Abstract
The enzymes of the tricarboxylic acid (TCA) and glyoxylate cycles of Corynebacterium glutamicum and in particular their regulation have been intensively studied in the past years. Nearly all TCA and glyoxylate cycle genes are subject to growth phase- or carbon source-dependent transcriptional regulation. Seven different regulators were shown to be involved in expression control of TCA and glyoxylate cycle genes, i.e., AcnR, DtxR, GlxR, RamA, RamB, RipA, and SucR. At the level of enzyme activity, the 2-oxoglutarate dehydrogenase (ODH) complex was found to be controlled by the inhibitor protein OdhI in dependency of its phosphorylation state, which is determined by the serine/threonine protein kinases PknG, PknA, PknB, and PknL and by the phospho-serine/threonine protein phosphatase Ppp. OdhI was shown to be crucial for glutamate production. This chapter summarizes new data on TCA cycle enzymes and describes the current knowledge on the regulation of this pathway and of the glyoxylate shunt.
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References
Arndt A, Eikmanns BJ (2008) Regulation of carbon metabolism in Corynebacterium glutamicum. In: Burkovski A (ed) Corynebacteria: genomics and molecular biology. Caister Academic, Norfolk, pp 155–182
Arndt A, Auchter M, Ishige T, Wendisch VF, Eikmanns BJ (2008) Ethanol catabolism in Corynebacterium glutamicum. J Mol Microbiol Biotechnol 15:222–233
Auchter M, Cramer A, Hüser A, Rückert C, Emer D, Schwarz P, Arndt A, Lange C, Kalinowski J, Wendisch VF, Eikmanns BJ (2010) RamA and RamB are global transcriptional regulators in Corynebacterium glutamicum and control genes for enzymes of the central metabolism. J Biotechnol 154:126–139
Barthe P, Roumestand C, Canova MJ, Kremer L, Hurard C, Molle V, Cohen-Gonsaud M (2009) Dynamic and structural characterization of a bacterial FHA protein reveals a new autoinhibition mechanism. Structure 17:568–578
Baumgart M, Bott M (2010) Biochemical characterisation of aconitase from Corynebacterium glutamicum. J Biotechnol 154:163–170
Baumgart M, Mustafi N, Bott M (2011) Deletion of the aconitase gene in Corynebacterium glutamicum causes a strong selection pressure for secondary mutations inactivating citrate synthase. J Bacteriol 193:6864–6873
Becker J, Klopprogge C, Schröder H, Wittmann C (2009) Metabolic engineering of the tricarboxylic acid cycle for improved lysine production by Corynebacterium glutamicum. Appl Environ Microbiol 75:7866–7869
Bendt AK, Burkovski A, Schaffer S, Bott M, Farwick M, Hermann T (2003) Towards a phosphoproteome map of Corynebacterium glutamicum. Proteomics 3:1637–1646
Bennett BD, Kimball EH, Gao M, Osterhout R, Van Dien SJ, Rabinowitz JD (2009) Absolute metabolite concentrations and implied enzyme active site occupancy in Escherichia coli. Nat Chem Biol 5:593–599
Bott M (2007) Offering surprises: TCA cycle regulation in Corynebacterium glutamicum. Trends Microbiol 15:417–425
Bott M (2010) Signal transduction by serine/threonine protein kinases in bacteria. In: Krämer R, Jung K (eds) Bacterial signaling. Wiley-VCH Verlag, Weinheim, pp 427–447
Bott M, Niebisch A (2003) The respiratory chain of Corynebacterium glutamicum. J Biotechnol 104:129–153
Brocker M, Schaffer S, Mack C, Bott M (2009) Citrate utilization by Corynebacterium glutamicum is controlled by the CitAB two-component system through positive regulation of the citrate transport genes citH and tctCBA. J Bacteriol 191:3869–3880
