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Overproduction of l-Glutamate in Corynebacterium glutamicum

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Abstract

l-Glutamic acid (l-Glu) is an industrially important amino acid for the production of the flavor enhancer, monosodium-l-glutamate (MSG), which expresses a savory taste known as umami. The fundamental fermentation process for MSG production was established in 1956 and is based on the l-glutamate-producing bacterium Corynebacterium glutamicum, which was originally isolated from a soil sample. Since the discovery of this epoch-making production method, extensive metabolic engineering studies designed to improve l-glutamate production have been conducted using the genomic information of C. glutamicum. This chapter focuses on recent studies of the molecular mechanism of l-glutamate secretion and metabolic pathway design for efficient overproduction of l-glutamate from glucose in C. glutamicum strains. The mechanism of l-glutamate overproduction is unique, and it has been demonstrated that the product of the NCgl1221 gene, which is homologous to mechano-sensitive channels, plays a crucial role in l-glutamate secretion. It has also been shown that the specific activity of the 2-oxoglutarate dehydrogenase complex decreases during l-glutamate overproduction and that the enzymatic activity is controlled by a novel regulatory mechanism that involves OdhI and serine/threonine protein kinases. On the other hand, a new metabolic pathway was designed to enhance l-glutamate production by bypassing the CO2-releasing pyruvate dehydrogenase reaction in the glycolytic pathway by introducing heterologous phosphoketolase. This attempt improved the conversion yield of l-Glu and reduced the CO2 emission level during the fermentation process of l-glutamate overproduction by C. glutamicum.

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References

  • Ajinomoto (2012) Fact sheet: food products business. http://www.ajinomoto.com/jp/ir/ebook/food-oct2012.htm

  • 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

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Börngen K, Battle AR, Möker N, Morbach S, Marin K, Martinac B, Krämer R (2010) The properties and contribution of the Corynebacterium glutamicum MscS variant to fine-tuning of osmotic adaptation. Biochim Biophys Acta 1798:2141–2149

    Article  PubMed  Google Scholar 

  • Boulahya K-A, Guedon E, Delaunay S, Schultz C, Boudrant J, Bott M, Goergen J-L (2010) OdhI dephosphorylation kinetics during different glutamate production processes involving Corynebacterium glutamicum. Appl Microbiol Biotechnol 87:1867–1874

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Goldberg ML, Racker E (1962) Formation and isolation of a glycolaldehyde-phosphoketolase intermediate. J Biol Chem 237:3841–3842

    CAS  PubMed  Google Scholar 

  • Hashimoto K, Nakamura K, Kuroda T, Yabe I, Nakamatsu T, Kawasaki H (2010) The protein encoded by NCgl1221 in Corynebacterium glutamicum functions as a mechanosensitive channel. Biosci Biotechnol Biochem 74:2546–2549

    Article  CAS  PubMed  Google Scholar 

  • Hashimoto K, Murata J, Konishi T, Yabe I, Nakamatsu T, Kawasaki H (2012) Glutamate is excreted across the cytoplasmic membrane through the NCgl1221 channel of Corynebacterium glutamicum by passive diffusion. Biosci Biotechnol Biochem 76:1422–1424

    Article  CAS  PubMed  Google Scholar 

  • Heath EC, Hurwitz J, Horecker BL, Ginsburg A (1958) Pentose fermentation by Lactobacillus plantarum. I. The cleavage of xylulose 5-phosphate by a phosphoketolase. J Biol Chem 231:1009–1029

    CAS  PubMed  Google Scholar 

  • Ikeda M, Nakagawa S (2003) The Corynebacterium glutamicum genome: features and impacts on biotechnological processes. Appl Microbiol Biotechnol 62:99–109

    Article  CAS  PubMed  Google Scholar 

  • 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, Tach A (2003) The complete Corynebacterium glutamicum ATCC13032 genome sequence and its impact on the production of L-aspartate-derived amino acids and vitamins. J Biotechnol 104:5–25

    Article  CAS  PubMed  Google Scholar 

  • Kataoka M, Hashimoto K-I, 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

