Abstract
Corynebacterium glutamicum is an important industrial producer of various amino acids with great potential for the production of other metabolites. The complete genome sequences of two C. glutamicum strains were determined and the use of genome-based approaches (transcriptomics, proteomics, metabolomics, and fluxomics) provided large amounts of data on the metabolism of this bacterium and its regulation. Many tools for genetic manipulations in C. glutamicum have been developed and used for the analysis of gene functions as well as for the construction and improvement of production strains. These tools include various types of plasmid vectors (cloning, promoter–probe, and expression vectors), DNA transfer methods, cloning heterologous genes, introducing protein secretion systems and gene replacement and genome rearrangement methods. Here we summarize the latest developments in the field of genetic engineering in C. glutamicum, give examples of the use of these new tools, and mention the challenges which stand in the way of fully implementing these tools and this acquired knowledge for the construction of superior production strains.
Similar content being viewed by others
References
Adham SA, Campelo AB, Ramos A, Gil JA (2001) Construction of a xylanase-producing strain of Brevibacterium lactofermentum by stable integration of an engineered xysA gene from Streptomyces halstedii JM8. Appl Environ Microbiol 67:5425–5430
Amador E, Martín JF, Castro JM (2000) A Brevibacterium lactofermentum 16S rRNA gene used as target site for homologous recombination. FEMS Microbiol Lett 185:199–204
Arndt A, Eikmanns BJ (2007) The alcohol dehydrogenase gene adhA in Corynebacterium glutamicum is subject to carbon catabolite repression. J Bacteriol 189:7408–7416
Asakura Y, Kimura E, Usuda Y, Kawahara Y, Matsui K, Osumi T, Nakamatsu T (2007) Altered metabolic flux due to deletion of odhA causes L-glutamate overproduction in Corynebacterium glutamicum. Appl Environ Microbiol 73:1308–1319
Barrett E, Stanton C, Zelder O, Fitzgerald G, Ross RP (2004) Heterologous expression of lactose- and galactose-utilizing pathways from lactic acid bacteria in Corynebacterium glutamicum for production of lysine in whey. Appl Environ Microbiol 70:2861–2866
Barriuso-Iglesias M, Barreiro C, Flechoso F, Martín JF (2006) Transcriptional analysis of the F0F1 ATPase operon of Corynebacterium glutamicum ATCC 13032 reveals strong induction by alkaline pH. Microbiology 152:11–21
Barriuso-Iglesias M, Schluesener D, Barreiro C, Poetsch A, Martín JF (2008) Response of the cytoplasmic and membrane proteome of Corynebacterium glutamicum ATCC 13032 to pH changes. BMC Microbiol 8:225
Bartek T, Zönnchen E, Klein B, Gerstmeir R, Makus P, Lang S, Oldiges M (2010) Analysing overexpression of L-valine biosynthesis genes in pyruvate-dehydrogenase deficient Corynebacterium glutamicum. J Ind Microbiol Biotechnol 37:263–270
Baumbach J, Wittkop T, Kleindt CK, Tauch A (2009) Integrated analysis and reconstruction of microbial transcriptional gene regulatory networks using CoryneRegNet. Nat Protoc 4:992–1005
Becker J, Klopprogge C, Herold A, Zelder O, Bolten CJ, Wittmann C (2007) Metabolic flux engineering of L-lysine production in Corynebacterium glutamicum—over expression and modification of G6P dehydrogenase. J Biotechnol 132:99–109
