Purification and Characterization of a Thermostable Tannase from Enterobacter hormaechei Z8B-60 Isolated from Slaughterhouse Waste Soil

Authors:

Sharadamma Narayanaswamy ^1*, Nagesh Babu Rangappa², Debasree Basu Mukherjee¹  

¹Department of Biochemistry, School of Allied Health Sciences, REVA University

²Department of Biochemistry, School of Sciences, Maharani Cluster University

Abstract - The present investigation aimed to identify and characterize a novel tannic acid–degrading bacterium, Enterobacter hormaechei Z8B-60, capable of producing an industrially significant tannase enzyme. The strain was isolated from slaughterhouse waste soil and screened for tannase production using Minimal Salt Medium supplemented with tannic acid. Enzymatic degradation of tannic acid into gallic acid was confirmed through thin-layer chromatography. Purification of tannase was carried out using DEAE-cellulose ion-exchange chromatography, while enzyme homogeneity and molecular characterization were analyzed by SDS-PAGE. The purified tannase exhibited a specific activity of 24.86 U/mg with optimum activity at pH 6.5 and 50 °C, indicating thermostable properties suitable for industrial applications. Kinetic analysis using methyl gallate and propyl gallate as substrates revealed Km and Vmax values of 9.26 mM and 0.66 U/mL, and 6.81 mM and 0.18 U/mL, respectively. Furthermore, Mn²⁺, Fe²⁺, and Cu²⁺ ions demonstrated modulatory effects on enzyme activity, suggesting their role in catalytic regulation. This study represents the first report of tannase production by E. hormaechei Z8B-60 and highlights its potential for biotechnological applications, particularly in the bioconversion of tannin-rich waste into commercially valuable gallic acid.

Key Words:  Enterobacter hormaechei Z8B-60, Tannin acyl hydrolase, Purification, Characterization, Enzyme stability, Industrial application