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Production and immobilization of β-galactosidase isolated from Enterobacter aerogenes KCTC2190 by entrapment method using agar-agar organic matrix

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Abstract

In the present study, Enterobacter aerogenes KCTC2190 was isolated from soil around a cattle shed area, which was capable of producing intracellular β-galactosidase. Partially purified β-galactosidase was immobilized by entrapment method in agar-agar gel matrix. Agar-agar entrapped beads were prepared by drop** the enzyme-agar solution to ice-cooled toluene-chloroform ((3:1 (v/v)). 45.88±0.11% activity of partially purified β-galactosidase was retained after immobilization (bead shape). Maximum immobilization yield was observed in the presence of 2.5% agar-agar concentration. After immobilization, optimum temperature required for the enzyme-substrate reaction was shifted from 50 to 60 °C and the optimum reaction time was shifted from 15 to 25 min. The optimum pH for both free and immobilized β-galactosidase was pH 7. Free enzyme showed lower activation energy in comparison with the immobilized one. For free as well as immobilized β-galactosidase thermal deactivation, rate constant (kd) increased with increasing temperature while the values of decimal reduction time (D-values) and half-lives (t1/2) decreased. Immobilization process increased the t1/2 and D-values of β-galactosidase while it decreased the kd. Thermostability of immobilized β-galactosidase was higher as they showed higher enthalpy (ΔΗ0) and Gibb’s free energy (ΔG0)value than those of the free β-galactosidase. The negative entropy (ΔS0) of free and immobilized β-galactosidase established that both were in a more ordered state within the temperature range (50 to 70 °C) studied. Immobilized β-galactosidase was able to retain 51.65±1.61% of its initial activity after 7 batches of enzyme-substrate reaction. Immobilized β-galactosidase showed 78.09±3.69% of its initial activity even after 40 days of storage at 4 °C.

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Acknowledgements

The authors are grateful to the School of Community Science and Technology, Indian Institute of Engineering Science and Technology, Shibpur, India, for giving facilities to pursue the research work. Authors are thankful to the Indian Council of Medical Research, New Delhi, India, for providing Senior Research Fellowship to carry out the study.

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  1. School of Community Science and Technology, Indian Institute of Engineering Science and Technology, Shibpur, Howrah, West Bengal, 711103, India

    Manisha Maity & Jayati Bhowal

  2. Department of Biotechnology, Heritage Institute of Technology, Kolkata, West Bengal, India

    Aparupa Bhattacharyya

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  1. Manisha Maity
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Experimental design, data collection, data analysis, and preparation of manuscript draft were carried out by Manisha Maity. Research conceptualization, data interpretation, and preparation of manuscript draft were performed by Aparupa Bhattacharyya. Research conceptualization and preparation of manuscript draft were carried out by Jayati Bhowal. All authors read and approved the final manuscript.

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Correspondence to Jayati Bhowal.

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Maity, M., Bhattacharyya, A. & Bhowal, J. Production and immobilization of β-galactosidase isolated from Enterobacter aerogenes KCTC2190 by entrapment method using agar-agar organic matrix. Appl Biochem Biotechnol 193, 2198–2224 (2021). https://doi.org/10.1007/s12010-021-03534-8

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