Brune I, Werner H, Hüser AT, Kalinowski J, Pühler A, Tauch A (2006) The DtxR protein acting as dual transcriptional regulator directs a global regulatory network involved in iron metabolism of Corynebacterium glutamicum. BMC Genomics 7:21
Bussmann M, Emer D, Hasenbein S, Degraf S, Eikmanns BJ, Bott M (2009) Transcriptional control of the succinate dehydrogenase operon sdhCAB of Corynebacterium glutamicum by the cAMP-dependent regulator GlxR and the LuxR-type regulator RamA. J Biotechnol 143:173–182
Cho H-Y, Lee SG, Han SO (2010) Identification and characterization of a transcriptional regulator, SucR, that influences sucCD transcription in Corynebacterium glutamicum. Biochem Biophys Res Commun 401:300–305
Claes WA, Pühler A, Kalinowski J (2002) Identification of two prpDBC gene clusters in Corynebacterium glutamicum and their involvement in propionate degradation via the 2-methylcitrate cycle. J Bacteriol 184:2728–2739
Cozzone AJ, El-Mansi M (2005) Control of isocitrate dehydrogenase catalytic activity by protein phosphorylation in Escherichia coli. J Mol Microbiol Biotechnol 9:132–146
Cramer A, Gerstmeir R, Schaffer S, Bott M, Eikmanns BJ (2006) Identification of RamA, a novel LuxR-type transcriptional regulator of genes involved in acetate metabolism of Corynebacterium glutamicum. J Bacteriol 188:2554–2567
Delaunay S, Gourdon P, Lapujade P, Mailly E, Oriol E, Engasser JM, Lindley ND, Goergen JL (1999) An improved temperature-triggered process for glutamate production with Corynebacterium glutamicum. Enzyme Microb Technol 25:762–768
Deutscher J, Saier MH Jr (2005) Ser/Thr/Tyr protein phosphorylation in bacteria—for long time neglected, now well established. J Mol Microbiol Biotechnol 9:125–131
Dietrich C, Nato A, Bost B, Le Maréchal P, Guyonvarch A (2009) Regulation of ldh expression during biotin-limited growth of Corynebacterium glutamicum. Microbiology 155:1360–1375
Ebbighausen H, Weil B, Krämer R (1991) Na+-dependent succinate uptake in Corynebacterium glutamicum. FEMS Microbiol Lett 77:61–65
Eikmanns BJ (2005) Central metabolism: tricarboxylic acid cycle and anaplerotic reactions. In: Eggeling L, Bott M (eds) Handbook of Corynebacterium glutamicum. CRC, Boca Raton, FL, pp 241–276
Eikmanns BJ, Thum-Schmitz N, Eggeling L, Lüdtke KU, Sahm H (1994) Nucleotide sequence, expression and transcriptional analysis of the Corynebacterium glutamicum gltA gene encoding citrate synthase. Microbiology 140:1817–1828
Eikmanns BJ, Rittmann D, Sahm H (1995) Cloning, sequence analysis, expression, and inactivation of the Corynebacterium glutamicum icd gene encoding isocitrate dehydrogenase and biochemical characterization of the enzyme. J Bacteriol 177:774–783
Emer D, Krug A, Eikmanns BJ, Bott M (2009) Complex expression control of the Corynebacterium glutamicum aconitase gene: Identification of RamA as a third transcriptional regulator besides AcnR and RipA. J Biotechnol 140:92–98
England P, Wehenkel A, Martins S, Hoos S, Andre-Leroux G, Villarino A, Alzari PM (2009) The FHA-containing protein GarA acts as a phosphorylation-dependent molecular switch in mycobacterial signaling. FEBS Lett 583:301–307
Fiuza M, Canova MJ, Zanella-Cleon I, Becchi M, Cozzone AJ, Mateos LM, Kremer L, Gil JA, Molle V (2008) From the characterization of the four serine/threonine protein kinases (PknA/B/G/L) of Corynebacterium glutamicum toward the role of PknA and PknB in cell division. J Biol Chem 283:18099–18112
