Architecture and physicochemical characterization of Bacillus biofilm as a potential enzyme immobilization factory

Biocatalysis for industrial application is based on the use of enzymes to perform complex transformations. However, these systems have some disadvantage related to the costs of the biocatalyst. In this work, an alternative strategy for producing green immobilized biocatalysts based on biofilm was developed.A study of the rheological behavior of the biofilm from Bacillus sp. Mcn4, as well as the determination of its composition, was carried out. The dynamic rheological measurements, viscosity (G") and elasticity (G') module, showed that the biofilm presents appreciable elastic components, which is a recognized property for enzymes immobilization. After the partial purification, the exopolysaccharidewas identified as a levan with a non-Newtonian behavior. Extracellular DNA with fragments between 10,000 and 1000bp was detected also in the biofilm, and amyloid protein in the extracellular matrix using a fluorescence technique was identified. Bacillus sp. Mcn4 biofilms were developed on different surfaces, being the most stable those developed on hydrophilic supports. The biofilm showed lipase activity suggesting the presence of constitutive lipases entrapped into the biofilm. Indeed, two enzymes with lipase activity were identified in native PAGE. These were used as biocatalysts, whose reuse showed a residual lipase activity after more than one cycle of catalysis. The components identified in the biofilm could be the main contributors of the rheological characteristic of this material, giving an exceptional environment to the lipase enzyme. Based on these findings, the current study proposes green and natural biopolymers matrix as support for the enzyme immobilization for industrial applications.