Horseradish peroxidase immobilized in calcium alginate-gelatin hybrid beads with high stability against metallic ions and organic solvents

Abstract

Horseradish peroxidase (HRP) was immobilized in hybrid calcium alginate-gelatin hydrogel beads by entrapment. Then, organic solubility, metal ion compatibility, pH/temperature studies of the synthesized materials have been carried out. Hybrid beads (Ca-Alg@gel-HRP) were synthesized under optimum synthesis conditions with high yield and specific activity. The synthesized bead size was determined to be approximately 0.29 mm in diameter, hydrogel structure and transparent appearance. Accordingly, the optimum pH of free HRP and Ca-Alg@gel HRP was found to be pH 6 and 5 and the optimum temperature was found to be 40 °C and 60 °C respectively. For all identified metal ions except Fe²⁺, the Ca-Alg@gel HRP had a higher activity than the free HRP. In the presence of 1 mM Na⁺, Ca²⁺, Mg²⁺, Ba²⁺, Cu²⁺, Ni²⁺, Mn²⁺, Hg²⁺ ions, the activity increased by more than 20%. The relative activity was also approximately 153%, 150% and 145% in 2.5 mM chloroform, glycerol and DMF. Free enzyme activity was reduced by more than 30%. The Ca-Alg@gel HRP at room temperature and 4 °C retained most of its activity after 14 days. In cycle 5, it had half the activity. As a conclusion, it may have significant potential in a number of industrial processes, particularly in organic synthesis, because it is tolerant to metal ions and organic solvents and is a cheap, practical and simple immobilization method.

Keywords:

• Enzyme
• ımmobilization
• hybrid beads
• stability
• organic synthesis
• entrapment method

Disclosure statement

All authors declare that there is no potential conflict of interest.

Author contributions

K.Y.D., M.O. and C.G.S. performed experiments. K.Y.D and C.G.S. processed the experimental data and completed the analysis. O.A., K.Y.D and C.G.S. wrote the manuscript and supervised the project.

Additional information

Funding

This study supported by Scientific Research Projects Coordination Unit of Kafkas University (grant number: 2021-FM-86), Turkey.