Calcium alginate-immobilized β-glucosidase from Moniliophthora perniciosa : characterization and sugarcane bagasse hydrolysis

Abstract

AbstractThe utilization of lignocellulosic materials for second-generation ethanol production via enzymatic catalysts is primarily hindered by enzyme cost. Enzymatic immobilization emerges as a viable solution, enabling enzyme reuse. This study investigated the immobilization of an enzymatic extract obtained from Moniliophthora perniciosa fermentation in calcium alginate spheres using the direct trapping method. Initial tests assessed β-glucosidase activity, showing that a higher concentration of calcium chloride (1 M) alongside larger diameter spheres yielded improved results. The immobilized enzyme was reused for up to 17 cycles without significant loss of activity. The percentage of reducing sugars after 48-h hydrolysis with the supplemented enzymatic extract was 226%, doubling the value achieved with only the free enzymatic extract. The immobilized enzyme retained 50% of its initial activity after 1 h at 80 °C, demonstrating higher activity at pH 6 and 60 °C. These findings suggest that this immobilization technique is simple, economically viable, and effective for the hydrolysis of pretreated sugarcane bagasse.HIGHLIGHTSSuccessful immobilization of M. perniciosa enzymatic extract achieved through direct entrapment in calcium alginate.Immobilized β-glucosidase demonstrates sustained activity over 16 reuse cycles, showcasing the potential for cost-effective bioconversion processes.Enhanced hydrolysis of sugarcane bagasse observed with immobilized enzymatic extract, indicating a promising approach for improved biomass utilization.Keywords: β-Glucosidase immobilizationmoniliophthora perniciosacalcium alginate spheressugarcane bagasse hydrolysisenzymatic bioconversionlignocellulosic ethanolreducing sugars AcknowledgmentsWe thank the Biotechnology Graduate Program of State University of Feira de Santana (UEFS/FIOCRUZ), the Coordenação de Aperfeiçoamento Pessoal de Nível Superior (CAPES) for a doctoral scholarship (88882.447813/2019-01), the Bahia State Research Support Foundation (FAPESB), and the National Council for Scientific and Technological Development (CNPq).Authors’ contributionsAll authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by Larissa E. S. Almeida and Sandra A. Assis. The first draft of the manuscript was written by Larissa E. S. Almeida, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.Disclosure statementNo potential conflict of interest was reported by the author(s).Additional informationFundingWe thank the Biotechnology Graduate Program of State University of Feira de Santana (UEFS/FIOCRUZ), the Coordenação de Aperfeiçoamento Pessoal de Nível Superior (CAPES) for a doctoral scholarship (88882.447813/2019-01), the Bahia State Research Support Foundation (FAPESB), and the National Council for Scientific and Technological Development (CNPq).