Becker J, Zelder O, Häfner S, Schröder H, Wittmann C (2011) From zero to hero—design-based systems metabolic engineering of Corynebacterium glutamicum for l-lysine production. Metab Eng 13:159–168
Ben-Samoun K, Leblon G, Reyes O (1999) Positively regulated expression of the Escherichia coli araBAD promoter in Corynebacterium glutamicum. FEMS Microbiol Lett 174:125–130
Brabetz W, Liebl W, Schleifer KH (1991) Studies on the utilization of lactose by Corynebacterium glutamicum, bearing the lactose operon of Escherichia coli. Arch Microbiol 155:607–612
Brune I, Brinkrolf K, Kalinowski J, Pühler A, Tauch A (2005) The individual and common repertoire of DNA-binding transcriptional regulators of Corynebacterium glutamicum, Corynebacterium efficiens, Corynebacterium diphtheriae and Corynebacterium jeikeium deduced from the complete genome sequences. BMC Genomics 6:86
Buschke N, Schröder H, Wittmann C (2011) Metabolic engineering of Corynebacterium glutamicum for production of 1,5-diaminopentane from hemicellulose. Biotechnol J 6:306–317
Carpinelli J, Krämer R, Agosin E (2006) Metabolic engineering of Corynebacterium glutamicum for trehalose overproduction: role of the TreYZ trehalose biosynthetic pathway. Appl Environ Microbiol 72:1949–1955
Chinen A, Kozlov YI, Hara Y, Izui H, Yasueda H (2007) Innovative metabolic pathway design for efficient L-glutamate production by suppressing CO2 emission. J Biosci Bioeng 103:262–269
Date M, Itaya H, Matsui H, Kikuchi Y (2006) Secretion of human epidermal growth factor by Corynebacterium glutamicum. Lett Appl Microbiol 42:66–70
Eggeling L, Reyes O (2005) Experiments. In: Eggeling L, Bott M (eds) Handbook of Corynebacterium glutamicum. CRC, Boca Raton, pp 535–566
Ehira S, Teramoto H, Inui M, Yukawa H (2009) Regulation of Corynebacterium glutamicum heat shock response by the extracytoplasmic-function sigma factor SigH and transcriptional regulators HspR and HrcA. J Bacteriol 191:2964–2972
Eikmanns BJ, Thum-Schmitz N, Eggeling L, Ludtke KU, Sahm H (1994) Nucleotide sequence, expression and transcriptional analysis of the Corynebacterium glutamicum gltA gene encoding citrate synthase. Microbiology 140:1817–1828
Fiuza M, Letek M, Leiba J, Villadangos AF, Vaquera J, Zanella-Cleon I, Mateos LM, Molle V, Gil JA (2010) Phosphorylation of a novel cytoskeletal protein (RsmP) regulates rod-shaped morphology in Corynebacterium glutamicum. J Biol Chem 285:29387–29397
Fränzel B, Poetsch A, Trötschel C, Persicke M, Kalinowski J, Wolters DA (2010a) Quantitative proteomic overview on the Corynebacterium glutamicum L-lysine producing strain DM1730. J Proteomics 73:2336–2353
Fränzel B, Trötschel C, Rückert C, Kalinowski J, Poetsch A, Wolters DA (2010b) Adaptation of Corynebacterium glutamicum to salt-stress conditions. Proteomics 10:445–457
Frunzke J, Bramkamp M, Schweitzer JE, Bott M (2008) Population heterogeneity in Corynebacterium glutamicum ATCC 13032 caused by prophage CGP3. J Bacteriol 190:5111–5119
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
Glanemann C, Loos A, Gorret N, Willis L, O’Brien XM, Lessard P, Sinskey AJ (2003) Disparity between changes in mRNA abundance and enzyme activity in Corynebacterium glutamicum: implications for DNA microarray analysis. Appl Microbiol Biotechnol 61:61–68
Hänssler E, Müller T, Palumbo K, Pátek M, Brocker M, Krämer R, Burkovski A (2009) A game with many players: control of gdh transcription in Corynebacterium glutamicum. J Biotechnol 142:114–122
