Browse > Article
http://dx.doi.org/10.4014/jmb.1310.10031

Elucidation of the Regulation of Ethanol Catabolic Genes and ptsG Using a glxR and Adenylate Cyclase Gene (cyaB) Deletion Mutants of Corynebacterium glutamicum ATCC 13032  

Subhadra, Bindu (Department of Biomedicinal Science and Biotechnology, Paichai University)
Lee, Jung-Kee (Department of Biomedicinal Science and Biotechnology, Paichai University)
Publication Information
Journal of Microbiology and Biotechnology / v.23, no.12, 2013 , pp. 1683-1690 More about this Journal
Abstract
The cyclic AMP receptor protein (CRP) homolog, GlxR, controls the expression of several genes involved in the regulation of diverse physiological processes in Corynebacterium glutamicum. In silico analysis has revealed the presence of glxR binding sites upstream of genes ptsG, adhA, and ald, encoding glucose-specific phosphotransferase system protein, alcohol dehydrogenase (ADH), and acetaldehyde dehydrogenase (ALDH), respectively. However, the involvement of the GlxR-cAMP complex on the expression of these genes has been explored only in vitro. In this study, the expressions of ptsG, adhA, and ald were analyzed in detail using an adenylate cyclase gene (cyaB) deletion mutant and glxR deletion mutant. The specific activities of ADH and ALDH were increased in both the mutants in glucose and glucose plus ethanol media, in contrast to the wild type. In accordance, the promoter activities of adhA and ald were derepressed in the cyaB mutant, indicating that glxR acts as a repressor of adhA. Similarly, both the mutants exhibited derepression of ptsG regardless of the carbon source. These results confirm the involvement of GlxR on the expression of important carbon metabolic genes; adhA, ald, and ptsG.
Keywords
Corynebacterium glutamicum; GlxR; cyaB; adhA; ald; ptsG;
Citations & Related Records
연도 인용수 순위
  • Reference
1 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.   DOI   ScienceOn
2 Kohl TA, Baumbach J, Jungwirth B, Puhler 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.   DOI   ScienceOn
3 Kohl TA, Tauch A. 2009. The GlxR regulon of the amino acid producer Corynebacterium glutamicum: detection of the corynebacterial core regulon and integration into the transcriptional regulatory network model. J. Biotechnol. 143: 239-246.   DOI   ScienceOn
4 Kotrbova-Kozak A, Kotrba P, Inui M, Sajdok J, Yukawa H. 2007. Transcriptionally regulated adhA gene encodes alcohol dehydrogenase required for ethanol and n-propanol utilization in Corynebacterium glutamicum R. Appl. Microbiol. Biotechnol. 76: 1347-1356.   DOI   ScienceOn
5 Lee HW, Pan JG, Lebeault JM. 1998. Enhanced l-lysine production in threonine-limited continuous culture of Corynebacterium glutamicum by using gluconate as a secondary carbon source with glucose. Appl. Microbiol. Biotechnol. 49: 9-15.   DOI
6 Merkens H, Beckers G, Wirtz A, Burkovski A. 2005. Vanillate metabolism in Corynebacterium glutamicum. Curr. Microbiol. 51: 59-65.   DOI   ScienceOn
7 Mori A, Shiio I. 1987. Phosphoenol pyruvate: sugar phosphotransferase systems and sugar metabolism in Brevibacterium flavum. Agric. Biol. Chem. 51: 2671-2678.   DOI
8 Netzer R, Krause M, Rittmann D, Peters-Wendisch PG, Eggeling L, Wendisch VF, Sahm H. 2004. Roles of pyruvate kinase and malic enzyme in Corynebacterium glutamicum for growth on carbon sources requiring gluconeogenesis. Arch. Microbiol. 182: 354-363.   DOI
9 Parche S, Burkovski A, Sprenger GA, Weil B, Kramer R, Titgemeyer F. 2001. Corynebacterium glutamicum: a dissection of the PTS. J. Mol. Microbiol. Biotechnol. 3: 423-428.
10 Park SD, Lee SN, Park IH, Choi JS, Jeong WK, Kim YH, Lee HS. 2004. Isolation and characterization of transcriptional elements from Corynebacterium glutamicum. J. Microbiol. Biotechnol. 14: 789-795.
11 Cha PH, P ark SY, Moon MW, Subhadra B, Oh TK, K im E , Kim JF, Lee JK. 2010. Characterization of an adenylate cyclase gene (cyaB) deletion mutant of Corynebacterium glutamicum ATCC 13032. Appl. Microbiol. Biotechnol. 85: 1061-1068.   DOI   ScienceOn
12 Claes WA, Puhler 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.   DOI
13 Cocaign M, Monnet C, Lindley ND. 1993. Batch kinetics of Corynebacterium glutamicum during growth on various carbon substrates: use of substrate mixtures to localize metabolic bottlenecks. Appl. Microbiol. Biotechnol. 40: 526-530.
