Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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A Novel Tannase from the Xerophilic Fungus Aspergillus niger GH1

  • Marco, Mata-Gomez (Food Research Department, School of Chemistry, Universidad Autonoma de Coahuila, Blvd. Venustiano Carranza and J. Cardenas s/n, ZIP) ;
  • Rodriguez, Luis V. (School of Biochemical Engineering, Universidad Autonoma de Tamaulipas, Blvd. Enrique Cardenas Gonzalez) ;
  • Ramos, Erika L. (Food Research Department, School of Chemistry, Universidad Autonoma de Coahuila, Blvd. Venustiano Carranza and J. Cardenas s/n, ZIP) ;
  • Renovato, Jacqueline (Food Research Department, School of Chemistry, Universidad Autonoma de Coahuila, Blvd. Venustiano Carranza and J. Cardenas s/n, ZIP) ;
  • Cruz-Hernandez, Mario A. (Food Research Department, School of Chemistry, Universidad Autonoma de Coahuila, Blvd. Venustiano Carranza and J. Cardenas s/n, ZIP) ;
  • Rodriguez, Raul (Food Research Department, School of Chemistry, Universidad Autonoma de Coahuila, Blvd. Venustiano Carranza and J. Cardenas s/n, ZIP) ;
  • Contreras, Juan (Food Research Department, School of Chemistry, Universidad Autonoma de Coahuila, Blvd. Venustiano Carranza and J. Cardenas s/n, ZIP) ;
  • Aguilar, Cristobal N. (Food Research Department, School of Chemistry, Universidad Autonoma de Coahuila, Blvd. Venustiano Carranza and J. Cardenas s/n, ZIP)
  • Published : 2009.09.30
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Abstract

Aspergillus niger GH1 previously isolated and identified by our group as a wild tannase producer was grown under solid-state (SSC) and submerged culture (SmC) conditions to select the enzyme production system. For tannase purification, extracellular tannase was produced under SSC using polyurethane foam as the inert support. Tannase was purified to apparent homogeneity by ultrafiltration, anion-exchange chromatography, and gel filtration that led to a purified enzyme with a specific activity of 238.14 IU/mg protein with a final yield of 0.3% and a purification fold of 46. Three bands were found on the SDS-PAG with molecular masses of 50, 75, and 100 kDa. PI of 3.5 and 7.1% N-glycosylation were noted. Temperature and pH optima were 600e and 6.0 [methyl 3,4,5-trihydroxybenzoate (MTB) as substrate], respectively. Tannase was found with a $K_M$ value of $0.41{\times}10^{-4}M$ and the value of $V_{max}$ was $11.03{\mu}$moL/min at $60^{\circ}C$ for MTB. Effects of several metal salts, solvents, surfactants, and typical enzyme inhibitors on tannase activity were evaluated to establish the novelty of the enzyme. Finally, the tannase from A. niger GH1 was significantly inhibited by PMSF (phenylmethylsulfonyl fluoride), and therefore, it is possible to consider the presence of a serine or cysteine residue in the catalytic site.

Keywords

References

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