Screening for glycosidase activities of lactic acid bacteria as a biotechnological tool in oenology.
The aim of this study was to evaluate the ability from a number of lactic acid bacteria isolated from different sources to produce glycosidase enzymes. Representative isolates (225) from clusters obtained after genotyping, using randomly amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) analysis, of 1,464 isolates, were screened for β-D-glucosidase activity. Thirty-five of them were selected for subsequent analysis. These strains were able to hydrolyze α-D-glucopyranoside, β-D-xylopyranoside and α-L-arabinofuranoside although β-D-glucosidase activity was the predominant activity for 22 of the selected strains. Only some of them did so with α-L-rhamnopyranoside. All of these were from wine samples and were identified as belonging to the Oenococcus oeni species using Amplification and Restriction Analysis of 16S-rRNA gene (16S-ARDRA). When the influence of pH, temperature and ethanol or sugars content on β-D-glucosidase activity was assayed, a strain-dependent response was observed. The β-D-glucosidase activity occurred in both whole and sonicated cells but not in the supernatants from cultures or obtained after cell sonication. Strains 10, 17, 21, and 23 retained the most β-D-glucosidase activity when they were assayed at the conditions of temperature, pH, ethanol and sugar content used in winemaking. These results suggest that these strains could be used as a source of glycosidase enzymes for use in winemaking.
Department of Analytical Chemistry and Food Technology, Environmental Sciences and Biochemistry Faculty, University of Castilla-La Mancha, Avda. Carlos III, s/n, 45071, Toledo, Spain.
Fátima
F
Pérez-Martín
Susana
S
Seseña
Pedro Miguel
PM
Izquierdo
Raúl
R
Martín
María Llanos
ML
Palop
D001709:Biotechnology / Q000379:methods*; D004269:DNA, Bacterial / Q000235:genetics; D004791:Enzyme Inhibitors / Q000378:metabolism; D000431:Ethanol / Q000378:metabolism; D020459:Genes, rRNA; D006026:Glycoside Hydrolases / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D056546:Lactobacillales / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D008403:Mass Screening / Q000379:methods*; D058889:Molecular Typing; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013696:Temperature; D014920:Wine / Q000382:microbiology*
D004269:DNA, Bacterial; D004791:Enzyme Inhibitors; D012336:RNA, Ribosomal, 16S; D000431:Ethanol; D006026:Glycoside Hydrolases