Microtiter plate cultivation of oleaginous fungi and monitoring of lipogenesis by high-throughput FTIR spectroscopy

Gergely Kosa (Corresponding Author), Achim Kohler, Valeria Tafintseva, Boris Zimmermann, Kristin Forfang, Nils Kristian Afseth, Dimitrios Tzimorotas, Kiira S. Vuoristo, Svein Jarle Horn, Jerome Mounier, Volha Shapaval

Research output: Contribution to journalArticleScientificpeer-review

15 Citations (Scopus)

Abstract

Background: Oleaginous fungi can accumulate lipids by utilizing a wide range of waste substrates. They are an important source for the industrial production of omega-6 polyunsaturated fatty acids (gamma-linolenic and arachidonic acid) and have been suggested as an alternative route for biodiesel production. Initial research steps for various applications include the screening of fungi in order to find efficient fungal producers with desired fatty acid composition. Traditional cultivation methods (shake flask) and lipid analysis (extraction-gas chromatography) are not applicable for large-scale screening due to their low throughput and time-consuming analysis. Here we present a microcultivation system combined with high-throughput Fourier transform infrared (FTIR) spectroscopy for efficient screening of oleaginous fungi. Results: The microcultivation system enables highly reproducible fungal fermentations throughout 12 days of cultivation. Reproducibility was validated by FTIR and HPLC data. Analysis of FTIR spectral ester carbonyl peaks of fungal biomass offered a reliable high-throughput at-line method to monitor lipid accumulation. Partial least square regression between gas chromatography fatty acid data and corresponding FTIR spectral data was used to set up calibration models for the prediction of saturated fatty acids, monounsaturated fatty acids, polyunsaturated fatty acids, unsaturation index, total lipid content and main individual fatty acids. High coefficients of determination (R2 = 0.86-0.96) and satisfactory residual predictive deviation of cross-validation (RPDCV = 2.6-5.1) values demonstrated the goodness of these models. Conclusions: We have demonstrated in this study, that the presented microcultivation system combined with rapid, high-throughput FTIR spectroscopy is a suitable screening platform for oleaginous fungi. Sample preparation for FTIR measurements can be automated to further increase throughput of the system.

Original languageEnglish
Article number101
JournalMicrobial Cell Factories
Volume16
Issue number1
DOIs
Publication statusPublished - 9 Jun 2017
MoE publication typeA1 Journal article-refereed

Fingerprint

Lipogenesis
Fourier Analysis
Fourier Transform Infrared Spectroscopy
Fungi
Fourier transform infrared spectroscopy
Fatty Acids
Lipids
Throughput
Fourier transforms
Screening
Fatty acids
Monitoring
Polyunsaturated fatty acids
Infrared radiation
Unsaturated Fatty Acids
Gas Chromatography
Gas chromatography
Omega-6 Fatty Acids
Monounsaturated fatty acids
Monounsaturated Fatty Acids

Keywords

  • Fatty acid analysis
  • GC-FID
  • High-throughput FTIR spectroscopy
  • Microcultivation
  • Oleaginous fungi
  • PLS regression

Cite this

Kosa, G., Kohler, A., Tafintseva, V., Zimmermann, B., Forfang, K., Afseth, N. K., ... Shapaval, V. (2017). Microtiter plate cultivation of oleaginous fungi and monitoring of lipogenesis by high-throughput FTIR spectroscopy. Microbial Cell Factories, 16(1), [101]. https://doi.org/10.1186/s12934-017-0716-7
Kosa, Gergely ; Kohler, Achim ; Tafintseva, Valeria ; Zimmermann, Boris ; Forfang, Kristin ; Afseth, Nils Kristian ; Tzimorotas, Dimitrios ; Vuoristo, Kiira S. ; Horn, Svein Jarle ; Mounier, Jerome ; Shapaval, Volha. / Microtiter plate cultivation of oleaginous fungi and monitoring of lipogenesis by high-throughput FTIR spectroscopy. In: Microbial Cell Factories. 2017 ; Vol. 16, No. 1.
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Kosa, G, Kohler, A, Tafintseva, V, Zimmermann, B, Forfang, K, Afseth, NK, Tzimorotas, D, Vuoristo, KS, Horn, SJ, Mounier, J & Shapaval, V 2017, 'Microtiter plate cultivation of oleaginous fungi and monitoring of lipogenesis by high-throughput FTIR spectroscopy', Microbial Cell Factories, vol. 16, no. 1, 101. https://doi.org/10.1186/s12934-017-0716-7

Microtiter plate cultivation of oleaginous fungi and monitoring of lipogenesis by high-throughput FTIR spectroscopy. / Kosa, Gergely (Corresponding Author); Kohler, Achim; Tafintseva, Valeria; Zimmermann, Boris; Forfang, Kristin; Afseth, Nils Kristian; Tzimorotas, Dimitrios; Vuoristo, Kiira S.; Horn, Svein Jarle; Mounier, Jerome; Shapaval, Volha.