Haussmann U, Qi SW, Wolters D, Rogner M, Liu SJ, Poetsch A (2009) Physiological adaptation of Corynebacterium glutamicum to benzoate as alternative carbon source—a membrane proteome-centric view. Proteomics 9:3635–3651
Hayashi M, Mizoguchi H, Shiraishi N, Obayashi M, Nakagawa S, Imai J, Watanabe S, Ota T, Ikeda M (2002) Transcriptome analysis of acetate metabolism in Corynebacterium glutamicum using a newly developed metabolic array. Biosci Biotechnol Biochem 66:1337–1344
Hayashi M, Ohnishi J, Mitsuhashi S, Yonetani Y, Hashimoto S, Ikeda M (2006) Transcriptome analysis reveals global expression changes in an industrial L-lysine producer of Corynebacterium glutamicum. Biosci Biotechnol Biochem 70:546–550
Hermann T (2003) Industrial production of amino acids by coryneform bacteria. J Biotechnol 104:155–172
Hermann T, Pfefferle W, Baumann C, Busker E, Schaffer S, Bott M, Sahm H, Dusch N, Kalinowski J, Pühler A, Bendt AK, Krämer R, Burkovski A (2001) Proteome analysis of Corynebacterium glutamicum. Electrophoresis 22:1712–1723
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
Holátko J, Elišáková V, Prouza M, Sobotka M, Nešvera J, Pátek M (2009) Metabolic engineering of the L-valine biosynthesis pathway in Corynebacterium glutamicum using promoter activity modulation. J Biotechnol 139:203–210
Hüser AT, Chassagnole C, Lindley ND, Merkamm M, Guyonvarch A, Elišáková V, Pátek M, Kalinowski J, Brune I, Pühler A, Tauch A (2005) Rational design of a Corynebacterium glutamicum pantothenate production strain and its characterization by metabolic flux analysis and genome-wide transcriptional profiling. Appl Environ Microbiol 71:3255–3268
Ikeda M, Nakagawa S (2003) The Corynebacterium glutamicum genome: features and impacts on biotechnological processes. Appl Microbiol Biotechnol 62:99–109
Inui M, Kawaguchi H, Murakami S, Vertes AA, Yukawa H (2004) Metabolic engineering of Corynebacterium glutamicum for fuel ethanol production under oxygen-deprivation conditions. J Mol Microbiol Biotechnol 8:243–254
Inui M, Suda M, Okino S, Nonaka H, Puskas LG, Vertes AA, Yukawa H (2007) Transcriptional profiling of Corynebacterium glutamicum metabolism during organic acid production under oxygen deprivation conditions. Microbiology 153:2491–2504
Ishige T, Krause M, Bott M, Wendisch VF, Sahm H (2003) The phosphate starvation stimulon of Corynebacterium glutamicum determined by DNA microarray analyses. J Bacteriol 185:4519–4529
Jäger W, Schäfer A, Pühler A, Labes G, Wohlleben W (1992) Expression of the Bacillus subtilis sacB gene leads to sucrose sensitivity in the gram-positive bacterium Corynebacterium glutamicum but not in Streptomyces lividans. J Bacteriol 174:5462–5465
Jäger W, Schäfer A, Kalinowski J, Pühler A (1995) Isolation of insertion elements from gram-positive Brevibacterium, Corynebacterium and Rhodooccus strains using the Bacillus subtilis sacB gene as a positive selection marker. FEMS Microbiol Lett 126:1–6
Jakoby M, Ngouoto-Nkili CE, Burkovski A (1999) Construction and application of new Corynebacterium glutamicum vectors. Biotechnol Tech 13:437–441
Jo SJ, Maeda M, Ooi T, Taguchi S (2006) Production system for biodegradable polyester polyhydroxybutyrate by Corynebacterium glutamicum. J Biosci Bioeng 102:233–236
