Abstract
l-lysine is made in an exceptional large quantity of currently 2,200,000 tons/year and belongs therefore to one of the leading biotechnological products. Production is done almost exclusively with mutants of Corynebacterium glutamicum. The increasing l-lysine market forces companies to improve the production process fostering also a deeper understanding of the microbial physiology of C. glutamicum. Current major challenges are the identification of ancillary mutations not intuitively related with product increase. This review gives insights on how cellular characteristics enable to push the carbon flux in metabolism towards its theoretical maximum, and this example may also serve as a guide to achieve and increase the formation of other products of interest in microbial biotechnology.
Similar content being viewed by others
References
Bankar SB, Singhal RS (2010) Optimization of poly-epsilon-lysine production by Streptomyces noursei NRRL 5126. Bioresour Technol 101:8370–8375
Bartek T, Blombach B, Lang S, Eikmanns BJ, Wiechert W, Oldiges M, Noh K, Noack S (2011) Comparative C-13 metabolic flux analysis of pyruvate dehydrogenase complex-deficient, L-valine-producing Corynebacterium glutamicum. Appl Environ Microbiol 77:6644–6652
Becker J, Klopprogge C, Zelder O, Heinzle E, Wittmann C (2005) Amplified expression of fructose 1,6-bisphosphatase in Corynebacterium glutamicum increases in vivo flux through the pentose phosphate pathway and lysine production on different carbon sources. Appl Environ Microbiol 71:8587–8596
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, Klopprogge C, Schroder H, Wittmann C (2009) Metabolic engineering of the tricarboxylic acid cycle for improved lysine production by Corynebacterium glutamicum. Appl Environ Microbiol 75:7866–7869
Becker J, Zelder O, Hafner S, Schröder H, Wittmann C (2011) From zero to hero–design-based systems metabolic engineering of Corynebacterium glutamicumfor L-lysine production. Metab Eng 13:159–168
Binder S, Schendzielorz G, Stäbler N, Krumbach K, Hoffmann K, Bott M, Eggeling L (2012) A high-throughput approach to identify genomic variants of bacterial metabolite producers at the single-cell level. Genome Biol 13:R40
Binder S, Siedler S, Marienhagen J, Bott M, Eggeling L (2013) Recombineering in Corynebacterium glutamicum combined with optical nanosensors: a general strategy for fast producer strain generation. Nucleic Acids Res 41:6360–6369
Blombach B, Hans S, Bathe B, Eikmanns BJ (2009) Acetohydroxyacid synthase, a novel target for improvement of L-lysine production by Corynebacterium glutamicum. Appl Environ Microbiol 75:419–427
Bommareddy RR, Chen Z, Rappert S, Zeng AP (2014) A de novo NADPH generation pathway for improving lysine production of Corynebacterium glutamicum by rational design of the coenzyme specificity of glyceraldehyde 3-phosphate dehydrogenase. Metab Eng 25:30–37
Buchholz J, Schwentner A, Brunnenkan B, Gabris C, Grimm S, Gerstmeir R, Takors R, Eikmanns BJ, Blombach B (2013) Platform engineering of Corynebacterium glutamicum with reduced pyruvate dehydrogenase complex activity for improved production of L-lysine, L-valine, and 2-ketoisovalerate. Appl Environ Microbiol 79:5566–5575
Buchholz J, Graf M, Freund A, Busche T, Kalinowski J, Blombach B, Takors R (2014) CO(2)/HCO(3)(−) perturbations of simulated large scale gradients in a scale-down device cause fast transcriptional responses in Corynebacterium glutamicum. Appl Microbiol Biotechnol 98:8563–8572
Burkovski A (2008) Corynebacteria, Genomics and molecular biology. Caister Academic Press, Norfolk, p 340
Buschke N, Schäfer R, Becker J, Wittmann C (2013) Metabolic engineering of industrial platform microorganisms for biorefinery applications—optimization of substrate spectrum and process robustness by rational and evolutive strategies. Bioresour Technol 135:544–554
Chen Z, Meyer W, Rappert S, Sun J, Zeng A-P (2011) Coevolutionary analysis enabled rational deregulation of allosteric enzyme inhibition in Corynebacterium glutamicum for lysine production. Appl Environ Microbiol 77:4352–4360
Chen Z, Bommareddy RR, Frank D, Rappert S, Zeng AP (2013) Deregulation of feedback inhibition of phosphoenolpyruvate carboxylase for improved lysine production in Corynebacterium glutamicum. Appl Environ Microbiol 80:1388–1393