In: Microbial Cell Factories, Vol. 16, No. 1, 101, 09.06.2017.

Research output: Contribution to journalArticleScientificpeer-review

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T1 - Microtiter plate cultivation of oleaginous fungi and monitoring of lipogenesis by high-throughput FTIR spectroscopy

AU - Kosa, Gergely

AU - Kohler, Achim

AU - Tafintseva, Valeria

AU - Zimmermann, Boris

AU - Forfang, Kristin

AU - Afseth, Nils Kristian

AU - Tzimorotas, Dimitrios

AU - Vuoristo, Kiira S.

AU - Horn, Svein Jarle

AU - Mounier, Jerome

AU - Shapaval, Volha

PY - 2017/6/9

Y1 - 2017/6/9

N2 - Background: Oleaginous fungi can accumulate lipids by utilizing a wide range of waste substrates. They are an important source for the industrial production of omega-6 polyunsaturated fatty acids (gamma-linolenic and arachidonic acid) and have been suggested as an alternative route for biodiesel production. Initial research steps for various applications include the screening of fungi in order to find efficient fungal producers with desired fatty acid composition. Traditional cultivation methods (shake flask) and lipid analysis (extraction-gas chromatography) are not applicable for large-scale screening due to their low throughput and time-consuming analysis. Here we present a microcultivation system combined with high-throughput Fourier transform infrared (FTIR) spectroscopy for efficient screening of oleaginous fungi. Results: The microcultivation system enables highly reproducible fungal fermentations throughout 12 days of cultivation. Reproducibility was validated by FTIR and HPLC data. Analysis of FTIR spectral ester carbonyl peaks of fungal biomass offered a reliable high-throughput at-line method to monitor lipid accumulation. Partial least square regression between gas chromatography fatty acid data and corresponding FTIR spectral data was used to set up calibration models for the prediction of saturated fatty acids, monounsaturated fatty acids, polyunsaturated fatty acids, unsaturation index, total lipid content and main individual fatty acids. High coefficients of determination (R2 = 0.86-0.96) and satisfactory residual predictive deviation of cross-validation (RPDCV = 2.6-5.1) values demonstrated the goodness of these models. Conclusions: We have demonstrated in this study, that the presented microcultivation system combined with rapid, high-throughput FTIR spectroscopy is a suitable screening platform for oleaginous fungi. Sample preparation for FTIR measurements can be automated to further increase throughput of the system.

AB - Background: Oleaginous fungi can accumulate lipids by utilizing a wide range of waste substrates. They are an important source for the industrial production of omega-6 polyunsaturated fatty acids (gamma-linolenic and arachidonic acid) and have been suggested as an alternative route for biodiesel production. Initial research steps for various applications include the screening of fungi in order to find efficient fungal producers with desired fatty acid composition. Traditional cultivation methods (shake flask) and lipid analysis (extraction-gas chromatography) are not applicable for large-scale screening due to their low throughput and time-consuming analysis. Here we present a microcultivation system combined with high-throughput Fourier transform infrared (FTIR) spectroscopy for efficient screening of oleaginous fungi. Results: The microcultivation system enables highly reproducible fungal fermentations throughout 12 days of cultivation. Reproducibility was validated by FTIR and HPLC data. Analysis of FTIR spectral ester carbonyl peaks of fungal biomass offered a reliable high-throughput at-line method to monitor lipid accumulation. Partial least square regression between gas chromatography fatty acid data and corresponding FTIR spectral data was used to set up calibration models for the prediction of saturated fatty acids, monounsaturated fatty acids, polyunsaturated fatty acids, unsaturation index, total lipid content and main individual fatty acids. High coefficients of determination (R2 = 0.86-0.96) and satisfactory residual predictive deviation of cross-validation (RPDCV = 2.6-5.1) values demonstrated the goodness of these models. Conclusions: We have demonstrated in this study, that the presented microcultivation system combined with rapid, high-throughput FTIR spectroscopy is a suitable screening platform for oleaginous fungi. Sample preparation for FTIR measurements can be automated to further increase throughput of the system.

KW - Fatty acid analysis

KW - GC-FID

KW - High-throughput FTIR spectroscopy

KW - Microcultivation

KW - Oleaginous fungi

KW - PLS regression

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