Frunzke J, Bott M (2008) Regulation of iron homeostasis in Corynebacterium glutamicum. In: Burkovski A (ed) Corynebacteria: genomics and molecular Biology. Caister Academic, Norfolk, pp 241–266
Genda T, Watabe S, Ozaki H (2006) Purification and characterization of fumarase from Corynebacterium glutamicum. Biosci Biotechnol Biochem 70:1102–1109
Gerstmeir R, Wendisch VF, Schnicke S, Ruan H, Farwick M, Reinscheid D, Eikmanns BJ (2003) Acetate metabolism and its regulation in Corynebacterium glutamicum. J Biotechnol 104:99–122
Gerstmeir R, Cramer A, Dangel P, Schaffer S, Eikmanns BJ (2004) RamB, a novel transcriptional regulator of genes involved in acetate metabolism of Corynebacterium glutamicum. J Bacteriol 186:2798–2809
Hägerhall C, Hederstedt L (1996) A structural model for the membrane-integral domain of succinate:quinone oxidoreductases. FEBS Lett 389:25–31
Han SO, Inui M, Yukawa H (2008a) Effect of carbon source availability and growth phase on expression of Corynebacterium glutamicum genes involved in the tricarboxylic acid cycle and glyoxylate bypass. Microbiology 154:3073–3083
Han SO, Inui M, Yukawa H (2008b) Transcription of Corynebacterium glutamicum genes involved in tricarboxylic acid cycle and glyoxylate cycle. J Mol Microbiol Biotechnol 15:264–276
Hoffelder M, Raasch K, van Ooyen J, Eggeling L (2010) The E2 domain of OdhA of Corynebacterium glutamicum has succinyltransferase activity dependent on lipoyl residues of the acetyltransferase AceF. J Bacteriol 192:5203–5211
Huhn S, Jolkver E, Krämer R, Marin K (2010) Identification of the membrane protein SucE and its role in succinate transport in Corynebacterium glutamicum. Appl Microbiol Biotechnol 89:327–335
Inui M, Murakami S, Okino S, Kawaguchi H, Vertès AA, Yukawa H (2004) Metabolic analysis of Corynebacterium glutamicum during lactate and succinate productions under oxygen deprivation conditions. J Mol Microbiol Biotechnol 7:182–196
Inui M, Suda M, Okino S, Nonaka H, Puskas LG, Vertès AA, Yukawa H (2007) Transcriptional profiling of Corynebacterium glutamicum metabolism during organic acid production under oxygen deprivation conditions. Microbiology 153:2491–2504
Kalinowski J, Bathe B, Bartels D, Bischoff N, Bott M, Burkovski A, Dusch N, Eggeling L, Eikmanns BJ, Gaigalat L, Goesmann A, Hartmann M, Huthmacher K, Krämer R, Linke B, McHardy AC, Meyer F, Möckel B, Pfefferle W, Pühler A, Rey DA, Rückert C, Rupp O, Sahm H, Wendisch VF, Wiegrabe I, Tauch A (2003) The complete Corynebacterium glutamicum ATCC 13032 genome sequence and its impact on the production of L-aspartate-derived amino acids and vitamins. J Biotechnol 104:5–25
Kaspar S, Perozzo R, Reinelt S, Meyer M, Pfister K, Scapozza L, Bott M (1999) The periplasmic domain of the histidine autokinase CitA functions as a highly specific citrate receptor. Mol Microbiol 33:858–872
Kataoka M, Hashimoto KI, Yoshida M, Nakamatsu T, Horinouchi S, Kawasaki H (2006) Gene expression of Corynebacterium glutamicum in response to the conditions inducing glutamate overproduction. Lett Appl Microbiol 42:471–476
Kawahara Y, Takahashi-Fuke K, Shimizu E, Nakamatsu T, Nakamori S (1997) Relationship between the glutamate production and the activity of 2-oxoglutarate dehydrogenase in Brevibacterium lactofermentum. Biosci Biotechnol Biochem 61:1109–1112