Jojima T, Fujii M, Mori E, Inui M, Yukawa H (2010) Engineering of sugar metabolism of Corynebacterium glutamicum for production of amino acid L-alanine under oxygen deprivation. Appl Microbiol Biotechnol 87:159–165
Jungwirth B, Emer D, Brune I, Hansmeier N, Pühler 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
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, Wiegräbe 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
Kawaguchi H, Vertes AA, Okino S, Inui M, Yukawa H (2006) Engineering of a xylose metabolic pathway in Corynebacterium glutamicum. Appl Environ Microbiol 72:3418–3428
Kawaguchi H, Sasaki M, Vertes AA, Inui M, Yukawa H (2008) Engineering of an L-arabinose metabolic pathway in Corynebacterium glutamicum. Appl Microbiol Biotechnol 77:1053–1062
Kikuchi Y, Date M, Yokoyama K, Umezawa Y, Matsui H (2003) Secretion of active-form Streptoverticillium mobaraense transglutaminase by Corynebacterium glutamicum: processing of the pro-transglutaminase by a cosecreted subtilisin-like protease from Streptomyces albogriseolus. Appl Environ Microbiol 69:358–366
Kikuchi Y, Itaya H, Date M, Matsui K, Wu LF (2008) Production of Chryseobacterium proteolyticum protein-glutaminase using the twin-arginine translocation pathway in Corynebacterium glutamicum. Appl Microbiol Biotechnol 78:67–74
Kikuchi Y, Itaya H, Date M, Matsui K, Wu LF (2009) TatABC overexpression improves Corynebacterium glutamicum Tat-dependent protein secretion. Appl Environ Microbiol 75:603–607
Kim IK, Jeong WK, Lim SH, Hwang IK, Kim YH (2010) The small ribosomal protein S12P gene rpsL as an efficient positive selection marker in allelic exchange mutation systems for Corynebacterium glutamicum. J Microbiol Methods 84:128–130
Kind S, Jeong WK, Schröder H, Wittmann C (2010) Systems-wide metabolic pathway engineering in Corynebacterium glutamicum for bio-based production of diaminopentane. Metab Eng 12:341–351
Kirchner O, Tauch A (2003) Tools for genetic engineering in the amino acid-producing bacterium Corynebacterium glutamicum. J Biotechnol 104:287–299
Kjeldsen KR, Nielsen J (2009) In silico genome-scale reconstruction and validation of the Corynebacterium glutamicum metabolic network. Biotechnol Bioeng 102:583–597
Knoppová M, Phensaijai M, Veselý M, Zemanová M, Nešvera J, Pátek M (2007) Plasmid vectors for testing in vivo promoter activities in Corynebacterium glutamicum and Rhodococcus erythropolis. Curr Microbiol 55:234–239
Krause FS, Henrich A, Blombach B, Krämer R, Eikmanns BJ, Seibold GM (2010) Increased glucose utilization in Corynebacterium glutamicum by use of maltose, and its application for the improvement of L-valine productivity. Appl Environ Microbiol 76:370–374
Larisch C, Nakunst D, Hüser AT, Tauch A, Kalinowski J (2007) The alternative sigma factor SigB of Corynebacterium glutamicum modulates global gene expression during transition from exponential growth to stationary phase. BMC Genomics 8:4
Letek M, Valbuena N, Ramos A, Ordonez E, Gil JA, Mateos LM (2006) Characterization and use of catabolite-repressed promoters from gluconate genes in Corynebacterium glutamicum. J Bacteriol 188:409–423
Li L, Wada M, Yokota A (2007) Cytoplasmic proteome reference map for a glutamic acid-producing Corynebacterium glutamicum ATCC 14067. Proteomics 7:4317–4322
Liebl W, Sinskey AJ, Schleifer KH (1992) Expression, secretion, and processing of staphylococcal nuclease by Corynebacterium glutamicum. J Bacteriol 174:1854–1861
