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Invited Review: Methods for the Screening, Isolation, and Characterization of Exopolysaccharides Produced by Lactic Acid Bacteria

Instituto de Productos Lácteos de Asturias, CSIC, Carretera de Infiesto s/n, 33300 Villaviciosa, Asturias, Spain

Corresponding author: Patricia Ruas-Madiedo; e-mail: ruas-madiedo{at}ipla.csic.es.

The ability to produce exopolysaccharides (EPS) is widespread among lactic acid bacteria (LAB), although the physiological role of these molecules has not been clearly established yet. Some EPS confer on LAB a "ropy" character that can be detected in cultures that form long strands when extended with an inoculation loop. When EPS are produced in situ during milk fermentation they can act as natural biothickeners, giving the product a suitable consistency, improving viscosity, and reducing syneresis. In addition, some of these EPS may have beneficial effects on human health. The increasing demand by consumers of novel dairy products requires a better understanding of the effect of EPS on existing products and, at the same time, the search for new EPS-producing strains with desirable properties. The use of genetically modified organisms capable of producing high levels of EPS or newly designed biopolymers is still very limited. Therefore, exploration of the biodiversity of wild LAB strains from natural ecological environments is currently the most suitable approach to search for the desired EPS-phenotype. The screening of ropy strains and the isolation and characterization of EPS responsible for this characteristic have led to the application over the past years of a wide variety of techniques. This review summarizes the available information on methods and procedures used for research on this topic. The information provided deals with methods for screening of EPS-producing LAB, detection of the ropy phenotype, and the physicochemical and structural characterization of these molecules, including parameters related to their viscosifying properties. To our knowledge, this is the first compilation of methods available for the study of EPS produced by LAB.

Key Words: exopolysaccharide • screening • isolation • characterization

Abbreviation key: CSLM = confocal scanning laser microscopy, ESM = EPS selection media, EPS = exopo-lysaccharides, GCMS = gas chromatography/mass spectrometry, HePS = heteropolysaccharides, HoPS = homopolysaccharides, HPAEC-PAD = high-performance anion-exchange chromatography pulse ampero-metric detection, LAB = lactic acid bacteria, MRS = De Man, Rogosa, Sharpe, NIRS = near infrared spectroscopy, Rg = radius of gyration, RI = refractive index.

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