14 Dominguez H, Cocaign-Bousquet M, Lindley ND. 1993. Simultaneous consumption of glucose and fructose from sugar mixtures during batch growth of Corynebacterium glutamicum. Appl. Microbiol. Biotechnol. 47: 600-603.
15 Eggeling L, Bott M. 2005. Handbook of Corynebacterium glutamicum. CRC Press, Boca Raton, USA.
16 Eggeling L and Reyes O. 2004. Experiments, pp. 535-566. In Eggeling L, Bott M (ed.), Handbook of Corynebacterium glutamicum, Taylor & Francis, Boca Raton, FL.
17 Engels V, Wendisch VF. 2007. The DeoR-type regulator SugR represses expression of ptsG in Corynebacterium glutamicum. J. Bacteriol. 189: 2955-2966.   DOI   ScienceOn
18 Frunzke J, Engels V, Hasenbein S, Gatgens C, Bott M. 2008. 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.
19 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.   DOI   ScienceOn
20 Hanahan D. 1983. Studies on transformation of Escherichia coli with plasmids. J. Mol. Biol. 166: 557-580.   DOI
21 Jungwirth B, Emer D, Brune I, Hansmeier N, Puhler A, Eikmanns BJ, Tauch A. 2008. Triple transcriptional control of the resuscitation promoting factor 2 (rpf2) gene of Corynebacterium glutamicum by the regulators of acetate metabolism RamA and RamB and the cAMP-dependent regulator GlxR. FEMS Microbiol. Lett. 281: 190-197.   DOI   ScienceOn
22 Sambrook J, Fritsch EF, Maniatis T. 1989. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
23 Schreiner ND, Fiur D, Holátko J. 2005. E1 enzyme of the pyruvate dehydrogenase complex in Corynebacterium glutamicum: molecular analysis of the gene and phylogenetic aspects. J. Bacteriol. 187: 6005-6018.   DOI   ScienceOn
24 Shaw WV. 1975. Chloramphenicol acetyltransferase from chloramphenicol-resistant bacteria. Methods Enzymol. 43: 737-755.   DOI
25 Stansen C, Uy D, Delaunay S, Eggeling L, Goergen JL, Wendisch VF. 2005. Characterization of a Corynebacterium glutamicum lactate utilization operon induced during temperaturetriggered glutamate production. Appl. Environ. Microbiol. 71: 5920-5928.   DOI   ScienceOn
26 Tanaka Y, Teramoto H, Inui M, Yukawa H. 2008. Regulation of expression of general components of the phosphoenolpyruvate: carbohydrate phosphotransferase system (PTS) by the global regulator SugR in Corynebacterium glutamicum. Appl. Microbiol. Biotechnol. 78: 309-318.   DOI
27 Tauch A, Kirchner O, Loffler B, Gotker S, Puhler A, Kalinowski J. 2002. Efficient electrotransformation of Corynebacterium diphtheriae with a mini-replicon derived from the Corynebacterium glutamicum plasmid pGA1. Curr. Microbiol. 45: 362-367.   DOI   ScienceOn
28 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.   DOI   ScienceOn
29 Arndt A, Auchter M, Ishige T, Wendisch VF, Eikmanns BJ. 2008. Ethanol catabolism in Corynebacterium glutamicum. J. Mol. Microbiol. Biotechnol. 15: 222-233.   DOI   ScienceOn
30 Arndt A, Eikmanns BJ. 2007. The alcohol dehydrogenase gene adhA in Corynebacterium glutamicum is subject to carbon catabolite repression. J. Bacteriol. 189: 7408-7416.   DOI   ScienceOn
31 Auchter M, Arndt A, Eikmanns BJ. 2009. Dual transcriptional control of the acetaldehyde dehydrogenase gene ald of Corynebacterium glutamicum by RamA and RamB. J. Biotechnol. 140: 84-91.   DOI   ScienceOn
32 Bernt E, Gutmann I. 1974. Aethanol, pp. 1545-1548. In Bergmeyer HU (ed.), Methoden der Enzymatischen Analyse. Verlag Chemie, Weinheim.
33 Park SY, Moon MW, Subhadra B, Lee JK. 2010. Functional characterization of the glxR deletion mutant of Corynebacterium glutamicum ATCC 13032: involvement of GlxR in acetate metabolism and carbon catabolite repression. FEMS Microbiol. Lett. 304: 107-115.   DOI   ScienceOn