Thermothelomyces thermophilus cultivated with residues from the fruit pulp industry: enzyme immobilization on ionic supports of a crude cocktail with enhanced production of lichenase

Author(s)

Contato, Alex Graca ; Vici, Ana Claudia ; Pinheiro, Vanessa Elisa ; de Oliveira, Tassio Brito ; Ortolan, Guilherme Guimaraes ; de Freitas, Emanuelle Neiverth ; Buckeridge, Marcos Silveira ; Polizeli, Maria de Lourdes Teixeira de Moraes

• Document type: Journal article
• Source: Folia Microbiologica; v. 70, n. 3, p. 11-pg., 2024-10-23.

Abstract

beta-Glucans comprise a group of beta-d-glucose polysaccharides (glucans) that occur naturally in the cell walls of bacteria, fungi, and cereals. Its degradation is catalyzed by beta-glucanases, enzymes that catalyze the breakdown of beta-glucan into cello-oligosaccharides and glucose. These enzymes are classified as endo-glucanases, exo-glucanases, and glucosidases according to their mechanism of action, being the lichenases (beta-1,3;1,4-glucanases, EC 3.2.1.73) one of them. Hence, we aimed to enhance lichenase production by Thermothelomyces thermophilus through the application of response surface methodology, using tamarind (Tamarindus indica) and jatoba (Hymenaea courbaril) seeds as carbon sources. The crude extract was immobilized, with a focus on improving lichenase activity, using various ionic supports, including MANAE (monoamine-N-aminoethyl), DEAE (diethylaminoethyl)-cellulose, CM (carboxymethyl)-cellulose, and PEI (polyethyleneimine)-agarose. Regarding lichenase, the optimal conditions yielding the highest activity were determined as 1.5% tamarind seeds, cultivation at 50 degrees C under static conditions for 72 h. Moreover, transitioning from Erlenmeyer flasks to a bioreactor proved pivotal, resulting in a 2.21-fold increase in activity. Biochemical characterization revealed an optimum temperature of 50 degrees C and pH of 6.5. However, sustained stability at varying pH and temperature levels was challenging, underscoring the necessity of immobilizing lichenase on ionic supports. Notably, CM-cellulose emerged as the most effective immobilization medium, exhibiting an activity of 1.01 U/g of the derivative (enzyme plus support), marking a substantial enhancement. This study marks the first lichenase immobilization on these chemical supports in existing literature. (AU)

• FAPESP's process: 17/25862-6 - Structural studies and functional properties of xyloglucanases and lichenases: applications on hydrolysis of lignocellulosic biomass
• Grantee: Alex Graça Contato
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 18/07522-6 - Development of eco-friendly technologies for the hydrolysis of biomass and residues of cellulose pulp and paper industry
• Grantee: Maria de Lourdes Teixeira de Moraes Polizeli
• Support Opportunities: Regular Research Grants
• FAPESP's process: 22/06275-0 - Development of techniques for immobilization, purification, and characterization of Trametes hirsuta laccases aiming at biotechnological applications
• Grantee: Guilherme Guimarães Ortolan
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 21/07066-3 - Molecular studies of a Trichoderma longibrachiatum xyloglucanase: cloning, heterologous expression, purification and characterization
• Grantee: Alex Graça Contato
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• FAPESP's process: 17/23989-9 - Lignocellulosic biomass of Panicum maximum Jacq. forage grass produced in simulated future climate conditions: potential source of fermentable sugars for bioenergy
• Grantee: Emanuelle Neiverth de Freitas
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 17/09000-4 - Effects of temperature and water stress, in climate change scenario, on the fungal community in tropical soil and selection of nitrous oxide producing strains
• Grantee: Tássio Brito de Oliveira
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 14/50884-5 - INCT 2014: National Institute of Science and Technology of Bioethanol
• Grantee: Marcos Silveira Buckeridge
• Support Opportunities: Research Projects - Thematic Grants