Abstract
Diatoms are feedstock for the production of sustainable biocommodities, including biofuel. The biochemical characterization of newly isolated or genetically modified strains is seminal to identify the strains that display interesting features for both research and industrial applications. Biochemical quantification of organic macromolecules cellular quotas are time-consuming methodologies which often require large amount of biological sample. Vibrational spectroscopy is an essential tool applied in several fields of research. A Fourier transform infrared (FTIR) microscopy-based imaging protocol was developed for the simultaneous cellular quota quantification of lipids, carbohydrates, and proteins of the diatom Phaeodactylum tricornutum. The low amount of sample required for the quantification allows the high throughput quantification on small volume cultures. A proof of concept was performed (1) on nitrogen-starved experimental cultures and (2) on three different P. tricornutum wild-type strains. The results are supported by the observation in situ of lipid droplets by confocal and brightfield microscopy. The results show that major differences exist in the regulation of lipid metabolism between ecotypes of P. tricornutum.
Highlights
The increase of human populations, the reduction of farmland and the expansion of cities contribute to an accelerated depletion of natural resources to a point that finding alternative sources of natural resources, including biomolecules, is becoming crucial
A high-throughput sensitive methodology based on Fourier transform infrared (FTIR) was designed and evaluated on the marine diatom P. tricornutum
The results show that the organic macromolecules classes can be quantified exploiting the IR absorbance at their specific wavenumber after the generation of proper calibration curves
Summary
The increase of human populations, the reduction of farmland and the expansion of cities contribute to an accelerated depletion of natural resources to a point that finding alternative sources of natural resources, including biomolecules, is becoming crucial. Microalgae are a promising feedstock for the production of biomolecules belonging to the major organic macromolecules classes (Mimouni et al, 2012) while utilizing sun energy through the photosynthetic process (Scarsini et al, 2019). The current knowledge on microalgae does not allow the development of biotechnological sustainable processes from the ecological and financial point of views. Obtaining appropriate strains dedicated to a specific production is one of the major challenges. To reach this goal, two strategies may be applied: (1) the exploration of microalgal biodiversity to find appropriate strains with high industrial application potential and (2) the genetic modifications of wild-type microalgal strains to improve their productivity. The identification of cell lines, which possess the phenotype of interest among a mutagenized population, can be a tedious process for which it may be necessary to analyze hundreds or even thousands of transformants
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