Application of a modified drop-collapse technique for surfactant quantitation and screening of biosurfactant-producing microorganisms

Adria A. Bodour, Raina Margaret Maier

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259 Citations (Scopus)

Abstract

A drop-collapse method has been refined for use as both a qualitative assay to screen for surfactant-producing microbes, and as a quantitative assay to determine surfactant concentration. The assay is rapid, easy to perform, reproducible and requires little specialized equipment. The assay is performed in a 96-microwell plate, where each well is thinly coated with oil. A 5 μL sample droplet is added to the center of a well and observed after 1 min. The droplet will either bead up, spread out slightly or collapse, depending on the amount of surfactant in the sample. The basis for this method is the type of oil used to coat each well. In the qualitative method, each well is coated with 1.8 μL of Pennzoil® and either the drop collapses, indicating the presence of surfactant (a positive result), or the drop remains beaded, indicating the absence of surfactant (a negative response). In the quantitative method, each well is coated with 2 μL of mineral oil, and a dissecting microscope is used to measure the diameter of the droplet at 1 min. Results with both a test biosurfactant (rhamnolipid) and a test synthetic surfactant (sodium dodecyl sulfate) indicate a direct linear correlation between droplet diameter and surfactant concentration. The drop-collapse method has several advantages over commonly used methods that measure surface tension, such as the du Nouy ring method; a smaller volume is required (5 μL vs. 20 mL), the effective range of measurement is greater and it does not require specialized equipment. Copyright (C) 1998 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)273-280
Number of pages8
JournalJournal of Microbiological Methods
Volume32
Issue number3
DOIs
StatePublished - May 1 1998

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Surface-Active Agents
Oils
Mineral Oil
Equipment and Supplies
Surface Tension
Sodium Dodecyl Sulfate

Keywords

  • Biosurfactant
  • Drop-collapse test
  • Screening assay
  • Surfactant

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Microbiology

Cite this

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abstract = "A drop-collapse method has been refined for use as both a qualitative assay to screen for surfactant-producing microbes, and as a quantitative assay to determine surfactant concentration. The assay is rapid, easy to perform, reproducible and requires little specialized equipment. The assay is performed in a 96-microwell plate, where each well is thinly coated with oil. A 5 μL sample droplet is added to the center of a well and observed after 1 min. The droplet will either bead up, spread out slightly or collapse, depending on the amount of surfactant in the sample. The basis for this method is the type of oil used to coat each well. In the qualitative method, each well is coated with 1.8 μL of Pennzoil{\circledR} and either the drop collapses, indicating the presence of surfactant (a positive result), or the drop remains beaded, indicating the absence of surfactant (a negative response). In the quantitative method, each well is coated with 2 μL of mineral oil, and a dissecting microscope is used to measure the diameter of the droplet at 1 min. Results with both a test biosurfactant (rhamnolipid) and a test synthetic surfactant (sodium dodecyl sulfate) indicate a direct linear correlation between droplet diameter and surfactant concentration. The drop-collapse method has several advantages over commonly used methods that measure surface tension, such as the du Nouy ring method; a smaller volume is required (5 μL vs. 20 mL), the effective range of measurement is greater and it does not require specialized equipment. Copyright (C) 1998 Elsevier Science B.V.",
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