Rapid reagentless quantification of alginate biosynthesis in Pseudomonas fluorescens bacteria mutants using FT-IR spectroscopy coupled to multivariate partial least squares regression

Elon Correa, Havard Sletta, David I. Ellis, Sunniva Hoel, Helga Ertesvag, Trond E. Ellingsen, Svein Valla, Royston Goodacre

    Research output: Contribution to journalArticlepeer-review

    Abstract

    Alginate is an important medical and commercial product and currently is isolated from seaweeds. Certain microorganisms also produce alginate and these polymers have the potential to replace seaweed alginates in some applications, mainly because such production will allow much better and more reproducible control of critical qualitative polymer properties. The research conducted here presents the development of a new approach to this problem by analysing a transposon insertion mutant library constructed in an alginate-producing derivative of the Pseudomonas fluorescens strain SBW25. The procedure is based on the non-destructive and reagent-free method of Fourier transform infrared (FT-IR) spectroscopy which is used to generate a complex biochemical infrared fingerprint of the medium after bacterial growth. First, we investigate the potential differences caused by the growth media fructose and glycerol on the bacterial phenotype and alginate synthesis in 193 selected P. fluorescens mutants and show that clear phenotypic differences are observed in the infrared fingerprints. In order to quantify the level of the alginate we also report the construction and interpretation of multivariate partial least squares regression models which were able to quantify alginate levels successfully with typical normalized root-mean-square error in predictions of only approximately 14 %. We have demonstrated that this high-throughput approach can be implemented in alginate screens and we believe that this FT-IR spectroscopic methodology, when combined with the most appropriate chemometrics, could easily be modified for the quantification of other valuable microbial products and play a valuable screening role for synthetic biology.
    Original languageEnglish
    Pages (from-to)2591-2599
    Number of pages9
    JournalAnalytical and bioanalytical chemistry
    Volume403
    Issue number9
    DOIs
    Publication statusPublished - Jul 2012

    Keywords

    • Alginate
    • Alginate acetylation
    • FT-IR spectroscopy
    • Partial least squares regression
    • Pseudomonas fluorescens

    Research Beacons, Institutes and Platforms

    • Manchester Institute of Biotechnology

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