Liu Q, Ouyang SP, Kim J, Chen GQ (2007) The impact of PHB accumulation on L-glutamate production by recombinant Corynebacterium glutamicum. J Biotechnol 132:273–279
Liu Q, Zhang J, Wei XX, Ouyang SP, Wu Q, Chen GQ (2008) Microbial production of L-glutamate and L-glutamine by recombinant Corynebacterium glutamicum harboring Vitreoscilla hemoglobin gene vgb. Appl Microbiol Biotechnol 77:1297–1304
Loos A, Glanemann C, Willis LB, O’Brien XM, Lessard PA, Gerstmeir R, Guillouet S, Sinskey AJ (2001) Development and validation of Corynebacterium DNA microarrays. Appl Environ Microbiol 67:2310–2318
Mimitsuka T, Sawai H, Hatsu M, Yamada K (2007) Metabolic engineering of Corynebacterium glutamicum for cadaverine fermentation. Biosci Biotechnol Biochem 71:2130–2135
Moreau S, Blanco C, Trautwetter A (1999) Site-specific integration of corynephage phi16: construction of an integration vector. Microbiology 145:539–548
Muffler A, Bettermann S, Haushalter M, Horlein A, Neveling U, Schramm M, Sorgenfrei O (2002) Genome-wide transcription profiling of Corynebacterium glutamicum after heat shock and during growth on acetate and glucose. J Biotechnol 98:255–268
Nakamura J, Kanno S, Kimura E, Matsui K, Nakamatsu T, Wachi M (2006) Temperature-sensitive cloning vector for Corynebacterium glutamicum. Plasmid 56:179–186
Nakata K, Inui M, Kos PB, Vertes AA, Yukawa H (2004) Vectors for genetic engineering of corynebacteria. In: Saha BC (ed) Fermentation biotechnology. American Chemical Society, Washington, pp 175–191
Nakunst D, Larisch C, Hüser AT, Tauch A, Pühler A, Kalinowski J (2007) The extracytoplasmic function-type sigma factor SigM of Corynebacterium glutamicum ATCC 13032 is involved in transcription of disulfide stress-related genes. J Bacteriol 189:4696–4707
Nešvera J, Pátek M (2008) Plasmids and promoters in corynebacteria and their applications. In: Burkovski A (ed) Corynebacteria. Genomics and molecular biology. Caister, Norfolk, pp 113–154
Neuner A, Heinzle E (2011) Mixed glucose and lactate uptake by Corynebacterium glutamicum through metabolic engineering. Biotechnol J 6:318–329
Ogino H, Teramoto H, Inui M, Yukawa H (2008) DivS, a novel SOS-inducible cell-division suppressor in Corynebacterium glutamicum. Mol Microbiol 67:597–608
Ohnishi J, Mitsuhashi S, Hayashi M, Ando S, Yokoi H, Ochiai K, Ikeda M (2002) A novel methodology employing Corynebacterium glutamicum genome information to generate a new L-lysine-producing mutant. Appl Microbiol Biotechnol 58:217–223
Okibe N, Suzuki N, Inui M, Yukawa H (2009) Isolation, evaluation and use of two strong, carbon source-inducible promoters from Corynebacterium glutamicum. Lett Appl Microbiol 50:173–180
Okibe N, Suzuki N, Inui M, Yukawa H (2011) Efficient markerless gene replacement in Corynebacterium glutamicum using a new temperature-sensitive plasmid. J Microbiol Meth. doi:https://doi.org/10.1016/j.mimet.2011.02.012
Oram M, Woolston JE, Jacobson AD, Holmes RK, Oram DM (2007) Bacteriophage-based vectors for site-specific insertion of DNA in the chromosome of corynebacteria. Gene 391:53–62
Ozaki A, Katsumata R, Oka T, Furuya A (1984) Functional expression of the genes of Escherichia coli in gram-positive Corynebacterium glutamicum. Mol Gen Genet 196:175–178
