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Identification of lipophilic bioproduct portfolio from bioreactor samples of extreme halophilic archaea with HPLC-MS/MS

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Abstract

Extreme halophilic archaea are a yet unexploited source of natural carotenoids. At elevated salinities, however, material corrosivity issues occur and the performance of analytical methods is strongly affected. The goal of this study was to develop a method for identification and downstream processing of potentially valuable bioproducts produced by archaea. To circumvent extreme salinities during analysis, a direct sample preparation method was established to selectively extract both the polar and the nonpolar lipid contents of extreme halophiles with hexane, acetone and the mixture of MeOH/MTBE/water, respectively. Halogenated solvents, as used in conventional extraction methods, were omitted because of environmental considerations and potential process scale-up. The HPLC-MS/MS method using atmospheric pressure chemical ionization was developed and tuned with three commercially available C40 carotenoid standards, covering the wide polarity range of natural carotenoids, containing different number of OH-groups. The chromatographic separation was achieved on a C30 RP-HPLC column with a MeOH/MTBE/water gradient. Polar lipids, the geometric isomers of the C50 carotenoid bacterioruberin, and vitamin MK-8 were the most valuable products found in bioreactor samples. In contrast to literature on shake flask cultivations, no anhydrous analogues of bacterioruberin, as by-products of the carotenoid biosynthesis, were detected in bioreactor samples. This study demonstrates the importance of sample preparation and the applicability of HPLC-MS/MS methods on real samples from extreme halophilic strains. Furthermore, from a biotechnological point-of-view, this study would like to reveal the relevance of using controlled and defined bioreactor cultivations instead of shake flask cultures in the early stage of potential bioproduct profiling.

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Acknowledgments

C. Koch and B. Lendl gratefully acknowledge partial financial support by the Austrian research funding association (FFG) under the scope of the COMET program within the research network Process Analytical Chemistry (PAC) (contract no. 825340). This program is promoted by BMVIT (Federal Ministry of Transport and Innovation), BMWFJ (Federal Ministry of Economy, Family, and Youth), and the federal state of upper Austria. Thanks are expressed to Bernhard Seyer for the biomass samples. The authors thank the Referees for their very helpful comments.

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Authors and Affiliations

  1. Research Division Biochemical Engineering, Institute of Chemical Engineering, Vienna University of Technology, Gumpendorferstrasse 1A 166/4, 1060, Vienna, Austria

    Bettina Lorantfy & Christoph Herwig

  2. Research Center Pharmaceutical Engineering, Inffeldgasse 21a, 8010, Graz, Austria

    Bettina Lorantfy

  3. Department of Inorganic and Analytical Chemistry, Budapest University of Technology and Economics, Szent Gellért tér 4, 1111, Budapest, Hungary

    Tibor Renkecz & George Horvai

  4. MTA-BME Research Group of Technical Analytical Chemistry, Szent Gellért tér 4, 1111, Budapest, Hungary

    George Horvai

  5. Vienna University of Technology, Institute of Chemical Technologies and Analytics, Getreidemarkt 9 164-UPA, 1060, Vienna, Austria

    Cosima Koch & Bernhard Lendl

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  1. Bettina Lorantfy
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  2. Tibor Renkecz
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Correspondence to Christoph Herwig.

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Lorantfy, B., Renkecz, T., Koch, C. et al. Identification of lipophilic bioproduct portfolio from bioreactor samples of extreme halophilic archaea with HPLC-MS/MS. Anal Bioanal Chem 406, 2421–2432 (2014). https://doi.org/10.1007/s00216-014-7626-x

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