Diesveld R, Tietze N, Fürst O, Reth A, Bathe B, Sahm H, Eggeling L (2009) Activity of exporters of Escherichia coli in Corynebacterium glutamicum, and their use to increase L-threonine production. J Mol Microbiol Biotechnol 16:198–207
Eggeling L (2009) Microbial metabolite export in biotechnology. Wiley, Hoboken
Eggeling L, Bott M (2005) Handbook of Corynebacterium glutamicum. Taylor & Francis, Boca Raton
Eggeling L, Sahm H (1999) L-Glutamate and L-lysine: traditional products with impetuous developments. Appl Microbiol Biotechnol 52:146–153
Eggeling L, Oberle S, Sahm H (1998) Improved L-lysine yield with Corynebacterium glutamicum: use of dapA resulting in increased flux combined with growth limitation. Appl Microbiol Biotechnol 49:24–30
Frommeyer M, Wiefel L, Steinbüchel A (2014) Features of the biotechnologically relevant polyamide family “cyanophycins” and their biosynthesis in prokaryotes and eukaryotes. Crit Rev Biotechnol 30:1–12
Gao BL, Gupta RS (2012) Phylogenetic framework and molecular signatures for the main clades of the phylum actinobacteria. Microbiol Mol Biol Rev 76:66–112
Georgi T, Rittmann D, Wendisch VF (2005) Lysine and glutamate production by Corynebacterium glutamicum on glucose, fructose and sucrose: roles of malic enzyme and fructose-1,6-bisphosphatase. Metab Eng 7:291–301
Hayashi M, Tabata K (2013) Metabolic engineering for L-glutamine overproduction by using DNA gyrase mutations in Escherichia coli. Appl Environ Microbiol 79:3033–3039
Hayashi M, Ohnishi J, Mitsuhashi S, Yonetani Y, Hashimoto S-I, Ikeda M (2006) Transcriptome analysis reveals global expression changes in an industrial L-lysine producer of Corynebacterium glutamicum. Biosci Biotechnol Biochem 70:546–550
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
Ikeda M, Ohnishi J, Hayashi M, Mitsuhashi S (2006) A genome-based approach to create a minimally mutated Corynebacterium glutamicum strain for efficient L-lysine production. J Ind Microbiol Biotechnol 33:610–615
Ikeda M, Mitsuhashi S, Tanaka K, Hayashi M (2009) Reengineering of a Corynebacterium glutamicum L-arginine and L-citrulline producer. Appl Environ Microbiol 75:1635–1641
Ikeda M, Mizuno Y, Awane S, Hayashi M, Mitsuhashi S, Takeno S (2011) Identification and application of a different glucose uptake system that functions as an alternative to the phosphotransferase system in Corynebacterium glutamicum. Appl Microbiol Biotechnol 90:1443–1451
Kabus A, Georgi T, Wendisch VF, Bott M (2007) Expression of the Escherichia coli pntAB genes encoding a membrane-bound transhydrogenase in Corynebacterium glutamicum improves L-lysine formation. Appl Microbiol Biotechnol 75:47–53
Käß F, Hariskos I, Michel A, Brandt HJ, Spann R, Junne S, Wiechert W, Neubauer P, Oldiges M (2014) Assessment of robustness against dissolved oxygen/substrate oscillations for C. glutamicum DM1933 in two-compartment bioreactor. Bioprocess Biosyst Eng 37:1151–1162
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
Kelle R, Hermann T, Bathe B (2005) L-lysine production. CRC Press, Taylor & Francis Group, Boca Raton
Kind S, Becker J, Wittmann C (2013) Increased lysine production by flux coupling of the tricarboxylic acid cycle and the lysine biosynthetic pathway—metabolic engineering of the availability of succinyl-CoA in Corynebacterium glutamicum. Metab Eng 15:184–195
Kind S, Neubauer S, Becker J, Yamamoto M, Völkert M, Abendroth G, Zelder O, Wittmann C (2014) From zero to hero—production of bio-based nylon from renewable resources using engineered Corynebacterium glutamicum. Metab Eng 25:113–123
Klaffl S, Brocker M, Kalinowski J, Eikmanns BJ, Bott M (2013) Complex regulation of the phosphoenolpyruvate carboxykinase gene pck and characterization of its GntR-type regulator IolR as a repressor of myo-inositol utilization genes in Corynebacterium glutamicum. J Bacteriol 195:4283–4296
Krings E, Krumbach K, Bathe B, Kelle R, Wendisch VF, Sahm H, Eggeling L (2006) Characterization of myo-inositol utilization by Corynebacterium glutamicum: the stimulon, identification of transporters, and influence on L-lysine formation. J Bacteriol 188:8054–8061
Lindner SN, Seibold GM, Henrich A, Kramer R, Wendisch VF (2011) Phosphotransferase system-independent glucose utilization in Corynebacterium glutamicum by inositol permeases and glucokinases. Appl Environ Microbiol 77:3571–3581
