Bioreactor as the root cause of the "manganese effect" during Aspergillus niger citric acid fermentations

Erzsébet Fekete; Vivien Bíró; Alexandra Márton; István Bakondi-Kovács; Zoltán Németh; Erzsébet Sándor; Béla Kovács; István Fábián; Christian P Kubicek; Adrian Tsang; Levente Karaffa

doi:10.3389/fbioe.2022.935902

Bioreactor as the root cause of the "manganese effect" during Aspergillus niger citric acid fermentations

Front Bioeng Biotechnol. 2022 Aug 4:10:935902. doi: 10.3389/fbioe.2022.935902. eCollection 2022.

Authors

Erzsébet Fekete¹, Vivien Bíró^{1

2}, Alexandra Márton^{1

2}, István Bakondi-Kovács^{1

2}, Zoltán Németh¹, Erzsébet Sándor³, Béla Kovács³, István Fábián^{4

5}, Christian P Kubicek⁶, Adrian Tsang⁷, Levente Karaffa^{1

8}

Affiliations

¹ Department of Biochemical Engineering, Faculty of Science and Technology, University of Debrecen, Debrecen, Hungary.
² Juhász-Nagy Pál Doctoral School of Biology and Environmental Sciences, University of Debrecen, Debrecen, Hungary.
³ Institute of Food Science, Faculty of Agricultural and Food Science and Environmental Management, University of Debrecen, Debrecen, Hungary.
⁴ Department of Inorganic and Analytical Chemistry, Faculty of Science and Technology, University of Debrecen, Debrecen, Hungary.
⁵ MTA-DE Redox and Homogeneous Catalytic Reaction Mechanism Research Group, Debrecen, Hungary.
⁶ Institute of Chemical, Environmental and Bioscience Engineering, TU Wien, Vienna, Austria.
⁷ Centre for Structural and Functional Genomics, Concordia University, Montreal, QC, Canada.
⁸ Institute of Metagenomics, University of Debrecen, Debrecen, Hungary.

Abstract

High-yield citric acid production by the filamentous Ascomycete fungus Aspergillus niger requires a combination of extreme nutritional conditions, of which maintaining a low manganese (II) ion concentration (<5 μg L^-1) is a key feature. Technical-scale production of citric acid predominantly uses stainless-steel tank fermenters, but glass bioreactors used for strain improvement and manufacturing process development also contain stainless steel components, in which manganese is an essential alloying element. We show here that during citric acid fermentations manganese (II) ions were leaching from the bioreactor into the growth media, resulting in altered fungal physiology and morphology, and significant reduction of citric acid yields. The leaching of manganese (II) ions was dependent on the fermentation time, the acidity of the culture broth and the sterilization protocol applied. Manganese (II) ion leaching was partially mitigated by electrochemical polishing of stainless steel components of the bioreactor. High concentrations of manganese (II) ions during early cultivation led to a reduction in citric acid yield. However, the effect of manganese (II) ions on the reduction of citric acid yield diminished towards the second half of the fermentation. Since maintaining low concentrations of manganese (II) ions is costly, the results of this study can potentially be used to modify protocols to reduce the cost of citric acid production.

Keywords: Aspergillus niger; citric acid; fungal morphology; manganese ions; metal ions leaching; stainless steel.