In situ detection of aromatic compounds with biosensor Pseudomonas putida cells preserved and delivered to soil in water-soluble gelatin capsules

Abstract

While many types of bacteria have been engineered to produce an optical output in response to given analytes in a culture, their use for extensive, in situ monitoring of distinct chemical species in soil is hampered by a dearth of practicable spreading schemes. In this work, we report and validate a comprehensive system for the long-term preservation of Pseudomonas putida cells genetically designed for biosensing benzene, toluene, ethylbenzene, and xylenes (BTEX) in soil, along with a procedure to formulate, spread, and vigorously activate such bacteria at the desired site and occasion. To this end, various known lyoprotectants were tested for promoting the long-term maintenance of biosensor cells with quite variable outcomes. While a formulation of inositol and maltodextrines was optimal for preservation of freeze-dried BTEX-sensing bacteria, adsorption of P. putida cells to corncob powder (an abundant residue of the corn industry) endowed the resulting material with a lasting viability at ambient conditions. In any case, the thereby preserved bacterial biomass acquired physical and mechanical properties adequate for formulating the biosensor agent in water-soluble but otherwise hard dry gelatine capsules with a long shelf life. When such capsules were spread in a soil microcosm and subsequently liquefied with water or high humidity, the released microorganisms formed spots that gave an intense luminiscent signal upon exposure to effectors of the sensor circuit implanted in the chromosome of the P. putida strain. We argue that the procedures described here can facilitate implementation of wide-area biological detection strategies for revealing the location of toxic or perilous chemicals.

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Notes

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    This strain was certified as early as in 1981 by the Recombinant DNA Advisory Committee of the NIH as the first host-vector biosafety system for gene cloning in Gram-negative soil bacteria (Federal Register of 1982, Appendix E, Certified Host-Vector Systems, vol. 47, 17197).

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Acknowledgment

We thank JR van der Meer (University of Lausanne, Switzerland) for his gift of corncob and J. Pliego (OMFE) for providing hard gelatin capsules. We also thank Rob Cain for his comments and Esther Fernández for technical help. This work was defrayed by generous grants of the CONSOLIDER program of the Spanish Ministry of Science and Innovation, by the TARPOL, BACSINE, and MICROME Contracts of the EU and by funds of the Autonomous Community of Madrid.

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Correspondence to Víctor de Lorenzo.

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Published in the special issue Microorganisms for Analysis with Guest Editor Gérald Thouand.

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de las Heras, A., de Lorenzo, V. In situ detection of aromatic compounds with biosensor Pseudomonas putida cells preserved and delivered to soil in water-soluble gelatin capsules. Anal Bioanal Chem 400, 1093–1104 (2011). https://doi.org/10.1007/s00216-010-4558-y

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Keywords

  • Pseudomonas putida
  • XylR
  • Biosensors
  • Desiccation
  • Encapsulation