Marx A, Hans S, Möckel B, Bathe B, de Graaf AA, McCormack AC, Stapleton C, Burke K, O’Donohue M, Dunican LK (2003) Metabolic phenotype of phosphoglucose isomerase mutants of Corynebacterium glutamicum. J Biotechnol 104:185–197
Meiswinkel TM, Gopinath V, Lindner SN, Nampoothiri KM, Wendisch VF (2013) Accelerated pentose utilization by Corynebacterium glutamicum for accelerated production of lysine, glutamate, ornithine and putrescine. Microb Biotechnol 6:131–140
Mimitsuka T, Sawai H, Hatsu M, Yamada K (2007) Metabolic engineering of Corynebacterium glutamicum for cadaverine fermentation. Biosci Biotechnol Biochem 71:2130–2135
Mishra AK, Driessen NN, Appelmelk BJ, Besra GS (2011) Lipoarabinomannan and related glycoconjugates: structure, biogenesis and role in Mycobacterium tuberculosis physiology and host-pathogen interaction. FEMS Microbiol Rev 35:1126–1157
Mustafi N, Grünberger A, Kohlheyer D, Bott M, Frunzke J (2012) The development and application of a single-cell biosensor for the detection of L-methionine and branched-chain amino acids. Metab Eng 14:449–457
Neuner A, Wagner I, Sieker T, Ulber R, Schneider K, Peifer S, Heinzle E (2013) Production of L-lysine on different silage juices using genetically engineered Corynebacterium glutamicum. J Biotechnol 163:217–224
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
Nishio Y, Usuda Y, Matsui K, Kurata H (2008) Computer-aided rational design of the phosphotransferase system for enhanced glucose uptake in Escherichia coli. Mol Syst Biol 4:160
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
Ohnishi J, Katahira R, Mitsuhashi S, Kakita S, Ikeda M (2005) A novel gnd mutation leading to increased L-lysine production in Corynebacterium glutamicum. FEMS Microbiol Lett 242:265–274
Petersen S, Mack C, de Graaf AA, Riedel C, Eikmanns BJ, Sahm H (2001) Metabolic consequences of altered phosphoenolpyruvate carboxykinase activity in Corynebacterium glutamicum reveal anaplerotic regulation mechanisms in vivo. Metab Eng 3:344–361
Sallam A, Steinbüchel A (2010) Dipeptides in nutrition and therapy: cyanophycin-derived dipeptides as natural alternatives and their biotechnological production. Appl Microbiol Biotechnol 87:815–828
Schendzielorz G, Dippong M, Grunberger A, Kohlheyer D, Yoshida A, Binder S, Nishiyama C, Nishiyama M, Bott M, Eggeling L (2013) Taking control over control: use of product sensing in single cells to remove flux control at key enzymes in biosynthesis pathways. ACS Synth Biol 3:21–29
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
Shih IL, Shen MH, Van YT (2006) Microbial synthesis of poly(epsilon-lysine) and its various applications. Bioresour Technol 97:1148–1159
Takeno S, Murata R, Kobayashi R, Mitsuhashi S, Ikeda M (2010) Engineering of Corynebacterium glutamicum with an NADPH-generating glycolytic pathway for L-lysine production. Appl Environ Microbiol 76:7154–7160
Takors R, Bathe B, Rieping M, Hans S, Kelle R, Huthmacher K (2007) Systems biology for industrial strains and fermentation processes—example: amino acids. J Biotechnol 129:181–190
Tateno T, Fukuda H, Kondo A (2007) 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
Tsuruta H, Paddon CJ, Eng D, Lenihan JR, Horning T, Anthony LC, Regentin R, Keasling JD, Renninger NS, Newman JD (2009) High-level production of amorpha-4,11-diene, a precursor of the antimalarial agent artemisinin, in Escherichia coli. Plos One 4:e4489
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 109:2070–2081
Witthoff S, Mühlroth A, Marienhagen J, Bott M (2013) C1 metabolism in Corynebacterium glutamicum: an endogenous pathway for oxidation of methanol to carbon dioxide. Appl Environ Microbiol 79:6974–6983
Yukawa H, Inui M (2013) Corynebacterium glutamicum: biology and biotechnology. Springer, Heidelberg, New York
Acknowledgments
This work was funded by the German ministry of education and research (Grant no. 0315589A).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Eggeling, L., Bott, M. A giant market and a powerful metabolism: l-lysine provided by Corynebacterium glutamicum . Appl Microbiol Biotechnol 99, 3387–3394 (2015). https://doi.org/10.1007/s00253-015-6508-2
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-015-6508-2