Park JU, Jo JH, Kim YJ, Chung SS, Lee JH, Lee HH (2008) Construction of heat-inducible expression vector of Corynebacterium glutamicum and C. ammoniagenes: fusion of lambda operator with promoters isolated from C. ammoniagenes. J Microbiol Biotechnol 18:639–647
Pátek M, Nešvera J (2011a) Promoters and plasmid vectors of Corynebacterium glutamicum. In: Yukawa H, Inui M (eds) Biology and biotechnology of Corynebacterium glutamicum. Springer (in press)
Pátek M, Nešvera J (2011b) Sigma factors and promoters in Corynebacterium glutamicum. J Biotechnol. doi:https://doi.org/10.1016/j.jbiotec.2011.01.017
Pátek M, Eikmanns BJ, Pátek J, Sahm H (1996) Promoters from Corynebacterium glutamicum: cloning, molecular analysis and search for a consensus motif. Microbiology 142:1297–1309
Pátek M, Nešvera J, Guyonvarch A, Reyes O, Leblon G (2003) Promoters of Corynebacterium glutamicum. J Biotechnol 104:311–323
Peters-Wendisch PG, Schiel B, Wendisch VF, Katsoulidis E, Möckel B, Sahm H, Eikmanns BJ (2001) Pyruvate carboxylase is a major bottleneck for glutamate and lysine production by Corynebacterium glutamicum. J Mol Microbiol Biotechnol 3:295–300
Pfefferle W, Möckel B, Bathe B, Marx A (2003) Biotechnological manufacture of lysine. Adv Biochem Eng Biotechnol 79:59–112
Radford AJ, Hodgson AL (1991) Construction and characterization of a Mycobacterium–Escherichia coli shuttle vector. Plasmid 25:149–153
Ramos A, Honrubia MP, Valbuena N, Vaquera J, Mateos LM, Gil JA (2003) Involvement of DivIVA in the morphology of the rod-shaped actinomycete Brevibacterium lactofermentum. Microbiology 149:3531–3542
Rittmann D, Lindner SN, Wendisch VF (2008) Engineering of a glycerol utilization pathway for amino acid production by Corynebacterium glutamicum. Appl Environ Microbiol 74:6216–6222
Salim K, Haedens V, Content J, Leblon G, Huygen K (1997) Heterologous expression of the Mycobacterium tuberculosis gene encoding antigen 85A in Corynebacterium glutamicum. Appl Environ Microbiol 63:4392–4400
Santamaría R, Gil JA, Mesas JM, Martín JF (1984) Characterization of endogenous plasmid and development of cloning vectors and a transformation system in Brevibacterium lactofementum. J Gen Microbiol 130:2237–2246
Sato H, Orishimo K, Shirai T, Hirasawa T, Nagahisa K, Shimizu H, Wachi M (2008) Distinct roles of two anaplerotic pathways in glutamate production induced by biotin limitation in Corynebacterium glutamicum. J Biosci Bioeng 106:51–58
Schäfer A, Tauch A, Jäger W, Kalinowski J, Thierbach G, Pühler A (1994) Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum. Gene 145:69–73
Schneider J, Wendisch VF (2010) Putrescine production by engineered Corynebacterium glutamicum. Appl Microbiol Biotechnol 88:859–868
Schneider J, Niermann K, Wendisch VF (2011) Production of the amino acids L-glutamate, L-lysine, L-ornithine and L-arginine from arabinose by recombinant Corynebacterium glutamicum. J Biotechnol. doi:https://doi.org/10.1016/j.jbiotec.2010.07.009
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
Schröder J, Jochmann N, Rodionov DA, Tauch A (2010) The Zur regulon of Corynebacterium glutamicum ATCC 13032. BMC Genomics 11:12
Schweitzer JE, Stolz M, Diesveld R, Etterich H, Eggeling L (2009) The serine hydroxymethyltransferase gene glyA in Corynebacterium glutamicum is controlled by GlyR. J Biotechnol 139:214–221
Seibold G, Auchter M, Berens S, Kalinowski J, Eikmanns BJ (2006) Utilization of soluble starch by a recombinant Corynebacterium glutamicum strain: growth and lysine production. J Biotechnol 124:381–391
