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. (Kuenz, 2014)

In this study, a systematic process optimization was performed with an own isolated strain of Aspergillus
terreus and transferred from a 250-mL to a 15-L scale. An IA concentration of 86.2 g/L was achieved
within 7 days with an overall productivity of 0.51 g/(L h), a maximum productivity of 1.2 g/(L h), and a
yield of 86 mol%. (Kuenz, 2012)

The LSF in optimized culture medium confirmed the ability of A. terreus strains to produce IA. A. terreus
826 produced 90 g/l of IA in a totally controlled bioreactor with 180 g/l of glucose, resulting in a yield of
0.5 g IA/g total glucose. (Kuenz, 2012)

Steam explosion is a successful pretreatment option that involves heating lignocellulose with
superheated steam followed by a sudden decompression. The high-pressure steam modifies the cell wall
structure, yielding a slurry, which upon filtration renders a filtrate with hemicellulosic sugars and a
cellulose-rich filter cake containing also lignin and residual hemicellulose. Steam explosion can be
assisted by impregnation with an acid catalyst, for instance sulfuric acid or sulfur dioxide. If no
impregnating agent is used, the process is catalyzed through autohydrolysis. Acetic acid and uronic acids
released from hemicellulose, and formic and levulinic acids resulting from sugar degradation (Fig. 1)
contribute to acidification, and can inhibit downstream biochemical processes

A study conducted by Yang et al. (2011) obtained a similar cellulose conversion (70.6%), with a higher
solids loading (30%), an enzyme loading almost twice (20 FPU/g cellulose) that used in the former study
and with a much shorter reaction time (30 hrs). Both studies attribute the high conversion rate, at least
in part, to the fact that the substrates were washed prior to hydrolysis, possibly eliminating any
potential inhibitory products that resulted from the pretreatments.

Aytac, K., B.O. Zumrut and B. Ufuk, 2014. Xylanase and itaconic acid production by Aspergillus terreus
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Bentley, R. and C.P. Thiessen, 1957. Biosynthesis of itaconic acid in Aspergillusterreus. I. Tracer studies
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Jarry, A. and Y. Seraudie, 1995. Production of Itaconic Acid by Fermentation. US Patent No. 5.457.040.

Kautola, H., M. Vahvaselka, Y.Y. Linko and P. Linko, 1985. Itaconic acid production by immobilized
Aspergillus terreus from xylose and glucose. Biotechnol. Lett., 7: 167-172.

Mitsuyasu, O., L. Dwiarti, K. Shin and P.Y. Enoch, 2009. Biotechnological production of itaconic acid and
its biosynthesis in Aspergillus terreus. Appl. Microbiol. Biotechnol., 84: 597-606.
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biosynthesis in Aspergillus terreus. Appl. Microbiol. Biotechnol., 84: 597-606.

Park, Y., M. Itida, N. Ohta and M. Okabe, 1994. Itaconic acid production using an air-lift bioreactor in
repeated batch culture of Aspergillus terreus. J. Ferment. Bioeng., 77: 329-331.

Tsai, Y., M. Huang, S. Lin and Y. Su, 2001. Method for the Production of Itaconic Acid Using Aspergillus
terreus Solid State Fermentation. US Patent No. 6,171,831.

Van der Werf, M., M. Caspers and M. Petrus, 2009. Production of itaconic acid. EP Patent No. 017344
A1.

Yahiro, K., T. Takahama, Y. Park and M. Okabe, 1995. Breeding of Aspergillus terreus Mutant TN-484 for
an itaconic acid production with high yield. J. Ferm. Bioeng., 79: 506-508.

Production culture containing glucose as sole carbon source was used as control for IA production (Fig.
1a) and medium including 1 % (w/v) birchwood xylan instead of glucose was used for xylanase
production (Fig. 1b)

Willke and Vorlop (2001) claimed that 100–150 g/L glucose was required for a satisfactory production
rate and amount of production of IA from A. terreus. Hence, to prevent consumption of IA by the
organism, a high concentration of simple sugar should be supplied throughout the whole fermentation
period