Particulate and spherical cellulosic supports derived from sugarcane bagasse for oriented immobilization of recombinant CBD-tagged enzymes.

Leonhardt F.; Kuhn D.; Schwingel Henn G.; de Andrade B.; Gennari A.; de Moraes Pedroso A.; Führ A.; Gregory A.; Renard G.; Chies J. et al. · Bioprocess and biosystems engineering · 2026

**Background & Methods** This study developed cellulosic supports from sugarcane bagasse using alkaline hydrolysis (L) and sequential acid-alkaline hydrolysis (LH) treatments for oriented immobilization of recombinant β-galactosidase-CBD (β-Gal-CBD) enzymes. Both particulate and spherical alginate-based supports were characterized using FTIR, DSC, and SEM, with immobilization performance and enzymatic activity evaluated across varying loading conditions. **Key Findings** • LH treatment produced the highest cellulose purity support, with the particulate LH-Gal achieving recovered activity exceeding 100% at 10 U/g loading, demonstrating enzymatic hyperactivation—a significant result for agro-industrial residue-derived supports. • Among spherical supports, LH-Sph exhibited superior yield and recovered activity at optimal conditions of 3 hours and 1 U/sphere, substantially outperforming the alginate control support, confirming that cellulosic bagasse presence is determinant for oriented immobilization. • Economic analysis demonstrated industrial viability for low-lactose milk production using LH-Gal biocatalyst across 5-40 reuse cycles, with internal rate of return of 42.8-73.1%, return on investment of 340-634%, and discounted payback period of 1.65-2.98 years. **Dosage & Administration** Optimal enzyme loading: 10 U/g for particulate supports; 1 U/sphere for spherical supports. Immobilization time: 3 hours for spherical supports. **Safety & Adverse Effects** Not reported. **Evidence Quality** This is a laboratory-based bioprocess engineering study with controlled experimental conditions but lacks clinical relevance to human cannabis use. The research focuses on enzyme immobilization technology using agro-industrial waste for industrial applications rather than cannabinoid-related therapeutic outcomes. The study demonstrates strong technical rigor in characterization methods and economic modeling; however, it contains no information on safety, efficacy, or clinical applicability to patients. The "CBD" designation refers to cellulose-binding domain, not cannabidiol, creating potential for terminology confusion. Evidence quality is high for biotechnology applications but not translatable to clinical cannabis medicine contexts.

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