Silberbach M, Burkovski A (2006) Application of global analysis techniques to Corynebacterium glutamicum: new insights into nitrogen regulation. J Biotechnol 126:101–110
Smith KM, Cho KM, Liao JC (2010) Engineering Corynebacterium glutamicum for isobutanol production. Appl Microbiol Biotechnol 87:1045–1055
Sonnen H, Thierbach G, Kautz S, Kalinowski J, Schneider J, Pühler A, Kutzner HJ (1991) Characterization of pGA1, a new plasmid from Corynebacterium glutamicum LP-6. Gene 107:69–74
Srivastava P, Deb JK (2002) Construction of fusion vectors of corynebacteria: expression of glutathione-S-transferase fusion protein in Corynebacterium acetoacidophilum ATCC 21476. FEMS Microbiol Lett 212:209–216
Stäbler N, Oikawa T, Bott M, Eggeling L (2011) Corynebacterium glutamicum as a host for the synthesis and export of D-amino acids. J Bacteriol. doi:https://doi.org/10.1128/JB.01295-10
Suzuki N, Nonaka H, Tsuge Y, Inui M, Yukawa H (2005a) New multiple-deletion method for the Corynebacterium glutamicum genome, using a mutant lox sequence. Appl Environ Microbiol 71:8472–8480
Suzuki N, Nonaka H, Tsuge Y, Okayama S, Inui M, Yukawa H (2005b) Multiple large segment deletion method for Corynebacterium glutamicum. Appl Microbiol Biotechnol 69:151–161
Suzuki N, Tsuge Y, Inui M, Yukawa H (2005c) Cre/loxP-mediated deletion system for large genome rearrangements in Corynebacterium glutamicum. Appl Microbiol Biotechnol 67:225–233
Suzuki N, Okai N, Nonaka H, Tsuge Y, Inui M, Yukawa H (2006) High-throughput transposon mutagenesis of Corynebacterium glutamicum and construction of a single-gene disruptant mutant library. Appl Environ Microbiol 72:3750–3755
Suzuki N, Watanabe K, Okibe N, Tsuchida Y, Inui M, Yukawa H (2009) Identification of new secreted proteins and secretion of heterologous amylase by C. glutamicum. Appl Microbiol Biotechnol 82:491–500
Tateno T, Fukuda H, Kondo A (2007a) Direct production of L-lysine from raw corn starch by Corynebacterium glutamicum secreting Streptococcus bovis alpha-amylase using cspB promoter and signal sequence. Appl Microbiol Biotechnol 77:533–541
Tateno T, Fukuda H, Kondo A (2007b) Production of L-lysine from starch by Corynebacterium glutamicum displaying alpha-amylase on its cell surface. Appl Microbiol Biotechnol 74:1213–1220
Tateno T, Hatada K, Tanaka T, Fukuda H, Kondo A (2009a) Development of novel cell surface display in Corynebacterium glutamicum using porin. Appl Microbiol Biotechnol 84:733–739
Tateno T, Okada Y, Tsuchidate T, Tanaka T, Fukuda H, Kondo A (2009b) Direct production of cadaverine from soluble starch using Corynebacterium glutamicum coexpressing alpha-amylase and lysine decarboxylase. Appl Microbiol Biotechnol 82:115–121
Tauch A (2005) Native plasmids of amino acid-producing corynebacteria. In: Eggeling L, Bott M (eds) Handbook of Corynebacterium glutamicum. CRC, Boca Raton, pp 57–80
Tauch A, Götker S, Pühler A, Kalinowski J, Thierbach G (2002) The alanine racemase gene alr is an alternative to antibiotic resistance genes in cloning systems for industrial Corynebacterium glutamicum strains. J Biotechnol 99:79–91
Tauch A, Pühler A, Kalinowski J, Thierbach G (2003) Plasmids in Corynebacterium glutamicum and their molecular classification by comparative genomics. J Biotechnol 104:27–40
