Cellulase production by Aspergillus niger using urban lignocellulosic waste as substrate: Evaluation of different cultivation strategies

Abstract

Cellulases are used in various industries, acting efficiently and sustainably in the degradation of cellulose contained in different raw materials and recovering high value products. It is the third largest group of enzymes consumed industrially, as they are required in processes linked to the food, biofuel, textile, cleaning products, among others. However, the main disadvantage in the use of commercial cellulases is the high cost. In this context, the objective of this work was to determine conditions for obtaining more efficient and economical cellulases. For this, the efficiency in obtaining the extracellular cellulases endoglucanase (CMCase) and exoglucanase (FPase) by a fungus Aspergillus niger was investigated using an urban lignocellulosic waste as substrate characterized by tree leaves collected from squares and avenues in urban areas. As urban lignocellulosic waste is an innovative raw material, its chemical composition was determined. This substrate contains 20.36% cellulose and induced the production of cellulases in all fermentation methods, proving to be a promising and sustainable source. The influence of the nutrient medium on CMCase and FPase activities was evaluated for three different sequential fermentation (SF) configurations. Medium 2 provided an increase of up to 100 U/L of CMCase and FPase in relation to medium 1. The interactive effect of pH and moisture content on CMCase e FPase production under SF was studied in a central composite design (CCD). Also, different fermentation methods (solid state, submerged and sequential) were evaluated. The use of SF increased the enzymatic activities of both cellulases by 140% compared to other conventional methods and also stood out in the production of proteins (270.05 μg/mL) and reducing sugars (1.19 mg/mL). The desirability function determined the optimal activities of CMCase and FPase as 413.49 U/L and 230.68 U/L, respectively, obtained from the optimal variables of pH 5.5 and 75% moisture content under SF. The effect of pH and moisture content on the activity of each cellulase was analyzed using the Pareto chart and response surface methodology (RSM). These results revealed favorable strategies for cellulase production, such as the use of urban lignocellulosic waste, SF and ideal operational conditions.