β‑(1 → 3)-Glucanolytic Yeasts from Brazilian Grape Microbiota: Production and Characterization of β‑Glucanolytic Enzymes by Aureobasidium pullulans 1WA1 Cultivated on Fungal Mycelium

• Published in: Journal of agricultural and food chemistry Vol. 63; no. 1; pp. 269 - 278
• Main Authors: Bauermeister, Anelize, Amador, Ismael R, Pretti, Carla P, Giese, Ellen C, Oliveira, André L. M, Alves da Cunha, Mário A, Rezende, Maria Inês, Dekker, Robert F. H, Barbosa, Aneli M
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 14.01.2015
• Subjects: Ascomycota - classification; Ascomycota - enzymology; Ascomycota - genetics; Ascomycota - isolation & purification; beta-Glucans - metabolism; Brazil; Fungal Proteins - chemistry; Fungal Proteins - genetics; Fungal Proteins - metabolism; Glucans - chemistry; Glucans - metabolism; Glycoside Hydrolases - chemistry; Glycoside Hydrolases - genetics; Glycoside Hydrolases - metabolism; Hydrolysis; Kinetics; Microbiota; Mycelium - chemistry; Mycelium - enzymology; Mycelium - genetics; Substrate Specificity; Vitis - microbiology; Yeasts - classification; Yeasts - genetics; Yeasts - isolation & purification; Brazil; Aureobasidium pullulans; Botryosphaeria rhodina; β-(1 → 3)-glucanases; β-d-glucans; botryosphaeran; grape microbiota yeasts; β-glucosidases
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/jf504333h

A total of 95 yeast strains were isolated from the microbiota of different grapes collected at vineyards in southern Brazil. The yeasts were screened for β-(1 → 3)-glucanases using a newly developed zymogram method that relies upon the appearance of clearance zones around growing colonies cultured on agar–botryosphaeran medium and also by submerged fermentation on nutrient medium containing botryosphaeran, a (1 → 3),(1 → 6)-β-d-glucan. Among 14 β-(1 → 3)-glucanase-positive yeasts identified, four strains produced the highest β-glucanolytic activities and were evaluated for enzyme production on cellobiose, botryosphaeran, and mycelial biomass from Botryosphaeria rhodina (MAMB-05). Yeast strain 1WA1 produced the highest β-(1 → 3)-glucanase and β-glucosidase activities and was identified by molecular characterization as Aureobasidium pullulans. The physicochemical properties of the crude β-glucanolytic enzyme preparation were characterized, and the preparation was used to hydrolyze several β-d-glucans (laminarin, botryosphaeran, lasiodiplodan, pustulan, and curdlan). The production and physicochemical properties of the β-glucanolytic preparation enable its potential applications in wine enology and production of prebiotics through hydrolysis of β-d-glucans.