β-Glucosidase from Streptomyces griseus: Nanoparticle immobilisation and application to alkyl glucoside synthesis

- A new β-glucosidase was cloned, expressed and purified from S. griseus.
- S. griseus β-glucosidase was stable to both elevated temperatures and solvents.
- The enzyme can be readily immobilised on Zinc nanoparticles by simple absorption.
- This glucosidase was shown to catalyse the synthesis of alkyl glucosides.

A novel β-glucosidase from Streptomyces griseus was cloned and overexpressed in E. coli. The purified β-glucosidase (44 kDa) had a Km of 8.6 ± 0.5 mM and a Vmax of 217 ± 5.0 μmoles−1min−1mg at 37 °C, pH 7.2 with p-nitrophenyl-β-D glucopyranoside as substrate. The enzyme was characterised in terms of pH optimum (pH 6.9), temperature optimum (69 °C) and the influence of solvents and effectors. Purified S. griseus β-glucosidase was successfully immobilised, by simple absorption, onto zinc oxide (ZnO) nanoparticles without covalent modification. It remained tightly bound even after extensive washing and could be reused up to ten times without significant loss of activity. The immobilised enzyme had a higher optimum temperature and greater thermostability than the free enzyme. In immobilised form the enzyme readily catalysed the synthesis of alkyl glucosides.