Enhancing Laccase Production by Trametes hirsuta GMA-01 Using Response Surface Methodology and Orange Waste: A Novel Breakthrough in Sugarcane Bagasse Saccharification and Synthetic Dye Decolorization

Author(s)

Ortolan, Guilherme Guimaraes ; Contato, Alex Graca ; Aranha, Guilherme Mauro ; Salgado, Jose Carlos Santos ; Alnoch, Robson Carlos ; Polizeli, Maria de Lourdes Teixeira de Moraes

• Document type: Journal article
• Source: REACTIONS; v. 5, n. 3, p. 16-pg., 2024-09-01.

Abstract

Trametes hirsuta GMA-01 was cultivated in a culture medium supplemented with orange waste, starch, wheat bran, yeast extract, and salts. The fungus produced several holoenzymes, but the laccase levels were surprisingly high. Given the highlighted applicability of laccases in various biotechnological areas with minimal environmental impact, we provided a strategy to increase its production using response surface methodology. The immobilization of laccase into ionic supports (CM-cellulose, DEAE-agarose, DEAE-cellulose, DEAE-Sephacel, MANAE-agarose, MANAE-cellulose, and PEI-agarose) was found to be efficient and recuperative, showcasing the technical prowess of research. The crude extract laccase (CE) and CM-cellulose-immobilized crude extract (ICE) showed optimum activity in acidic conditions (pH 3.0) and at 70 degrees C for the CE and 60 degrees C for the ICE. The ICE significantly increased thermostability at 60 degrees C for the crude extract, which retained 21.6% residual activity after 240 min. The CE and ICE were successfully applied to sugarcane bagasse hydrolysis, showing 13.83 +/- 0.02 mu mol mL(-1) reducing sugars after 48 h. Furthermore, the CE was tested for dye decolorization, achieving 96.6%, 71.9%, and 70.8% decolorization for bromocresol green, bromophenol blue, and orcein, respectively (0.05% (w/v) concentration). The properties and versatility of T. hirsuta GMA-01 laccase in different biotechnological purposes are interesting and notable, opening several potential applications and providing valuable insights into the future of biotechnological development. (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: 14/50884-5 - INCT 2014: National Institute of Science and Technology of Bioethanol
• Grantee: Marcos Silveira Buckeridge
• Support Opportunities: Research Projects - Thematic Grants
• 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: 19/21989-7 - Bioprospection of low molecular weight fungal enzymes involved in plant cell wall degradation
• Grantee: José Carlos dos Santos Salgado
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• 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: 23/09627-8 - Formulation of tailor-made enzymatic consortiums for the degradation of the cell wall of sugarcane varieties
• Grantee: Robson Carlos Alnoch
• Support Opportunities: Scholarships in Brazil - Support Program for Fixating Young Doctors
• FAPESP's process: 23/01547-5 - Formulation of tailor-made enzymatic consortiums for the degradation of the cell wall of sugarcane varieties
• Grantee: Maria de Lourdes Teixeira de Moraes Polizeli
• Support Opportunities: Regular Research Grants