Applied Microbiology and Biotechnology

, Volume 98, Issue 16, pp 6983–6989 | Cite as

Filamentous fungi in microtiter plates—an easy way to optimize itaconic acid production with Aspergillus terreus

  • Antje Hevekerl
  • Anja Kuenz
  • Klaus-Dieter Vorlop
Biotechnological products and process engineering


Itaconic acid is an important industrial building block and is produced by the filamentous fungi Aspergillus terreus. To make the optimization process more efficient, a scale-down from shake flasks to microtiter plates was performed. This resulted in comparable product formations, and 87.7 g/L itaconic acid was formed after 10 days of cultivation in the microtiter plate. The components of the minimal medium were varied independently for a media optimization. This resulted in an increase of the itaconic acid concentration by a variation of the KH2PO4 and CuSO4 concentrations. The cultivation with a higher KH2PO4 concentration in a 400-mL bioreactor showed an increase in the maximum productivity of 1.88 g/L/h, which was an increase of 74 % in comparison to the reference. Neither the phosphate concentration nor the nitrogen sources were limited at the start of the product formation. This showed that a limitation of these substances is not necessary for the itaconic acid formation.


Aspergillus terreus Itaconic acid Cultivation Microtiter plate Optimization 



Many thanks for the financial support by the Fachagentur Nachwachsende Rohstoffe e.V. (grant no. 22020908).


  1. Calam CT, Oxford AE, Raistrick H (1939) Studies in the biochemistry of micro-organisms: itaconic acid, a metabolic product of a strain of Aspergillus terreus. Thom Biochem J 33(9):1488Google Scholar
  2. Futamura T, Okabe M, Tamura T, Toda K, Matsunobu T, Park YS (2001) Improvement of production of kojic acid by a mutant strain Aspergillus oryzae, MK107-39. J Biosci Bioeng 91(3):272–276PubMedGoogle Scholar
  3. Guevarra ED, Tabuchi T (1990) Accumulation of itaconic, 2-hydroxyparaconic, itatartaric, and malic acids by strains of the genus Ustilago (microbiology & fermentation industry). Agric Biol Chem 54(9):2353–2358CrossRefGoogle Scholar
  4. Gyamerah M (1995) Factors affecting the growth form of Aspergillus terreus NRRL 1960 in relation to itaconic acid fermentation. Appl Microbiol Biotechnol 44:356–361CrossRefGoogle Scholar
  5. Jahn B, Martin E, Stueben A, Bhakdi S (1995) Susceptibility testing of Candida albicans and Aspergillus species by a simple microtiter menadione-augmented 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay. J Clin Microbiol 33(3):661–667PubMedCentralPubMedGoogle Scholar
  6. Kuenz A, Gallenmüller Y, Willke T, Vorlop K-D (2012) Microbial production of itaconic acid: developing a stable platform for high product concentrations. Appl Microbiol Biotechnol 96(5):1209–1216PubMedCrossRefGoogle Scholar
  7. Larsen H, Eimhjellen K (1955) The mechanism of itaconic acid formation by Aspergillus terreus. 1. The effect of acidity. Biochem J 60(1):135PubMedCentralPubMedGoogle Scholar
  8. Levinson WE, Kurtzman CP, Kuo TM (2006) Production of itaconic acid by Pseudozyma antarctica NRRL Y-7808 under nitrogen-limited growth conditions. Enzym Microb Technol 39(4):824–827CrossRefGoogle Scholar
  9. Lockwood LB, Reeves MD (1945) Some factors affecting the production of itaconic acid by Aspergillus terreus. Arch Biochem 6:455–469Google Scholar
  10. Okabe M, Lies D, Kanamasa S, Park EY (2009) Biotechnological production of itaconic acid and its biosynthesis in Aspergillus terreus. Appl Microbiol Biotechnol 84(4):597–606PubMedCrossRefGoogle Scholar
  11. Papagianni M (2004) Fungal morphology and metabolite production insubmerged mycelial processes. Biotechnol Adv 22:189–259PubMedCrossRefGoogle Scholar
  12. Riscaldati E, Moresi M, Federici F, Petruccioli M (2000) Effect of pH and stirring rate on itaconate production by Aspergillus terreus. J Biotechnol 83:219–230PubMedCrossRefGoogle Scholar
  13. Rychtera M, Wase D (1981) The growth of Aspergillus terreus and the production of itaconic acid in batch and continuous cultures. The influence of pH. J Chem Technol Biotechnol 31(1):509–521CrossRefGoogle Scholar
  14. Tabuchi T, Sugisawa T, Ishidori T, Nakahara T, Sugiyama J (1981) Itaconic acid fermentation by a yeast belonging to the genus Candida. Agric Biol Chem 45(2):475–479CrossRefGoogle Scholar
  15. Tawara S, Ikeda F, Maki K, Morishita Y, Otomo K, Teratani N, Goto T, Tomishima M, Ohki H, Yamada A (2000) In vitro activities of a new lipopeptide antifungal agent, FK463, against a variety of clinically important fungi. Antimicrob Agents Chemother 44(1):57–62PubMedCentralPubMedCrossRefGoogle Scholar
  16. Willke T, Vorlop K-D (2001) Biotechnological production of itaconic acid. Appl Microbiol Biotechnol 56(3–4):289–295PubMedCrossRefGoogle Scholar
  17. Yahiro K, Takahama T, Jai SR, Park YS, Okabe M (1997) Comparison of air-lift and stirred tank reactors for itaconic acid production by Aspergillus terreus. Biotechnol Lett 19:619–621CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Antje Hevekerl
    • 1
  • Anja Kuenz
    • 1
  • Klaus-Dieter Vorlop
    • 1
  1. 1.Thünen-Institute of Agricultural TechnologyBraunschweigGermany

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