Kim HJ, Kim TH, Kim Y, Lee HS (2004) Identification and characterization of glxR, a gene involved in regulation of glyoxylate bypass in Corynebacterium glutamicum. J Bacteriol 186:3453–3460
Kim J, Fukuda H, Hirasawa T, Nagahisa K, Nagai K, Wachi M, Shimizu H (2010) Requirement of de novo synthesis of the OdhI protein in penicillin-induced glutamate production by Corynebacterium glutamicum. Appl Microbiol Biotechnol 86:911–920
Kohl TA, Baumbach J, Jungwirth B, Pühler A, Tauch A (2008) The GlxR regulon of the amino acid producer Corynebacterium glutamicum: in silico and in vitro detection of DNA binding sites of a global transcription regulator. J Biotechnol 135:340–350
Krawczyk S, Raasch K, Schultz C, Hoffelder M, Eggeling L, Bott M (2010) The FHA domain of OdhI interacts with the carboxyterminal 2-oxoglutarate dehydrogenase domain of OdhA in Corynebacterium glutamicum. FEBS Lett 584:1463–1468
Krug A, Wendisch VF, Bott M (2005) Identification of AcnR, a TetR-type repressor of the aconitase gene acn in Corynebacterium glutamicum. J Biol Chem 280:585–595
Kurokawa T, Sakamoto J (2005) Purification and characterization of succinate:menaquinone oxidoreductase from Corynebacterium glutamicum. Arch Microbiol 183:317–324
Liang XY, Van Doren SR (2008) Mechanistic insights into phosphoprotein-binding FHA domains. Acc Chem Res 41:991–999
Lobell RB, Schleif RF (1990) DNA looping and unlooping by AraC protein. Science 250:528–532
Mahajan A, Yuan C, Lee H, Chen ESW, Wu PY, Tsai MD (2008) Structure and function of the phosphothreonine-specific FHA domain. Sci Signal 1(51):re12
Masse E, Gottesman S (2002) A small RNA regulates the expression of genes involved in iron metabolism in Escherichia coli. Proc Natl Acad Sci USA 99:4620–4625
Molenaar D, van der Rest ME, Petrovic S (1998) Biochemical and genetic characterization of the membrane-associated malate dehydrogenase (acceptor) from Corynebacterium glutamicum. Eur J Biochem 254:395–403
Molenaar D, van der Rest ME, Drysch A, Yücel R (2000) Functions of the membrane-associated and cytoplasmic malate dehydrogenases in the citric acid cycle of Corynebacterium glutamicum. J Bacteriol 182:6884–6891
Nakamura J, Hirano S, Ito H, Wachi M (2007) Mutations of the Corynebacterium glutamicum NCgl1221 gene, encoding a mechanosensitive channel homolog, induce L-glutamic acid production. Appl Environ Microbiol 73:4491–4498
Nesvera J, Patek M (2008) Plasmids and promoters in corynebacteria and their applications. In: Burkovski A (ed) Corynebacteria: genomics and molecular biology. Caister Academic, Norfolk, pp 113–154
Niebisch A, Kabus A, Schultz C, Weil B, Bott M (2006) Corynebacterial protein kinase G controls 2-oxoglutarate dehydrogenase activity via the phosphorylation status of the OdhI protein. J Biol Chem 281:12300–12307
Okino S, Noburyu R, Suda M, Jojima T, Inui M, Yukawa H (2008) An efficient succinic acid production process in a metabolically engineered Corynebacterium glutamicum strain. Appl Microbiol Biotechnol 81:459–464
Pallen M, Chuadhuri R, Khan A (2002) Bacterial FHA domains: neglected players in the phospho-threonine signalling game? Trends Microbiol 10:556–563
Perez J, Castaneda-Garcia A, Jenke-Kodama H, Müller R, Munoz-Dorado J (2008) Eukaryotic-like protein kinases in the prokaryotes and the myxobacterial kinome. Proc Natl Acad Sci USA 105:15950–15955
Polen T, Schluesener D, Poetsch A, Bott M, Wendisch VF (2007) Characterization of citrate utilization in Corynebacterium glutamicum by transcriptome and proteome analysis. FEMS Microbiol Lett 273:109–119
