XTT assay for detection of bacterial metabolic activity in water-based polyester polyurethane.

Autor: Magaña-Montiel N; Departamento de Microbiología Molecular, Instituto de Biotecnología, Cuernavaca, Morelos, México., Muriel-Millán LF; Departamento de Microbiología Molecular, Instituto de Biotecnología, Cuernavaca, Morelos, México., Pardo-López L; Departamento de Microbiología Molecular, Instituto de Biotecnología, Cuernavaca, Morelos, México.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2024 Jun 06; Vol. 19 (6), pp. e0303210. Date of Electronic Publication: 2024 Jun 06 (Print Publication: 2024).
DOI: 10.1371/journal.pone.0303210
Abstrakt: Cellular metabolic activity can be detected by tetrazolium-based colorimetric assays, which rely on dehydrogenase enzymes from living cells to reduce tetrazolium compounds into colored formazan products. Although these methods have been used in different fields of microbiology, their application to the detection of bacteria with plastic-degrading activity has not been well documented. Here, we report a microplate-adapted method for the detection of bacteria metabolically active on the commercial polyester polyurethane (PU) Impranil®DLN using the tetrazolium salt 2,3-bis [2-methyloxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide (XTT). Bacterial cells that are active on PU reduce XTT to a water-soluble orange dye, which can be quantitatively measured using a microplate reader. We used the Pseudomonas putida KT2440 strain as a study model. Its metabolic activity on Impranil detected by our novel method was further verified by Fourier-transform infrared spectroscopy (FTIR) analyses. Measurements of the absorbance of reduced XTT at 470 nm in microplate wells were not affected by the colloidal properties of Impranil or cell density. In summary, we provide here an easy and high-throughput method for screening bacteria active on PU that can be adapted to other plastic substrates.
Competing Interests: The authors have declared that no competing interests exist.
(Copyright: © 2024 Magaña-Montiel et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
Databáze: MEDLINE
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