Abstract
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.
Similar content being viewed by others
References
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):1488
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):272–276
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):2353–2358
Gyamerah M (1995) Factors affecting the growth form of Aspergillus terreus NRRL 1960 in relation to itaconic acid fermentation. Appl Microbiol Biotechnol 44:356–361
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):661–667
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):1209–1216
Larsen H, Eimhjellen K (1955) The mechanism of itaconic acid formation by Aspergillus terreus. 1. The effect of acidity. Biochem J 60(1):135
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):824–827
Lockwood LB, Reeves MD (1945) Some factors affecting the production of itaconic acid by Aspergillus terreus. Arch Biochem 6:455–469
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):597–606
Papagianni M (2004) Fungal morphology and metabolite production insubmerged mycelial processes. Biotechnol Adv 22:189–259
Riscaldati E, Moresi M, Federici F, Petruccioli M (2000) Effect of pH and stirring rate on itaconate production by Aspergillus terreus. J Biotechnol 83:219–230
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):509–521
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):475–479
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):57–62
Willke T, Vorlop K-D (2001) Biotechnological production of itaconic acid. Appl Microbiol Biotechnol 56(3–4):289–295
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 19:619–621
Acknowledgments
Many thanks for the financial support by the Fachagentur Nachwachsende Rohstoffe e.V. (grant no. 22020908).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Hevekerl, A., Kuenz, A. & Vorlop, KD. Filamentous fungi in microtiter plates—an easy way to optimize itaconic acid production with Aspergillus terreus . Appl Microbiol Biotechnol 98, 6983–6989 (2014). https://doi.org/10.1007/s00253-014-5743-2
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-014-5743-2