheme in view.
My diploma students Christina Sommer-Kamann and Ramona Kopton(cid:22)it was
a pleasure working with you. My bachelor students Michel Kather und Sonja Fros as
well as our students working in the lab Martin M(cid:252)ller and Thomas Schenk(cid:22)many
thanks for your work and help on the project, but most importantly for our great
working atmosphere.
My colleagues from the bureau and the lab for all the help, fruitful (non-)scienti(cid:28)c
discussions and fun: Dr Eduard Frick, Dr Laura Mazzaferro, Leon F(cid:252)rtges, Stefanie
Fromme, Johanna Mattay and Sebastian Obermaier. I am grateful to Christina
Sommer-Kamann who lightens every situation with her wit and charme for a lot of
laughter and support. Silja Mordhorst for scienti(cid:28)c and non-scienti(cid:28)c exchange. Dr
Katharina Gloria Hugentobler for bringing more color into my life. All members
of the polyacrylamide gel(cid:21)ice cream collaboration team for all the gels and our ice
cream breaks (I am sure I still owe you some!). All the girls of our noodles and
(knitting/crocheting) kneedles sessions. Karin Braun-Mai for our adventurous living
together. All the people in the working groups of Prof Dr Michael M(cid:252)ller, Dr Ste(cid:27)en
L(cid:252)deke, Jun.-Prof Jennifer Andexer and generally all of the OKH sta(cid:27) for the friendly
working atmosphere and all the help they provided, when needed.
Special thanks to our administrative sta(cid:27) for helping a lot with organising teaching
and personal logistics: Dr Mira Kuisle and Anja Semmler as well as Barbara Rees and
Marina Schwarzkopf.
Prof Dr Michael M(cid:252)ller and Dr Dominik Baltes for four years together as lecturer in
our interdisciplinary ethics seminar. It was a pleasure working with and learning from
you. Also many thanks to Dr Jonas Pavelka, Dr Michael Schwarz, Dr Katharina Gloria
Hugentobler and Johanna Mattay for your support in the lecturer team. An extra big
thank you to Dr Caroline F(cid:252)ndling for your guest talk and enriching our lectures with
your legal point of view.
Dr Jutta Siegrist, Dr Torsten Houwaart and Kerstin Serrer for intense proofreading of
this manuscript.