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Immobilization of A. oryzae β-galactosidase on Silica Nanoparticles: Development of an Effective Biosensor for Determination of Lactose in Milk Whey

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Recent advances in Applied Microbiology

Abstract

The present study demonstrates the covalent immobilization of β-galactosidase on functionalized silica nanoparticles for its application in lactose and whey hydrolysis. Under optimal conditions of 1% (w/v) glutaraldehyde, protein to carrier ratio of 66.6 mg/g and pH 7.0, a very high immobilization efficiency of 94% was obtained. The pH and temperature optimum of the immobilized β-gal was 4.5 and 50 °C with ONPG as substrate. Compared to the soluble enzyme, covalently bonded nanosilica-β-gal conjugate exhibited greater stability against inhibition by galactose and a higher thermal stability at 40 °C with a t 1/2 of 15.8 h. A lower K m and increased catalytic efficiency indicated higher substrate affinity and reactivity upon enzyme attachment to nanoparticle surface. Reusability of the immobilized preparation extended up to 14 cycles. The immobilized preparation effectively hydrolyzed whey and lactose to soluble simple sugars with 50% of hydrolysis occurring in 6 h. The rate of lactose and whey hydrolysis by immobilized β-gal was 1.5 and 2.5 times higher than that for the free enzyme, respectively. Immobilized β-gal preparation may be advantageously and commercially explored for effective bioremediation of dairy waste, devising biosensors or analytical tools for food and environmental technology or conversion of whey into value-added products.

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Acknowledgments

The financial support provided by the Department of Biotechnology (Government of India) is gratefully acknowledged. Author Dr. Rajeshwari Sinha is grateful to the Council of Scientific and Industrial Research (CSIR) for research fellowship.

Conflict of Interest

The authors declare that there is no conflict of interest.

Author Contributions

Conceived and designed the experiments: AG, RS, and SKK. Performed the experiments: AG. Analyzed the data: AG, RS, and SKK. Contributed to the writing of the manuscript: AG, RS, and SKK. All authors have approved of the article.

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Authors and Affiliations

  1. Enzyme and Microbial Biochemistry Laboratory, Department of Chemistry, Indian Institute of Technology, Hauz Khas, New Delhi, 110016, India

    Anchal Goel, Rajeshwari Sinha & Sunil K. Khare

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  1. Anchal Goel
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  2. Rajeshwari Sinha
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  3. Sunil K. Khare
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Correspondence to Sunil K. Khare .

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Editors and Affiliations

  1. Department of Microbiology, Maharshi Dayanand University, Rohtak, Haryana, India

    Pratyoosh Shukla

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Goel, A., Sinha, R., Khare, S.K. (2017). Immobilization of A. oryzae β-galactosidase on Silica Nanoparticles: Development of an Effective Biosensor for Determination of Lactose in Milk Whey. In: Shukla, P. (eds) Recent advances in Applied Microbiology . Springer, Singapore. https://doi.org/10.1007/978-981-10-5275-0_1

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