Enhancement of beta-1,3 Glucanase Production from Penicillium oxalicum T3.3
Abstract
β-1,3 glucanases are semi-constitutive hydrolytic enzymes that can degrade glucan molecules embedded in the cell wall components of cereals and some species of fungi resulted in production of D-glucose. This enzyme has a great potential and interest in biotechnology, agricultural and also industrial field. However, there is little reports on the production of β-1,3 glucanase by Penicillium oxalicum. Therefore, the cultural conditions which stimulate in vitro production of β-1,3 glucanase enzyme by P. oxalicum T3.3 and characterization of β-1,3 glucanase enzyme activity were determined.Various parameters such as different types of carbon and nitrogen sources, initial pH medium, agitation speed and surfactants were investigated. The optimization was carried out by varying and optimizing one variable at a time. The highest production of β-1,3 glucanase activity of 84.73 U/mL was obtained using seaweed Undaria pinnatifida as substrate at concentration of 1% (w/v), peptone and yeast extract as nitrogen source at 0.3% and 0.2% respectively, initial medium pH 5, agitation speed at 200 rpm and with addition of sodium dodecyl sulfate as surfactant. Under these conditions, β-1,3 glucanase activity increased by 38.6%. Enzyme characterization was also performed which indicated that this enzyme is thermostable and showed optimum activity at 50°C, pH 5 and can retained its activity around 80% up to 4 hat this condition.The optimization of β-1,3 glucanase production by P.oxalicum required adjustment of different types of carbon and nitrogen sources, initial pH medium, agitation speed and surfactants. This enzyme characterization has revealed its great potential towards detergent, beer and food fermentation industries whose manufacturing conditions are largely acidic.
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Introduction
β-1,3-glucanase is an important enzyme in the industrial and agricultural processing field. β-1,3 glucanase enzymes have received attention in many fields of science and biotechnology because many cultures of microorganisms widely used in industry produce β-1,3 glucanase (Kulminskaya etal., 2001). The resistance of this enzyme to denaturation by high temperature and pH extremes makes it particularly essential in various functions (Beshay et al., 2011). β-1,3-glucanases have been reported to be produced by a variety of organisms such as bacteria, fungi, and higher plants and many of them have been purified and characterized (Martin et al., 2007). β-1,3 glucanase can be used as biocontrol agent againstplant-pathogenic fungi and act as hydrolyticenzyme for the destruction of cell wall of fungal and structure of β-1,3-glucan (Beshay et al., 2003). Besides, for industrial purpose they are used commercially in combination with other enzymes in the production of beer and in the brewery, for barley-beta glucan degradation for animal feed. Furthermore, they can be used as effective additives in laundry detergents. They can also be used for saccharification of agricultural and industrial wastes to provide glucose syrups for animal use (Doughari and Hamuel 2011). Penicillium sp has also been reported to produce hydrolytic enzymes including chitinase and β-glucanase which are involved in the degrading of fungal cell wall (Chen et al., 2012; Patil et al., 2013). In another study, it was suggested that P. oxalicum secretes chitinase and β-glucanases to degrade and penetrate into the conidiophores and spores of Nigrospora oryzae (Sempere and Santamarina, 2008).
To the best of our knowledge there is little reports on the production of β-1,3 glucanases by P. oxalicum. Thus, a study on the optimization of β-1,3 glucanases production using this fungus is important to be carried out in order to enhance β-1,3 glucanases production. The most important aspects to decrease the production cost are optimization of media and process conditions (Goshal et al., 2011). In this study, optimization of β-1,3 glucanases production was done by using the conventional method, which involved varying one variable at a time while the other variables were kept constant. Therefore, the objectives of this study were to determine the cultural conditions which stimulate the in vitro production of β-1,3 glucanase enzyme by P. oxalicum T3.3 and to characterize the β-1,3 glucanase enzyme activity.
Conclusion
The optimization of β-1,3 glucanase production by P.oxalicum required adjustment of different types of carbon and nitrogen sources, initial pH medium, agitation speed and surfactants. This enzyme characterization has revealed its great potential towards detergent, beer and food fermentation industries whose manufacturing conditions are largely acidic.
