Exploring magnetic and imprinted cross-linked enzyme aggregates of rhamnopyranosidase in microbioreactors

• Published in: Bioresource technology 2018-02, Vol.249, p.704-712
• Main Authors: Martins, Samuel L., Albuquerque, Barbara F., Nunes, Mário A.P., Ribeiro, Maria H.L.
• Format: Article
• Language: English
• Subjects: Bioreactors; Cross-Linking Reagents; Enzyme Stability; Enzymes, Immobilized; Hydrolysis; Magnetic cross-link enzyme aggregates; Magnetics; Magnetite Nanoparticles; mCLEAs; Mini-packed bed bioreactor; Operational stability; Rhamnopyranosidase
• ISSN: 0960-8524; 1873-2976
• DOI: 10.1016/j.biortech.2017.10.078

•Development of rhamnopyranosidase crosslink enzyme aggregates functionalized with magnetite.•Magnetic rhamnopyranosidase could be separated from reaction mixture by magnetic field.•4′,5,7-trihydroxyflavanone-7-rhamnoglucoside hydrolysis with rhamnopyranosidase was optimized.•Development of a mini-packed bed reactor with magnetic rhamnopyranosidase crosslink aggregates.•Magnetized rhamnopyranosidase crosslink aggregates show enhanced stability and reusability. The goal of this work was the development of magnetic cross link enzyme aggregates (mCLEAs) of rhamnopyranosidase (Rhmnase), prepared by chemical cross-linking with functionalized magnetite nanoparticles for glycompounds biosynthesis in microbioreactors (specially design 24-well microplate and mini-packed bed). Rhamnopyranosidase (EC number 3.2.1.40) present high potential in glycocompounds production, with applications in food and pharmaceutical industries. The influence of precipitants, cross-linkers, temperature and time on (m)CLEAs@Rhmnase development were optimized. Biocatalyst activity was accessed in the hydrolysis of 4′,5,7-trihydroxyflavanone-7-rhamnoglucoside and kinetic constants in the deglycosylation reaction were evaluated. Rhmnase operational stability was enhanced in mCLEAs, retaining almost 90% initial activity after 7 cycles of 24 h each. In a mini-packed bed bioreactor a maximum volumetric productivity of 140 μmol/L.h was attained. In this bioreactor the operational stability of mCLEAs@Rhmnase were evaluated at a flow rate of 5 mL/h during 5 days and a residual activity of 95% was observed.