Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Production of uridine 5′-monophosphate by Corynebacterium ammoniagenes ATCC 6872 using a statistically improved biocatalytic process

  • 323 Accesses

  • 21 Citations


Attempts were made with success to develop a two-step biocatalytic process for uridine 5′-monophosphate (UMP) production from orotic acid by Corynebacterium ammoniagenes ATCC 6872: the strain was first cultivated in a high salt mineral medium, and then cells were harvested and used as the catalyst in the UMP production reaction. Effects of cultivation and reaction conditions on UMP production were investigated. The cells exhibited the highest biocatalytic ability when cultivated in a medium containing corn steep liquor at pH 7.0 for 15 h in the exponential phase of growth. To optimize the reaction, both “one-factor-at-a-time” method and statistical method were performed. By “one-factor-at-a-time” optimization, orotic acid, glucose, phosphate ion (equimolar KH2PO4 and K2HPO4), MgCl2, Triton X-100 were shown to be the optimum components for the biocatalytic reaction. Phosphate ion and C. ammoniagenes cell were furthermore demonstrated as the most important main effects on UMP production by Plackett–Burman design, indicating that 5-phosphoribosyl-1-pyrophosphate (PRPP) synthesis was the rate-limiting step for pyrimidine nucleotides production. Optimization by a central composition design (CCD) was then performed, and up to 32 mM (10.4 g l−1) UMP was accumulated in 24 h from 38.5 mM (6 g l−1) orotic acid. The yield was threefold higher than the original UMP yield before optimization.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5


  1. Abbouni B, Elhariry HM, Auling G (2004) Overproduction of NAD+ and 5′-inosine monophosphate in the presence of 10 μM Mn2+ by a mutant of Corynebacterium ammoniagenes with thermosensitive nucleotide reduction (nrd ts) after temperature shift. Arch Microbiol 182:119–125

  2. Auling G, Thaler M, Diekmann H (1980) Parameters of unbalanced growth and reversible inhibition of deoxyribonucleic acid synthesis in Brevibacterium ammoniagenes ATCC 6872 induced by depletion of Mn2+. Inhibitor studies on the reversibility of deoxyribonucleic acid synthesis. Arch Microbiol 127:105–114

  3. Brown PR, Robb CS, Geldart SE (2002) Perspectives on analyses of nucleic acid constituents: the basis of genomics. J Chromatogr A 965:163–173

  4. Chen QH, He GQ, Ali MAM (2002) Optimization of medium composition for the production of elastase by Bacillus sp. EL31410 with response surface methodology. Enzyme Microb Technol 30:667–672

  5. Endo T, Koizumi S, Tabata K, Ozaki A (2000) Large-scale production of CMP-NeuAc and sialylated oligosaccharides through bacterial coupling. Appl Microbiol Biotechnol 53:257–261

  6. Endo T, Koizumi S, Tabata K, Kakita S, Ozaki A (2001) Large-scale production of the carbohydrate portion of the sialyl-Tn epitope, α-Neup5Ac-(2-6)-d-GalpNAc, through bacterial coupling. Carbohydr Res 330:439–443

  7. Fujio T, Maruyama A (1997) Enzymatic production of pyrimidine nucleotides using Corynebacterium ammoniagenes cells and recombinant Escherichia coli cells: enzymatic production of CDP-choline from orotic acid and choline chloride (Part I). Biosci Biotechnol Biochem 61:956–959

  8. Henriksen A, Aghajari N, Jensen KF, Gajhede M (1996) A flexible loop at the dimer interface is a part of the active site of the adjacent monomer of Escherichia coli orotate phosphoribosyltransferase. Biochemistry 35:3803–3809

  9. Kamada N, Yasuhara A, Ikeda M (2003) Significance of the non-oxidative route of the pentose phosphate pathway for supplying carbon to the purine-nucleotide pathway in Corynebacterium ammoniagenes. J Ind Microbiol Biotechnol 30:129–132

  10. Lee HC, Lee S, Sohng JK, Liou K (2004) One-pot enzymatic synthesis of UDP-d-glucose from UMP and glucose-1-phosphate using an ATP regeneration system. J Biochem Mol Biol 37:503–506

  11. Maruyama A, Fujio T (2001) ATP production from adenine by a self-coupling enzymatic process: high-level accumulation under ammonium-limited conditions. Biosci Biotechnol Biochem 65:644–650

  12. Nahálka J, Liu Z, Germeiner P, Wang P (2002) Nucleoside triphosphates production using recombinant Escherichia coli entrapped in calcium pectate gel. Biotechnol Lett 24:925–930

  13. Nakayama K, Tanaka H (1971) Production of nucleic acid-related substances by fermentative processes XXXVIII. Production of uridine 5′-monophosphate and orotidine 5′-monophosphate by Brevibacterium ammoniagenes. Agr Biol Chem 35:518–525

  14. Naveena BJ, Altaf M, Bhadriah K, Reddy G (2005) Selection of medium components by Plackett–Burman design for production of L(+) lactic acid by Lactobacillus amylophilus GV6 in SSF using wheat bran. Bioresour Technol 96:485–490

  15. Nudler AA, Garibyan AG, Bourd GI (1991) The derepression of enzymes of de novo pyrimidine biosynthesis pathway in Brevibacterium ammoniagenes producing uridine-5-monophosphate and uracil. FEMS Microbiol Lett 66:263–266

  16. Oka T, Udagawa K, Kinoshita S (1968) Unbalanced growth death due to depletion of Mn2+ in Brevibacterium ammoniagenes. J Bacteriol 96:1760–1767

  17. Plackett R, Burman J (1946) The design of optimum multifactorial experiments. Biometrika 33:305–325

  18. Stowe RA, Mayer RP (1966) Efficient screening of process variables. Ind Eng Chem 58:36–40

  19. Tabata K, Koizumi S, Endo T, Ozaki A (2000) Production of UDP-N-acetylglucosamine by coupling metabolically engineered bacteria. Biotechnol Lett 22:479–483

  20. Tortora GJ, Funke BR, Case CL (1992) Microbiology: an introduction. Benjamin/Cummings, Redwood City California

  21. Varki A (1993) Biological roles of oligosaccharides: all of the theories are correct. Glycobiology 3:97–130

Download references


The work was supported by the Scientific Research Foundation for the Returned Overseas Chinese Scholars and Key Foundation of the State Education Ministry (Grant no. 106102) and a grant from the Ministry of Science and Technology of China (National Basic Research Program of China, 2007CB707803). The authors also gratefully acknowledge the financial support of this work by Shanghai Apple Flavor & Fragrance Co., Ltd. (China).

Author information

Correspondence to Ping Xu.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Wang, X., Wang, X., Yin, M. et al. Production of uridine 5′-monophosphate by Corynebacterium ammoniagenes ATCC 6872 using a statistically improved biocatalytic process. Appl Microbiol Biotechnol 76, 321–328 (2007). https://doi.org/10.1007/s00253-007-1013-x

Download citation


  • Corynebacterium ammoniagenes
  • Uridine 5′-monophosphate
  • Biocatalytic production
  • Statistical optimization