Isolation and Characterization of Lactic Acid Bacteria from Winery Wastes
Maria Rosa Morales, Maria Jose Rodriguez-Vaquero,* and Fabiana Maria Saguir *CONICET-UNT, Ayacucho 471, 4000, Argentina (mariajo@fbqf.unt.edu.ar)
Wineries create large volumes of byproduct streams and waste,
including grape pomace, wine lees, and grape stalk, which should
be valorized or treated before disposal to prevent environmental
pollution. Grape pomace is the solid residue left after juice
extraction during winemaking and includes grape skins and seeds.
Wine lees are the residue that forms at the bottom of recepticals
containing wine, after fermentation or during storage. The aim of
this work was to isolate and characterize the lactic acid
bacteria (LAB) present in both pomace and lees to find LAB with
optimal properties to be included in fermented food. Malbec grape
pomace and wine lees samples were centrifuged and resuspended in
MRS medium enriched with 5 g/L fructose and 3 g/L L-malic acid
and adjusted to pH 5.0. After incubation at 30°C for 72 hr,
samples were plated on MRS agar supplemented with 1%
cycloheximide (MRS-C). All plates were incubated aerobically at
30°C for 72 hr. Twenty colonies of each sample were selected and
preliminarily identified by cell morphology, Gram stain, catalase
and dextran production, and growth under different conditions,
such as NaCl concentrations (2.5, 5.0, or 7.5%),
pH (4.0, 7.0, or 9.0), and temperature (15, 30, or 45°C). Glucose
consumption and D- or L-lactic acid isomer production were also
determined. Of 40 isolated bacteria, 28 were LAB and produced
both isomers of lactic acid from glucose. All LAB isolated from
grape pomace and most from lees isolated were homofermentative;
only six were heterofermentative. Our results showed the presence
of LAB in winery wastes generally had high lactic acid production
and optimal properties for inclusion in wine or other beverages,
but further studies must be conducted.
Funding Support: PICT 2015-1508 – Prestamo Bid PIP-CONICET CIUNT