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Genetic control of carbohydrate uptake
and utilization in Corynebacterium
glutamicum




A dissertation submitted to the
Mathematisch-Naturwissenschaftliche Fakultät
of the Heinrich-Heine-Universität Düsseldorf



presented by
Verena Engels
born in Neuss



May 2008


The practical work of this thesis has been performed at the Institute of Biotechnology
1, Research Center Jülich, from July 2005 until April 2008 under the supervision of
Prof. Dr. V. F. Wendisch (University Münster).






Printed with the acceptance of the Mathematisch-Naturwissenschaftliche Fakultät of
the Heinrich-Heine-Universität Düsseldorf.






Examiner: Prof. Dr. Volker F. Wendisch
Institute of Molecular Microbiology and Biotechnology
Westfalian-Wilhelms-University Münster

Coexaminer: Prof. Dr. Michael Bott
Institute of Biotechnology 1
Research Center Jülich GmbH


Oral examination: June 25, 2008
Results described in this dissertation have been published in the following original
publications:

Engels, V. and Wendisch, V.F. (2007). The DeoR-type regulator SugR represses
expression of ptsG in Corynebacterium glutamicum. J Bacteriol 189: 2955-2966.

Frunzke, J., Engels, V., Hasenbein, S. Gätgens, C. and Bott, M. (2007). Co-
ordinated regulation of gluconate catabolism and glucose uptake in
Corynebacterium glutamicum by two functionally equivalent transcriptional
regulators, GntR1 and GntR2. Mol Microbiol 67: 305-322.

Engels, V. and Wendisch, V.F. (2008). The global repressor SugR controls
expression of glycolysis and pentose phosphate pathway genes in
Corynebacterium glutamicum. submitted to J Bacteriol

Georgi, T., Engels, V. and Wendisch, V.F. (2008). Regulation of L-lactate utilization
by the FadR-type regulator LldR of Corynebacterium glutamicum. J Bacteriol
190: 963-971

Engels, V. and Wendisch, V.F. (2008). sugR deletion improves L-lysine production in
Corynebacterium glutamicum. Appl Microbiol Biotechnol to be submitted

Engels, V. and Wendisch, V.F. (2008). Functional and biochemical characterization
of ScrB (Cg2927) as sucrose-6-phosphate hydrolase essential for sucrose
utilization by Corynebacterium glutamicum. submitted to FEMS Microbiol Lett


Patent application:

Wendisch, V.F., Engels, V., Eikmanns, B.J., Blombach, B., Bathe, B. and Thierbach,
G. (2007). Process for the fermentative preparation of organic chemical compounds
using coryneform bacteria in which the sugR gene is present in attenuated form.
US Appl. 60/960,375 (Filing date 27.09.2007) Content I
Content
1 Summary ………………………………………………………………………………………………………………. 1
2 Introduction …………………………………………………………………………………………………………. 2
2.1 Carbohydrate uptake by C. glutamicum ……………………………………………………..…….. 2
2.1.1 Glucose, fructose and sucrose uptake is mediated by the
phosphoenolpyruvate-dependent phosphotransferase system (PTS)
in C. glutamicum …………………………………………………………………………………………………... 3
2.1.2 Uptake of non-PTS carbohydrates like D-ribose or gluconate ……………………… 5
2.1.3 Uptake of the organic acids acetate and L-lactate by C. glutamicum ………….. 7
2.2 Central carbon metabolism pathways in C. glutamicum ………………………………... 9
2.3 Transcriptional regulators of carbon metabolism in C. glutamicum ……...…… 11
2.4 L-lysine biosynthesis in C. glutamicum …………………………………………………..……… 12
2.5 Aims of this work ………………………………………………………………………………………………… 15
3 Results ………………………………………………………………………………………………………………… 16
3.1 The DeoR-type regulator SugR represses expression of ptsG in
Corynebacterium glutamicum ………………………………………………………………………...… 18
3.2 Co-ordinated regulation of gluconate catabolism and glucose uptake
in Corynebacterium glutamicum by two functionally equivalent
transcriptional regulators, GntR1 and GntR2 ………………………………………………… 30
3.3 The global repressor SugR controls expression of glycolysis and
pentose phosphate pathway genes in Corynebacterium glutamicum …….…. 48
3.4 Regulation of L-lactate utilization by the FadR-type regulator LldR of
Corynebacterium glutamicum ………………………………………………………..…………………. 62
3.5 sugR deletion improves L-lysine production in Corynebacterium
glutamicum ……………………………………………………………………………………………..…………… 71
3.6 Functional and biochemical characterization of ScrB (Cg2927) as
sucrose-6-phosphate hydrolase essential for sucrose utilization by
Corynebacterium glutamicum …………………………...……………………………………………… 80
II Content
4 Discussion …………………………………………………………………………………………………………. 89
4.1 Regulation of carbon metabolism in C. glutamicum as compared
to other bacteria like E. coli and B. subtilis …………………………………………………….. 89
4.2 Regulation of L-lactate metabolism in C. glutamicum by the global
regulator SugR and the FadR-type regulator LldR ………………………….……………. 99
4.3 Improvement of L-lysine production with C. glutamicum …………..……………….. 104
4.4 Conclusions and Perspectives …………………………..…………………………………………… 110
5 References ……………………………………………………………………..………………………………… 112
6 Appendix ………………………………………………….………………………………………………….……. 124
Supplementary material - ScrB.
















Abbreviations III
Abbreviations

RAmp Ampicillin resistance
ATCC American Type Culture Collection
ATPase adenosine triphosphatase
ChIP Chromatin immunoprecipitation
DTT dithiothreitol
e.g. for example
et al. et alii
i.e. that is
IPTG Isopropyl-thio- β-D-galactopyranosid
RKan Kanamycin resistance
MES 2-(N-morpholino)-ethanesulfonic acid
MOPS 3-(N-morpholino)-propanesulfonic acid
ORF open reading frame
RBS ribosome binding site
rpm revolutions per minute
v/v volume per volume
WT wild type
ε molar extinction coefficient



Abbreviations not included in this section are according to international standards, as
for example listed in the author guidelines of the FEBS Journal.

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