2.50
Hdl Handle:
http://hdl.handle.net/10033/12985
Title:
Analysing traces of autoinducer-2 requires standardization of the Vibrio harveyi bioassay.
Authors:
Vilchez, Ramiro; Lemme, André; Thiel, Verena; Schulz, Stefan; Sztajer, Helena; Wagner-Döbler, Irene
Abstract:
Autoinducer-2 (furanosyl borate diester) is a biologically active compound whose role as a universal bacterial signalling molecule is currently under intense investigation. Because of its instability and the low concentrations of it found in biological samples, its detection relies at present on a bioassay that measures the difference in the timing of the luminescence of the Vibrio harveyi BB170 sensor strain with and without externally added AI-2. Here we systematically investigated which parameters affected the fold induction values of luminescence obtained in the bioassay and developed a modified protocol. Our experiments showed that growth and luminescence of V. harveyi BB170 are strongly influenced by trace elements. In particular, addition of Fe(3+) within a certain concentration range to the growth medium of the preinoculum culture improved the reproducibility and reduced the variance of the bioassay. In contrast, trace elements and vitamins introduced directly into the bioassay caused inhibitory effects. The initial density and luminescence of the sensor strain are very important and the values required for these parameters were defined. Borate interferes with the detection of AI-2 by giving false positive results. The response of V. harveyi BB170 to chemically synthesized AI-2 in the bioassay is nonlinear except over a very small concentration range; it is maximum over three orders of magnitude and shows inhibition above 35 microM. Based on the modified protocol, we were able to detect AI-2 in the absence of inhibitors with maximum fold induction values for the positive control (chemically synthesized AI-2) of >120 with a standard deviation of approximately 30% in a reliable and reproducible way.
Citation:
Anal Bioanal Chem 2007, 387(2):489-96
Issue Date:
1-Jan-2007
URI:
http://hdl.handle.net/10033/12985
DOI:
10.1007/s00216-006-0824-4
PubMed ID:
17143597
Type:
Article
Language:
en
ISSN:
1618-2642
Appears in Collections:
collections of the research group microbial communication (KOM)

Full metadata record

DC FieldValue Language
dc.contributor.authorVilchez, Ramiro-
dc.contributor.authorLemme, André-
dc.contributor.authorThiel, Verena-
dc.contributor.authorSchulz, Stefan-
dc.contributor.authorSztajer, Helena-
dc.contributor.authorWagner-Döbler, Irene-
dc.date.accessioned2007-07-30T10:21:30Z-
dc.date.available2007-07-30T10:21:30Z-
dc.date.issued2007-01-01-
dc.identifier.citationAnal Bioanal Chem 2007, 387(2):489-96en
dc.identifier.issn1618-2642-
dc.identifier.pmid17143597-
dc.identifier.doi10.1007/s00216-006-0824-4-
dc.identifier.urihttp://hdl.handle.net/10033/12985-
dc.description.abstractAutoinducer-2 (furanosyl borate diester) is a biologically active compound whose role as a universal bacterial signalling molecule is currently under intense investigation. Because of its instability and the low concentrations of it found in biological samples, its detection relies at present on a bioassay that measures the difference in the timing of the luminescence of the Vibrio harveyi BB170 sensor strain with and without externally added AI-2. Here we systematically investigated which parameters affected the fold induction values of luminescence obtained in the bioassay and developed a modified protocol. Our experiments showed that growth and luminescence of V. harveyi BB170 are strongly influenced by trace elements. In particular, addition of Fe(3+) within a certain concentration range to the growth medium of the preinoculum culture improved the reproducibility and reduced the variance of the bioassay. In contrast, trace elements and vitamins introduced directly into the bioassay caused inhibitory effects. The initial density and luminescence of the sensor strain are very important and the values required for these parameters were defined. Borate interferes with the detection of AI-2 by giving false positive results. The response of V. harveyi BB170 to chemically synthesized AI-2 in the bioassay is nonlinear except over a very small concentration range; it is maximum over three orders of magnitude and shows inhibition above 35 microM. Based on the modified protocol, we were able to detect AI-2 in the absence of inhibitors with maximum fold induction values for the positive control (chemically synthesized AI-2) of >120 with a standard deviation of approximately 30% in a reliable and reproducible way.en
dc.format.extent221061 bytes-
dc.format.mimetypeapplication/pdf-
dc.language.isoenen
dc.titleAnalysing traces of autoinducer-2 requires standardization of the Vibrio harveyi bioassay.en
dc.typeArticleen
dc.format.digYES-

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