Dataset Information

Metabolite Quantification by Fourier Transform Infrared Spectroscopy in Diatoms: Proof of Concept on Phaeodactylum tricornutum.

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.

SUBMITTER: Scarsini M

PROVIDER: S-EPMC8631545 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature

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Publications

Metabolite Quantification by Fourier Transform Infrared Spectroscopy in Diatoms: Proof of Concept on <i>Phaeodactylum tricornutum</i>.

Scarsini Matteo M Thurotte Adrien A Veidl Brigitte B Amiard Frederic F Niepceron Frederick F Badawi Myriam M Lagarde Fabienne F Schoefs Benoît B Marchand Justine J

Frontiers in plant science 20211110

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 ...[more]

PMID: 34858459

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Project description:Fourier transform infrared (FTIR) spectroscopy, a technology traditionally used in chemistry to determine the molecular composition of a wide range of sample types, has gained growing interest in microbial typing. It is based on the different vibrational modes of the covalent bonds between atoms of a given sample, as bacterial cells, induced by the absorption of infrared radiation. This technique has been largely used for the study of pathogenic species, especially in the clinical field, and has been proposed also for the typing at different subspecies levels. The high throughput, speed, low cost, and simplicity make FTIR spectroscopy an attractive technique also for industrial applications, in particular, for probiotics. The aim of this study was to compare FTIR spectroscopy with established genotyping methods, pulsed-field gel electrophoresis (PFGE), whole-genome sequencing (WGS), and multilocus sequence typing (MLST), in order to highlight the FTIR spectroscopy potential discriminatory power at strain level. Our study focused on bifidobacteria, an important group of intestinal commensals generally recognized as probiotics. For their properties in promoting and maintaining health, bifidobacteria are largely marketed by the pharmaceutical, food, and dairy industries. Strains belonging to Bifidobacterium longum subsp. longum and Bifidobacterium animalis subsp. lactis were taken into consideration together with some additional type strains. For B. longum subsp. longum, it was possible to discriminate the strains with all the methods used. Although two isolates were shown to be strictly phylogenetically related, constituting a unique cluster, based on PFGE, WGS, and MLST, no clustering was observed with FTIR. For B. animalis subsp. lactis group, PFGE, WGS, and MLST were non-discriminatory, and only one strain was easily distinguished. On the other hand, FTIR discriminated all the isolates one by one, and no clustering was observed. According to these results, FTIR analysis is not only equivalent to PFGE, WGS, and MLST, but also for some strains, in particular, for B. animalis subsp. lactis group, more informative, being able to differentiate strains not discernible with the other two methods based on phenotypic variations likely deriving from certain genetic changes. Fourier transform infrared spectroscopy has highlighted the possibility of using the cell surface as a kind of barcode making tracing strains possible, representing an important aspect in probiotic applications. Furthermore, this work constitutes the first investigation on bifidobacterial strain typing using FTIR spectroscopy.

Dataset Information

Metabolite Quantification by Fourier Transform Infrared Spectroscopy in Diatoms: Proof of Concept on Phaeodactylum tricornutum.

Publications

Metabolite Quantification by Fourier Transform Infrared Spectroscopy in Diatoms: Proof of Concept on <i>Phaeodactylum tricornutum</i>.

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