Purification and characterization of tannase from Paecilomyces variotii

Hydrolysis of tannic acid using immobilized tannase

B. Mahendran, N. Raman, Dong Ju Kim

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)444-450
Number of pages7
JournalApplied Microbiology and Biotechnology
Volume70
Issue number4
DOIs
Publication statusPublished - 2006 Apr 1

Fingerprint

Paecilomyces
Tannins
Hydrolases
Purification
Hydrolysis
Gels
Acids
Molecular mass
Polyacrylates
Electrophoresis
Ammonium Sulfate
tannase
Thin layer chromatography
Column chromatography
Gel Chromatography
Alginate
High performance liquid chromatography
Sodium dodecyl sulfate
Fractional Precipitation
Chromatography

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology
  • Bioengineering
  • Microbiology (medical)

Cite this

Purification and characterization of tannase from Paecilomyces variotii : Hydrolysis of tannic acid using immobilized tannase. / Mahendran, B.; Raman, N.; Kim, Dong Ju.

In: Applied Microbiology and Biotechnology, Vol. 70, No. 4, 01.04.2006, p. 444-450.

Research output: Contribution to journalArticle

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