Application of Used Tea as Solid Matrix
for Immobilization of Alkaline Protease
by OVAT Method
Tapasi Polley and Uma Ghosh
Abstract Enzyme immobilization on a solid matrix is nowadays widely employed,
particularly in the food and pharmaceutical industries, in biomedical applications
such as biosensors, immunological test systems, and in the production of fine chemicals. The conventional method of enzyme immobilization includes adsorption due
to electrostatic and hydrophobic adhesion and covalent binding by internal crosslinkage of enzymes on a solid support. The immobilization of enzymes can offer
several advantages including repeated usage of enzyme thereby reducing the cost of
the enzyme, ease of product separation, and improvement in enzyme stability. The
used tea, an abundant renewable source, can be applied as solid matrix for enzyme
immobilization. In the present investigation, the immobilization of alkaline protease
was studied on used tea. Immobilized enzyme showed better thermal stability than the
free enzyme. Optimum conditions of immobilization investigated include temperature, pH and storage stability, and also reusability. The best result of immobilization
yield (34%) and immobilization efficiency (45%) was found with 300 mg of solid
matrix. The immobilized enzyme was activity even after eight cycles of repeated use.
Keywords Alkaline protease · Immobilization · Thermostability · Solid matrix
1 Introduction
Enzyme has been immobilized on several natural and synthetic supports (Naghshbandi et al. 2018). For effective immobilization, the carrier solid matrix should have
very good inertness, regenerability, physical strength and resistance to microbial
attack, thermal stability, and huge accessible surface area (Santos et al. 2015). The
main advantage of enzyme immobilization is improved stability and resistance of
T. Polley · U. Ghosh (B)
Department of Food Technology and Bio-Chemical Engineering, Jadavpur University, Kolkata
700032, India
e-mail: ughoshftbe@yahoo.co.in
T. Polley
e-mail: tapasipolley@gmail.com
© Springer Nature Singapore Pte Ltd. 2021
D. Ramkrishna et al. (eds.), Advances in Bioprocess Engineering and Technology,
Lecture Notes in Bioengineering,
https://doi.org/10.1007/978-981-15-7409-2_22
219
for Immobilization of Alkaline Protease
by OVAT Method
Tapasi Polley and Uma Ghosh
Abstract Enzyme immobilization on a solid matrix is nowadays widely employed,
particularly in the food and pharmaceutical industries, in biomedical applications
such as biosensors, immunological test systems, and in the production of fine chemicals. The conventional method of enzyme immobilization includes adsorption due
to electrostatic and hydrophobic adhesion and covalent binding by internal crosslinkage of enzymes on a solid support. The immobilization of enzymes can offer
several advantages including repeated usage of enzyme thereby reducing the cost of
the enzyme, ease of product separation, and improvement in enzyme stability. The
used tea, an abundant renewable source, can be applied as solid matrix for enzyme
immobilization. In the present investigation, the immobilization of alkaline protease
was studied on used tea. Immobilized enzyme showed better thermal stability than the
free enzyme. Optimum conditions of immobilization investigated include temperature, pH and storage stability, and also reusability. The best result of immobilization
yield (34%) and immobilization efficiency (45%) was found with 300 mg of solid
matrix. The immobilized enzyme was activity even after eight cycles of repeated use.
Keywords Alkaline protease · Immobilization · Thermostability · Solid matrix
1 Introduction
Enzyme has been immobilized on several natural and synthetic supports (Naghshbandi et al. 2018). For effective immobilization, the carrier solid matrix should have
very good inertness, regenerability, physical strength and resistance to microbial
attack, thermal stability, and huge accessible surface area (Santos et al. 2015). The
main advantage of enzyme immobilization is improved stability and resistance of
T. Polley · U. Ghosh (B)
Department of Food Technology and Bio-Chemical Engineering, Jadavpur University, Kolkata
700032, India
e-mail: ughoshftbe@yahoo.co.in
T. Polley
e-mail: tapasipolley@gmail.com
© Springer Nature Singapore Pte Ltd. 2021
D. Ramkrishna et al. (eds.), Advances in Bioprocess Engineering and Technology,
Lecture Notes in Bioengineering,
https://doi.org/10.1007/978-981-15-7409-2_22
219