Tsuchidate T, Tateno T, Okai N, Tanaka T, Ogino C, Kondo A (2011) Glutamate production from β-glucan using endoglucanase-secreting Corynebacterium glutamicum. Appl Microbiol Biotechnol. doi:https://doi.org/10.1007/s00253-011-3116-7
Tsuchiya M, Morinaga Y (1988) Genetic control systems of Escherichia coli can confer inducible expression of cloned genes in coryneform bacteria. Bio/Technology 6:428–430
Tsuge Y, Suzuki N, Inui M, Yukawa H (2007a) Random segment deletion based on IS31831 and Cre/loxP excision system in Corynebacterium glutamicum. Appl Microbiol Biotechnol 74:1333–1341
Tsuge Y, Suzuki N, Ninomiya K, Inui M, Yukawa H (2007b) Isolation of a new insertion sequence, IS13655, and its application to Corynebacterium glutamicum genome mutagenesis. Biosci Biotechnol Biochem 71:1683–1690
Vašicová P, Abrhámová Z, Nešvera J, Pátek M, Sahm H, Eikmanns B (1998) Integrating and autonomously replicating vectors for analysis of promoters in Corynebacterium glutamicum. Biotechnol Tech 12:743–746
Vašicová P, Pátek M, Nešvera J, Sahm H, Eikmanns B (1999) Analysis of the Corynebacterium glutamicum dapA promoter. J Bacteriol 181:6188–6191
Vertes AA, Inui M, Kobayashi M, Kurusu Y, Yukawa H (1994) Isolation and characterization of IS31831, a transposable element from Corynebacterium glutamicum. Mol Microbiol 11:739–746
Wendisch VF (2003) Genome-wide expression analysis in Corynebacterium glutamicum using DNA microarrays. J Biotechnol 104:273–285
Wendisch VF, Bott M, Kalinowski J, Oldiges M, Wiechert W (2006) Emerging Corynebacterium glutamicum systems biology. J Biotechnol 124:74–92
Wittmann C, Heinzle E (2002) Genealogy profiling through strain improvement by using metabolic network analysis: metabolic flux genealogy of several generations of lysine-producing corynebacteria. Appl Environ Microbiol 68:5843–5859
Woo HM, Noack S, Seibold GM, Willbold S, Eikmanns BJ, Bott M (2010) Link between phosphate starvation and glycogen metabolism in Corynebacterium glutamicum, revealed by metabolomics. Appl Environ Microbiol 76:6910–6919
Xu D, Tan Y, Huan X, Hu X, Wang X (2010) Construction of a novel shuttle vector for use in Brevibacterium flavum, an industrial amino acid producer. J Microbiol Methods 80:86–92
Yao W, Deng X, Liu M, Zheng P, Sun Z, Zhang Y (2009) Expression and localization of the Corynebacterium glutamicum NCgl1221 protein encoding an L-glutamic acid exporter. Microbiol Res 164:680–687
Yasuda K, Jojima T, Suda M, Okino S, Inui M, Yukawa H (2007) Analyses of the acetate-producing pathways in Corynebacterium glutamicum under oxygen-deprived conditions. Appl Microbiol Biotechnol 77:853–860
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
Yukawa H, Omumasaba CA, Nonaka H, Kos P, Okai N, Suzuki N, Suda M, Tsuge Y, Watanabe J, Ikeda Y, Vertes AA, Inui M (2007) Comparative analysis of the Corynebacterium glutamicum group and complete genome sequence of strain R. Microbiology 153:1042–1058
Acknowledgments
This work was supported by Grant 204/09/J015 from the Scientific Council of the Czech Republic and by Institutional Research Concept No. AV0Z50200510.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Nešvera, J., Pátek, M. Tools for genetic manipulations in Corynebacterium glutamicum and their applications. Appl Microbiol Biotechnol 90, 1641–1654 (2011). https://doi.org/10.1007/s00253-011-3272-9
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-011-3272-9