    Article  CAS  PubMed  Google Scholar 

  • Kawahara Y, Takahasi-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

    Article  CAS  PubMed  Google Scholar 

  • Kim J, Hirasawa T, Saito M, Furusawa C, Shimizu H (2011) Investigation of phosphorylation status of OdhI protein during penicillin- and Tween 40-triggered glutamate overproduction by Corynebacterium glutamicum. Appl Microbiol Biotechnol 91:143–151

    Article  CAS  PubMed  Google Scholar 

  • Kimura E (2005) l-Glutamate production. In: Eggeling L, Bott M (eds) Handbook of Corynebacterium glutamicum. CRC/Taylor & Francis, Boca Raton, pp 439–464

    Google Scholar 

  • Kimura E, Abe C, Kawahara Y, Nakamatsu T, Tokuda H (1997) A dtsR1 gene-disrupted mutant of Brevibacterium lactofermentum requires fatty acids for growth and efficiently produces l-glutamate in the presence of an excess of biotin. Biochem Biophys Res Commun 234:157–161

    Google Scholar 

  • Kinoshita S, Udaka S, Shimono M (1957) Studies on the amino acid fermentation. Part I. Production of l-glutamic acid by various microorganisms. J Gen Appl Microbiol 3:193–205

    Article  CAS  Google Scholar 

  • Levina N, Totemeyer S, Stokes NR, Louis P, Jones MA, Booth IR (1999) Protection of Escherichia coli cells against extreme turgor by activation of MscS and MscL mechanosensitive channels: identification of genes required for MscS activity. EMBO J 18:1730–1737

    Article  CAS  PubMed  Google Scholar 

  • Nakamura E (2011) One hundred years since the discovery of the “umami” taste from seaweed broth by Kikunae Ikeda, who transcended his time. Chem Asian J 6:1659–1663

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Sano C (2009) History of glutamate production. Am J Clin Nutr 90:728S–732S

    Article  CAS  PubMed  Google Scholar 

  • Schluesener D, Fischer F, Kruip J, Rögner M, Poetsch A (2005) Mapping and membrane proteome of Corynebacterium glutamicum. Proteomics 5:1317–1330

    Article  CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Shimizu H, Hirasawa T (2007) Production of glutamate and glutamate-related amino acids: molecular mechanism analysis and metabolic engineering. In: Wendisch VF (ed) Amino acid biosynthesis: pathways, regulation and metabolic engineering. Springer, Berlin, pp 1–38

    Chapter  Google Scholar 

  • Shirai T, Nakato A, Izutani N, Nagahisa K, Shioya S, Kimura E, Kawarabayashi 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

    Article  CAS  PubMed  Google Scholar 

  • Strelkov S, von Elstermann M, Schomburg D (2004) Comprehensive analysis of metabolites in Corynebacterium glutamicum by gas chromatography/mass spectrometry. Biol Chem 385:853–861

    Article  CAS  PubMed  Google Scholar 

  • Suzuki R, Katayama T, Kim B-J, Wakagi T, Shoun H, Ashida H, Yamamoto K, Fushinobu S (2010) Crystal structures of phosphoketolase. Thiamine diphosphate-dependent dehydration mechanism. J Biol Chem 285:34279–34287

    Article  CAS  PubMed  Google Scholar 

  • Takahashi K, Tagami U, Shimba N, Kashiwagi T, Ishikawa K, Suzuki E (2010) Crystal structure of Bifidobacterium longum phosphoketolase; key enzyme for glucose metabolism in Bifidobacterium. FEBS Lett 584:3855–3861

    Article  CAS  PubMed  Google Scholar 

  • Wendisch VF (2003) Genome-wide expression analysis in Corynebacterium glutamicum using DNA microarrays. J Biotechnol 104:273–285

    Article  CAS  PubMed  Google Scholar 

  • Yamada K, Komagata K (1972) Taxonomic studies on coryneform bacteria. IV. Morphological, cultural, biochemical, and physiological characteristics. J Gen Appl Microbiol 18:399–416

    Article  Google Scholar 

  • Yamaguchi S, Ninomiya K (2000) Umami and food palatability. J Nutr 130(4):921S–926S

    CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Hisashi Yasueda .

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Yasueda, H. (2014). Overproduction of l-Glutamate in Corynebacterium glutamicum . In: Anazawa, H., Shimizu, S. (eds) Microbial Production. Springer, Tokyo. https://doi.org/10.1007/978-4-431-54607-8_15

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