Radmacher E, Eggeling L (2007) The three tricarboxylate synthase activities of Corynebacterium glutamicum and increase of L-lysine synthesis. Appl Microbiol Biotechnol 76:587–595
Radmacher E, Stansen KC, Besra GS, Alderwick LJ, Maughan WN, Hollweg G, Sahm H, Wendisch VF, Eggeling L (2005) Ethambutol, a cell wall inhibitor of Mycobacterium tuberculosis, elicits L-glutamate efflux of Corynebacterium glutamicum. Microbiology 151:1359–1368
Ramos JL, Martinez-Bueno M, Molina-Henares AJ, Teran W, Watanabe K, Zhang XD, Gallegos MT, Brennan R, Tobes R (2005) The TetR family of transcriptional repressors. Microbiol Mol Biol Rev 69:326–356
Schaffer S, Burkovski A (2005) Proteomics. In: Eggeling L, Bott M (eds) Handbook of Corynebacterium glutamicum. CRC, Boca Raton, FL, pp 99–118
Schreiner ME, Fiur D, Holatko J, Patek M, Eikmanns B (2005) E1 enzyme of the pyruvate dehydrogenase complex in Corynebacterium glutamicum: molecular analysis of the gene and phylogenetic aspects. J Bacteriol 187:6005–6018
Schröder J, Tauch A (2010) Transcriptional regulation of gene expression in Corynebacterium glutamicum: the role of global, master and local regulators in the modular and hierarchical gene regulatory network. FEMS Microbiol Rev 34:685–737
Schultz C, Niebisch A, Gebel L, Bott M (2007) Glutamate production by Corynebacterium glutamicum: dependence on the oxoglutarate dehydrogenase inhibitor protein OdhI and protein kinase PknG. Appl Microbiol Biotechnol 76:691–700
Schultz C, Niebisch A, Schwaiger A, Viets U, Metzger S, Bramkamp M, Bott M (2009) Genetic and biochemical analysis of the serine/threonine protein kinases PknA, PknB, PknG and PknL of Corynebacterium glutamicum: evidence for non-essentiality and for phosphorylation of OdhI and FtsZ by multiple kinases. Mol Microbiol 74:724–741
Schwinde JW, Hertz PF, Sahm H, Eikmanns BJ, Guyonvarch A (2001) Lipoamide dehydrogenase from Corynebacterium glutamicum: molecular and physiological analysis of the lpd gene and characterization of the enzyme. Microbiology 147:2223–2231
Shah IM, Laaberki MH, Popham DL, Dworkin J (2008) A eukaryotic-like Ser/Thr kinase signals bacteria to exit dormancy in response to peptidoglycan fragments. Cell 135:486–496
Shiio I, Ozaki H (1970) Regulation of nicotinamide adenine dinucleotide phosphate-specific glutamate dehydrogenase from Brevibacterium flavum, a glutamate-producing bacterium. J Biochem 68:633–647
Shiio I, Ujigawatakeda K (1980) Presence and regulation of α-ketoglutarate dehydrogenase complex in a glutamate-producing bacterium, Brevibacterium flavum. Agric Biol Chem 44:1897–1904
Shiio I, Ozaki H, Mori M (1982) Glutamate metabolism in a gutamate-producing bacterium, Brevibacterium flavum. Agric Biol Chem 46:493–500
Shimizu H, Tanaka H, Nakato A, Nagahisa K, Kimura E, Shioya S (2003) Effects of the changes in enzyme activities on metabolic flux redistribution around the 2-oxoglutarate branch in glutamate production by Corynebacterium glutamicum. Bioprocess Biosyst Eng 25:291–298
Shingu H, Terui G (1971) Studies on process of glutamic acid fermentation at enzyme level. 1. Changes of α-ketoglutaric acid dehydrogenase in course of culture. J Ferment Technol 49:400–405
Shirai T, Nakato A, Izutani N, Nagahisa K, Shioya S, Kimura E, Kawarabayasi Y, Yamagishi A, Gojobori T, Shimizu H (2005) Comparative study of flux redistribution of metabolic pathway in glutamate production by two coryneform bacteria. Metab Eng 7:59–69
