Co-immobilization of multiple enzymes on ferromagnetic nanoparticles for the depolymerization of xyloglucan

Author(s)

Soares, Jessica M. ; Carneiro, Lara A. B. C. ; Barreto, Matheus Q. ; Ward, Richard J.

• Document type: Journal article
• Source: BIOFUELS BIOPRODUCTS & BIOREFINING-BIOFPR; v. 16, n. 6, p. 14-pg., 2022-08-02.

Abstract

Xyloglucan (XG) is an abundant polysaccharide in plant cell walls and some seeds and requires at least four different enzymes for complete depolymerization. Recombinant xyloglucanase from Aspergillus niveus (XegA), alpha-xylosidase from Escherichia coli (Yicl), beta-galactosidase from Hypocrea jecorina (Bga1) and beta-glucosidase from H. jecorina (Bgl1) were covalently immobilized individually and in combination on chitosan-coated ferromagnetic iron oxide nanoparticles functionalized with glutaraldehyde. All immobilized enzymes presented reduced specific catalytic activity, where immobilized Yicl, XegA, Bga1 and Bgl1 retained 14.9, 76.9, 29.5 and 6.6% activity as compared with the free enzymes, respectively. Immobilized and free enzymes presented similar optimum catalytic pH and optimum catalytic temperatures differed by +/- 10 degrees C between the free and immobilized enzymes. Bga1 and Bgl1 presented decreased maximum catalytic temperatures (T-opt), while Yicl presented an increased T-opt. The T(opt )of XegA remained unaltered. Mass spectrometry confirmed that nanoparticles carrying all four co-immobilized enzymes degraded XG to glucose, galactose and xylose, and higher proportions of co-immobilized Bg11 and XegA resulted in higher XG saccharification. Although levels of Bg11 activity were limiting, five re-use cycles of the co-immobilized enzymes were demonstrated, providing proof-of-principle for the use of a four-component multienzyme nanoparticle in the breakdown of a complex polysaccharide. (C) 2022 Society of Chemical Industry and John Wiley & Sons, Ltd. (AU)

• FAPESP's process: 16/24139-6 - Design and Characterization of Multifunctional Xylanolytic Enzymes
• Grantee: Richard John Ward
• Support Opportunities: Regular Research Grants
• FAPESP's process: 17/14452-1 - A nanostructured biomimetic system for enzymatic hydrolysis of xyloglucan
• Grantee: Jéssica de Moura Soares
• Support Opportunities: Scholarships in Brazil - Master
• 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: 12/24147-8 - Rational design of multifunctional enzymes for depolimerization of lignocelulosic material
• Grantee: Lara Aparecida Buffoni de Campos Carneiro
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 16/15708-7 - Characterization of novel hydrolysis products from the synergistic hydrolysis of xyloglucan by a beta-galactosidase/alpha-xylosidase mixture
• Grantee: Lara Aparecida Buffoni de Campos Carneiro
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate