Background

The cellulolytic enzyme complex has many important biotechnological applications such as beverage, textile, food, paper and cellulose industries, as well as in degra-dation of lignocellulosic for ethanol biofuel production [1]. According to the enzymatic activity, cellulolytic complex is subdivided in three classes: endoglucanases, exoglucanases and b-D-glycosidases. Endoglucanases (endo-b-1,4-glucanase, EC 3.2.1.4) are responsible to initiate cleavage, hydrolyzing randomly internal regions from the amorphous cellulose fiber structures, releasing oligosaccharides from different grades of polymerization. These will be hydrolyzed by exoglucanases releasing cellobiose followed by b-D-gly-cosidades being hydrolyzed to glucose [1]. Penicillium citrinum has worldwide occurrence. Com-monly in the soil, this specie is described as good xyla-nase and cellulase producer [2,3]. This work describes liquid chromatography to separate endoglucanases of Penicillium citrinum supernatant from submerse fermentation process. Methods Suspension of 1 mL P. citrinum spores (6.4 × 106) was inoculated in 200 mL submerse fermentation medium [4] containing CMC as carbon source for 10 days/28°C/ 150 rpm. Every 24 hours was sampled 6 mL aliquots, centrifuged and right after the supernatant storage at 4° C. A pool was made of collected points from 72 to 240 hours. It was concentrated in SpeedVac (Thermo) for final volume of 1.5 mL, filtrate (0.45 µm) and injected in chromatograph column Superdex 75 10/300 GL (GE Healthcare) previously equilibrated with sodium citrate buffer at 50 mM pH 4.8. Fractions of 1.5 mL was col-lected in a flux of 0.6 mL/min in 1.5 column volume