Stock JB, Ninfa AJ, Stock AM (1989) Protein phosphorylation and regulation of adaptive responses in bacteria. Microbiol Rev 53:450–490
Teramoto H, Shirai T, Inui M, Yukawa H (2008) Identification of a gene encoding a transporter essential for utilization of C4 dicarboxylates in Corynebacterium glutamicum. Appl Environ Microbiol 74:5290–5296
Usuda Y, Tujimoto N, Abe C, Asakura Y, Kimura E, Kawahara Y, Kurahashi O, Matsui H (1996) Molecular cloning of the Corynebacterium glutamicum (‘Brevibacterium lactofermentum’ AJ12036) odhA gene encoding a novel type of 2-oxoglutarate dehydrogenase. Microbiology 142:3347–3354
Uy D, Delaunay S, Goergen JL, Engasser JM (2005) Dynamics of glutamate synthesis and excretion fluxes in batch and continuous cultures of temperature-triggered Corynebacterium glutamicum. Bioprocess Biosyst Eng 27:153–162
van Ooyen J, Emer D, Bussmann M, Bott M, Eikmanns BJ, Eggeling L (2010) Citrate synthase in Corynebacterium glutamicum is encoded by two gltA transcripts which are controlled by RamA, RamB, and GlxR. J Biotechnol 154:140–148
van Ooyen J, Noack S, Bott M., Reth A, Eggeling L. (2012) Improved L-lysine production with Corynebacterium glutamicum and systemic insight into citrate synthase flux and activity. Biotechnol Bioeng. DOI 10.1002/bit.24486
Wendisch VF, Spies M, Reinscheid DJ, Schnicke S, Sahm H, Eikmanns BJ (1997) Regulation of acetate metabolism in Corynebacterium glutamicum: transcriptional control of the isocitrate lyase and malate synthase genes. Arch Microbiol 168:262–269
Wendisch VF, De Graaf AA, Sahm H, Eikmanns BJ (2000) Quantitative determination of metabolic fluxes during coutilization of two carbon sources: comparative analyses with Corynebacterium glutamicum during growth on acetate and/or glucose. J Bacteriol 182:3088–3096
Wennerhold J, Bott M (2006) The DtxR regulon of Corynebacterium glutamicum. J Bacteriol 188:2907–2918
Wennerhold J, Krug A, Bott M (2005) The AraC-type regulator RipA represses aconitase and other iron proteins from Corynebacterium under iron limitation and is itself repressed by DtxR. J Biol Chem 280:40500–40508
Yankovskaya V, Horsefield R, Tornroth S, Luna-Chavez C, Miyoshi H, Leger C, Byrne B, Cecchini G, Iwata S (2003) Architecture of succinate dehydrogenase and reactive oxygen species generation. Science 299:700–704
Yeats C, Finn RD, Bateman A (2002) The PASTA domain: a beta-lactam-binding domain. Trends Biochem Sci 27:438–440
Youn JW, Jolkver E, Krämer R, Marin K, Wendisch VF (2008) Identification and characterization of the dicarboxylate uptake system DccT in Corynebacterium glutamicum. J Bacteriol 190:6458–6466
Youn JW, Jolkver E, Krämer R, Marin K, Wendisch VF (2009) Characterization of the dicarboxylate transporter DctA in Corynebacterium glutamicum. J Bacteriol 191:5480–5488
Yu Z, Reichheld SE, Savchenko A, Parkinson J, Davidson AR (2010) A comprehensive analysis of structural and sequence conservation in the TetR family transcriptional regulators. J Mol Biol 400:847–864
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Bott, M., Eikmanns, B.J. (2013). TCA Cycle and Glyoxylate Shunt of Corynebacterium glutamicum . In: Yukawa, H., Inui, M. (eds) Corynebacterium glutamicum. Microbiology Monographs, vol 23. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-29857-8_10
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