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

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

doi:10.1186/s12934-017-0716-7

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

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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 (RPD_CV = 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 language	English
Article number	101
Journal	Microbial Cell Factories
Volume	16
Issue number	1
DOIs	https://doi.org/10.1186/s12934-017-0716-7
Publication status	Published - 9 Jun 2017
MoE publication type	A1 Journal article-refereed

Keywords

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

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title = "Microtiter plate cultivation of oleaginous fungi and monitoring of lipogenesis by high-throughput FTIR spectroscopy",

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.",

keywords = "Fatty acid analysis, GC-FID, High-throughput FTIR spectroscopy, Microcultivation, Oleaginous fungi, PLS regression",

author = "Gergely Kosa and Achim Kohler and Valeria Tafintseva and Boris Zimmermann and Kristin Forfang and Afseth, {Nils Kristian} and Dimitrios Tzimorotas and Vuoristo, {Kiira S.} and Horn, {Svein Jarle} and Jerome Mounier and Volha Shapaval",

note = "Funding Information: This work was supported by the Norwegian Research Council-BION{\AE}R Grant, project number: 234258/E50 and the Norwegian Research Council “Interest” Grant, project number 227356. Publisher Copyright: {\textcopyright} 2017 The Author(s).",

year = "2017",

month = jun,

day = "9",

doi = "10.1186/s12934-017-0716-7",

language = "English",

volume = "16",

journal = "Microbial Cell Factories",

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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

N1 - Funding Information: This work was supported by the Norwegian Research Council-BIONÆR Grant, project number: 234258/E50 and the Norwegian Research Council “Interest” Grant, project number 227356. Publisher Copyright: © 2017 The Author(s).

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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U2 - 10.1186/s12934-017-0716-7

DO - 10.1186/s12934-017-0716-7

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SN - 1475-2859

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ER -

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

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Keywords

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