Abstract

PRODUCTION & PURIFICATION OF A LOW MOLECULAR WEIGHT EXTRACELLULAR TANNASE FROM ASPERGILLUS TPF-4

Sunny Dhiman*, Gunjan Mukherjee and Anu Kumar

Abstract

In the present investigation, tannase production potential of Aspergillus spp. designated as Aspergillus TPF-4 was assessed in submerged culture utilizing tannic acid as substrate. Assessment of enzyme activity revealed that Aspergillus TPF-4 strain possessed the proficiency of producing a higher titer of extracellular tannase. The potential of ammonium sulphate precipitation & anion exchange chromatography for tannase purification was evaluated. The crude enzyme exhibited maximum activity at 20% saturation concentration of ammonium sulphate. Precipitation of tannase protein was achieved at 50 % saturation concentration of ammonium sulphate. The concentrated enzyme flaunted a purification fold of 28.5 with 11.40% yield upon purification by anion-exchange chromatography using DEAE-cellulose column. Purified tannase protein unveiled a single band with an apparent molecular weight of approximately 35.0kDa upon SDS-PAGE analysis. A single protein band verifies its homogeneity and complete purification.

Keywords: Aspergillus TPF-4, Tannase, Purification, Anion exchange chromatography, Submerged fermentation, SDS-PAGE.