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Immobilization of β-galactosidase from Lactobacillus plantarum HF571129 on ZnO nanoparticles: characterization and lactose hydrolysis

Abstract

β-Galactosidase from Lactobacillus plantarum HF571129 was immobilized on zinc oxide nanoparticles (ZnO NPs) using adsorption and cross-linking technique. Immobilized β-galactosidase showed broad-spectrum pH optima at pH 5–7.5 and temperature 50–60 °C. Fourier transform infrared spectroscopy (FTIR) and field emission scanning electron microscopy (FESEM) showed that β-galactosidase successfully immobilized onto supports. Due to the limited diffusion of high molecular weight substrate, K m of immobilized enzyme slightly increased from 6.64 to 10.22 mM, while V max increased from 147.5 to 192.4 µmol min−1 mg−1 as compared to the soluble enzyme. The cross-linked adsorbed enzyme retained 90 % activity after 1-month storage, while the native enzyme showed only 74 % activity under similar incubation conditions. The cross-linked β-galactosidase showed activity until the seventh cycle and maintained 88.02 % activity even after the third cycle. The activation energy of thermal deactivation from immobilized biocatalyst was 24.33 kcal/mol with a half-life of 130.78 min at 35 °C. The rate of lactose hydrolysis for batch and packed bed was found to be 0.023 and 0.04 min−1.

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Abbreviations

FTIR:

Fourier transform infrared spectroscopy

ZnO NPs:

Zinc oxide nanoparticles

GRAS:

Generally recognized as safe

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

FESEM:

Field-emission scanning electron microscopy

ONPG:

O-Nitrophenyl-β-d-galactopyranoside

K d :

Deactivation rate constants

V max :

Maximum reaction velocity

K m :

Michaelis–Menten constants

ΔH :

Enthalpy

ΔG :

Gibbs free energy

ΔS :

Entropy

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Acknowledgments

We are greatly indebted to the VIT University for the constant encouragement, help and support for extending the necessary facilities.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Correspondence to V. Mohanasrinivasan.

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Selvarajan, E., Mohanasrinivasan, V., Subathra Devi, C. et al. Immobilization of β-galactosidase from Lactobacillus plantarum HF571129 on ZnO nanoparticles: characterization and lactose hydrolysis. Bioprocess Biosyst Eng 38, 1655–1669 (2015). https://doi.org/10.1007/s00449-015-1407-6

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Keywords

  • β-Galactosidase
  • Zinc oxide nanoparticles
  • Lactose hydrolysis
  • Glutaraldehyde
  • FTIR