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Characterization of the cellulase-secretome produced by the Antarctic bacterium Flavobacterium sp. AUG42.

Microbiological research (2019-06-11)
Lorena M Herrera, Victoria Braña, Laura Franco Fraguas, Susana Castro-Sowinski
ABSTRACT

Flavobacterium sp. AUG42 is a cellulase-producing bacterium isolated from the Antarctic oligochaete Grania sp. (Annelida). In this work, we report that AUG42 produces a glycoside hydrolase cocktail with CMCase, PASCase and cellobiase activities (optimum pHs and temperatures ranging from 5.5 to 6.5 and 40 to 50 °C, respectively). The time-course analyses of the bacterial growth and cellulase production showed that the cocktail has maximal activity at the stationary phase when growing at 16 °C with filter paper as a cellulosic carbon source, among the tested substrates. The analyses of the CAZome and the identification of secreted proteins by shotgun Mass Spectrometry analysis showed that five glycoside hydrolyses are present in the bacterial secretome, which probably cooperate in the degradation of the cellulosic substrates. Two of these glycoside hydrolyses may harbor putative carbohydrate binding modules, both with a cleft-like active site. The cellulolytic cocktail was assayed in saccharification experiments using carboxymethylcellulose as a substrate and results showed the release of glucose (a fermentable sugar) and other reducing-sugars, after 24 h incubation. The ecological relevance of producing cellulases in the Antarctic environment, as well as their potential use in the bio-refinery industry, are discussed.

MATERIALS
Product Number
Brand
Product Description

Millipore
D-(+)-Cellobiose, ≥99.0%, suitable for microbiology
Sigma-Aldrich
D-(+)-Cellobiose, ≥98%
Sigma-Aldrich
Carboxymethylcellulose sodium salt, low viscosity
Sigma-Aldrich
Cellulose, microcrystalline, powder
Sigma-Aldrich
Avicel® PH-101, tested according to Ph. Eur.