TY - JOUR
T1 - Purification and characterization of tannase from Paecilomyces variotii
T2 - Hydrolysis of tannic acid using immobilized tannase
AU - Mahendran, B.
AU - Raman, N.
AU - Kim, D. J.
N1 - Funding Information:
Acknowledgements The senior research fellowship from the Council of Scientific and Industrial Research (CSIR), New Delhi, India, to first author is gratefully acknowledged.
PY - 2006/4
Y1 - 2006/4
N2 - An extracellular tannase (tannin acyl hydrolase) was isolated from Paecilomyces variotii and purified from cell-free culture filtrate using ammonium sulfate precipitation followed by ion exchange and gel filtration chromatography. Fractional precipitation of the culture filtrate with ammonium sulfate yielded 78.7% with 13.6-folds purification, and diethylaminoethyl- cellulose column chromatography and gel filtration showed 19.4-folds and 30.5-folds purifications, respectively. Molecular mass of tannase was found 149.8 kDa through native polyacrylamide gel electrophoresis (PAGE) analysis. Sodium dodecyl sulphate-PAGE revealed that the purified tannase was a monomeric enzyme with a molecular mass of 45 kDa. Temperature of 30 to 50°C and pH of 5.0 to 7.0 were optimum for tannase activity and stability. Tannase immobilized on alginate beads could hydrolyze tannic acid even after extensive reuse and retained about 85% of the initial activity. Thin layer chromatography, high performance liquid chromatography, and 1H-nuclear magnetic resonance spectral analysis confirmed that gallic acid was formed as a byproduct during hydrolysis of tannic acid.
AB - An extracellular tannase (tannin acyl hydrolase) was isolated from Paecilomyces variotii and purified from cell-free culture filtrate using ammonium sulfate precipitation followed by ion exchange and gel filtration chromatography. Fractional precipitation of the culture filtrate with ammonium sulfate yielded 78.7% with 13.6-folds purification, and diethylaminoethyl- cellulose column chromatography and gel filtration showed 19.4-folds and 30.5-folds purifications, respectively. Molecular mass of tannase was found 149.8 kDa through native polyacrylamide gel electrophoresis (PAGE) analysis. Sodium dodecyl sulphate-PAGE revealed that the purified tannase was a monomeric enzyme with a molecular mass of 45 kDa. Temperature of 30 to 50°C and pH of 5.0 to 7.0 were optimum for tannase activity and stability. Tannase immobilized on alginate beads could hydrolyze tannic acid even after extensive reuse and retained about 85% of the initial activity. Thin layer chromatography, high performance liquid chromatography, and 1H-nuclear magnetic resonance spectral analysis confirmed that gallic acid was formed as a byproduct during hydrolysis of tannic acid.
UR - http://www.scopus.com/inward/record.url?scp=33645229781&partnerID=8YFLogxK
U2 - 10.1007/s00253-005-0082-y
DO - 10.1007/s00253-005-0082-y
M3 - Article
C2 - 16133325
AN - SCOPUS:33645229781
SN - 0175-7598
VL - 70
SP - 444
EP - 450
JO - Applied Microbiology and Biotechnology
JF - Applied Microbiology and Biotechnology
IS - 4
ER -