Fourier transform infrared spectroscopy as a method to study lipid accumulation in oleaginous yeasts
Open accessAbstract
BACKGROUND: Oleaginous microorganisms, such as different yeast and algal species, can represent a sustainable alternative to plant oil for the production of biodiesel. They can accumulate fatty acids (FA) up to 70% of their dry weight with a predominance of (mono)unsaturated species, similarly to what plants do, but differently from animals. In addition, their growth is not in competition either with food, feed crops, or with agricultural land.Despite these advantages, the exploitation of the single cell oil system is still at an early developmental stage. Cultivation mode and conditions, as well as lipid extraction technologies, represent the main limitations. The monitoring of lipid accumulation in oleaginous microorganisms is consequently crucial to develop and validate new approaches, but at present the majority of the available techniques is time consuming, invasive and, when relying on lipid extraction, can be affected by FA degradation. RESULTS: In this work the fatty acid accumulation of the oleaginous yeasts Cryptococcus curvatus and Rhodosporidium toruloides and of the non-oleaginous yeast Saccharomyces cerevisiae (as a negative control) was monitored in situ by Fourier Transform Infrared Spectroscopy (FTIR). Indeed, this spectroscopic tool can provide complementary information to those obtained by classical techniques, such as microscopy, flow cytometry and gas chromatography. As shown in this work, through the analysis of the absorption spectra of intact oleaginous microorganisms it is possible not only to monitor the progression of FA accumulation but also to identify the most represented classes of the produced lipids. CONCLUSIONS: Here we propose FTIR microspectroscopy - supported by multivariate analysis - as a fast, reliable and non invasive method to monitor and analyze FA accumulation in intact oleaginous yeasts. The results obtained by the FTIR approach were in agreement with those obtained by the other classical methods like flow cytometry and gas chromatography. Moreover, the possibility to track lipid production in real time is highly desirable to support the initial screening of strains and media as well as to optimize the scaling up experiments, which are essential for a viable and successful development of an industrial production process.
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Second derivative spectroscopy enables accurate measurements of wavelength position by identifying small and overlapping peaks that are not resolved in the original spectra.
“SD spectroscopy enables accurate measurements of wavelength position by identifying small and overlapping peaks that are not resolved in the original spectra. [23,46,47].”
From Response to nutrient variation on lipid productivity in green microalgae captured using second derivative FTIR and Raman spectroscopy (2022) · cites this paper as [47]
Spectral intensity variation due to biomolecular transitions can be attributed to cell metabolism during growth phases.
“Thus the intensity variation observed in the spectra due to biomolecular transitions can be attributed to the metabolism of cells during their various phases of growth [26,28].”
From Biomolecular transitions and lipid accumulation in green microalgae monitored by FTIR and Raman analysis (2020) · cites this paper as [26]
The four characteristic hydrocarbon bands are at ~2960 (as CH3), ~2920 (as CH2), ~2870 (s CH3), and ~2850 cm−1 (s CH2).
“The characteristic bands of hydrocarbons are the four bands at ~ 2960cm -1 ( as CH 3 ), 2920cm -1 ( as CH 2 ), 2870 cm -1 ( s CH 3 ) and 2850 cm -1 ( s CH 2 ) [22,26].”
From Biomolecular transitions and lipid accumulation in green microalgae monitored by FTIR and Raman analysis (2020) · cites this paper as [26]
Bands at ~3008 cm−1 ν(=C-H) and ~1270 cm−1 δ(=C-H) are due to stretching and bending vibrations of unsaturated fatty acids in algal lipids.
“The smaller bands at ~3008 cm -1 ν (= C-H) and ~ 1270 cm -1 (= C-H) were due to the stretching and bending vibrations of unsaturated fatty acids in the algal lipids.[21,26,50].”
From Biomolecular transitions and lipid accumulation in green microalgae monitored by FTIR and Raman analysis (2020) · cites this paper as [26]
Second derivative analysis of FTIR absorption spectra resolved overlapping bands, removed baseline errors, identified small adjacent bands, and enabled quantitative analysis.
“Second derivatives resolved the overlapping bands in the raw spectra, removed the baseline errors and lead to the identification of small and adjacent absorption bands and enabled quantitative analysis [26,49].”
From Biomolecular transitions and lipid accumulation in green microalgae monitored by FTIR and Raman analysis (2020) · cites this paper as [26]
The ~3008 cm−1 ν(=C-H) band from unsaturated fatty acids was clearly visible in M. contortum.
“The band at ~ 3008 cm -1 corresponded to the stretching vibrations ν (= C-H) from the unsaturated fatty acids was clearly visible in M.contortum.(Fig. 4b to Fig. 4d) [21,26].”
From Biomolecular transitions and lipid accumulation in green microalgae monitored by FTIR and Raman analysis (2020) · cites this paper as [26]
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References 49 · Cited by 97
- Authors
- Ami, D., Posteri, R., Mereghetti, P., Porro, D., Doglia, S. M., & Branduardi, P.
- Affiliations
- National Interuniversity Consortium for the Physical Sciences of Matter (IT)
- University of Milano-Bicocca (IT)
- Italian Institute of Technology (IT)
- Corresponding author
- Doglia, Silvia Maria
- Venue
- Biotechnology for Biofuels
- Published
- 2014-01-23
- Volume
- 7
- Issue
- 1
- Pages
- 12-12
- ISSN
- 1754-6834 (ISSN-L)
- PMC
- PMC3923900
- OpenAlex
- W2107531885
- Cited by
- 97
- References
- 49
- Citations by year
- 2026: 6 · 2025: 2 · 2024: 6 · 2023: 8 · 2022: 9 · 2021: 15 · 2020: 8 · 2019: 14 · 2018: 8 · 2017: 6 · 2016: 8 · 2015: 5 · 2014: 2
- FWCI
- 2.31
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by
- Locations
- Biotechnology for Biofuels (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- BOA (University of Milano-Bicocca) (submittedVersion, cc-by)
- Funders
- Università degli Studi di Milano-Bicocca
- Regione Lombardia (15083/RCC)
- Topics
- Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Algal biology and biofuel production
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Spectroscopy Techniques in Biomedical and Chemical Research
- Physical Sciences › Chemistry › Analytical Chemistry › Spectroscopy and Chemometric Analyses
- Keywords
- oleaginous yeast, lipid accumulation, fatty acid accumulation, in situ monitoring, FTIR microspectroscopy, Cryptococcus curvatus, Rhodosporidium toruloides, Saccharomyces cerevisiae, biodiesel feedstock, single cell oil, non-invasive monitoring, multivariate analysis
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