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KMID : 1094720100150050793
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Biotechnology and Bioprocess Engineering
2010 Volume.15 No. 5 p.793 ~ p.799
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Purification and characterization of extracellular tannin acyl hydrolase from Aspergillus heteromorphus MTCC 8818
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Chhokar Vinod
Seema
Beniwal Vikas
Salar Raj Kumar
Nehra K. S.
Kumar Anil
Rana J. S.
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Abstract
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A tannase (E.C. 3.1.1.20) producing fungal strain was isolated from soil and identified as Aspergillus heteromorphus MTCC 8818. Maximum tannase production was achieved on Czapek Dox minimal medium containing 1% tannic acid at a pH of 4.5 and 30°C after 48 h incubation. The crude enzyme was purified by ammonium sulfate precipitation and ion exchange chromatography. Diethylaminoethyl-cellulose column chromatography led to an overall purification of 39.74-fold with a yield of 19.29%. Optimum temperature and pH for tannase activity were 50°C and 5.5 respectively. Metal ions such as Ca2+, Fe2+, Cu1+, and Cu2+ increased tannase activity, whereas Hg2+, Na1+, K1+, Zn2+, Ag1+, Mg2+, and Cd2+ acted as enzyme inhibitors. Various organic solvents such as isopropanol, isoamyl alcohol, benzene, methanol, ethanol, toluene, and glycerol also inhibited enzyme activity. Among the surfactants and chelators studied, Tween 20, Tween 80, Triton X-100, EDTA, and 1, 10-o-phenanthrolein inhibited tannase activity, whereas sodium lauryl sulfate enhanced tannase activity at 1% (w/v).
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KEYWORD
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Aspergillus heteromorphus, tannase, DEAE-cellulose, tannic acid, metal ion, organic solvent
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