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Isolation of amylolytic,xylanolytic, and cellulolytic microorganisms extracted from the gut of the termite Reticulitermes santonensis by means of a micro-aerobic atmosphere
Authors:Cédric Tarayre  Alison Brognaux  Julien Bauwens  Catherine Brasseur  Christel Mattéotti  Catherine Millet  Jacqueline Destain  Micheline Vandenbol  Daniel Portetelle  Edwin De Pauw  Haubruge Eric  Frédéric Francis  Philippe Thonart
Affiliation:1. Bat. G1 Bio-Industries, Passage des Déportés 2, 5030, Gembloux, Belgium
2. Bat. G1 Entomologie Fonctionnelle et évolutive, Passage des Déportés 2, 5030, Gembloux, Belgium
3. Bat. B6C, Laboratoire de Spectrométrie de Masse (L.S.M.), Allée de la Chimie 3, 4000, Liège 1, Belgium
4. Bat. G1 Unité de Microbiologie et Génomique, Avenue Maréchal Juin 6, 5030, Gembloux, Belgium
5. Bat. G1 Vice-Recteur de Gembloux Agro-Bio Tech, Passage des Déportés 2, 5030, Gembloux, Belgium
6. Bat. B40 Biochimie et microbiologie industrielles, Boulevard du Rectorat 29, 4000, Liège 1, Belgium
Abstract:The aim of this work was to isolate enzyme-producing microorganisms from the tract of the termite Reticulitermes santonensis. The microorganisms were extracted from the guts and anaerobic (CO2 or CO2/H2) and micro-aerobic atmospheres were used to stimulate growth. Three different strategies were tried out. First, the sample was spread on Petri dishes containing solid media with carboxymethylcellulose, microcrystalline cellulose or cellobiose. This technique allowed us to isolate two bacteria: Streptomyces sp. strain ABGxAviA1 and Pseudomonas sp. strain ABGxCellA. The second strategy consisted in inoculating a specific liquid medium containing carboxymethylcellulose, microcrystalline cellulose, or cellobiose. The samples were then spread on Petri dishes with the same specific medium containing carboxymethylcellulose, microcrystalline cellulose, or cellobiose. This led to the isolation of the mold Aspergillus sp. strain ABGxAviA2. Finally, the third strategy consisted in heating the first culture and spreading samples on agar plates containing rich medium. This led to the isolation of the bacterium Bacillus subtilis strain ABGx. All those steps were achieved in controlled atmospheres. The four enzyme-producing strains which were isolated were obtained by using a micro-aerobic atmosphere. Later, enzymatic assays were performed on the four strains. Streptomyces sp. strain ABGxAviA1 was found to produce only amylase, while Pseudomonas sp. strain ABGxCellA was found to produce β-glucosidase as well. Aspergillus sp. strain ABGxAviA2 showed β-glucosidase, amylase, cellulase, and xylanase activities. Finally, B. subtilis strain ABGx produced xylanase and amylase.
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