Purification and Characterisation of a Newly Isolated Stable Long-Life Tannase produced by F. subglutinans (Wollenweber and Reinking) Nelson et al.

Abstract

A stable long-life tannase was synthesised by Fusarium subglutinans and the fermentation processing parameters were optimised. Maximum enzyme production (9.38 U/ml) was recorded after 96 h of incubation at 35°C, initial pH 5, in submerged culture (200 rpm) utilising 2% (w/v) tannic acid as a sole carbon source. The tannase produced was purified to electrophoretic homogeneity through two-step column chromatography and the purified form remained stable in a pH range of 6-8. Its midpoint of thermal inactivation (T m ) was recorded at 70°C after 60 min of exposure. Maximum tannase activity was enabled at pH 6 and 40°C. Ca 2+ , K + , Mg 2+ and Mn + showed a stimulatory effect while Ba 2+ , Co 2+ , Cu 2+ , Fe 3+ and Zn + showed a competitive inhibitory effect on enzyme activity. Values of K m , V max , K cat and the molecular mass of the purified enzyme were 0.116 μM ml −1 min −1 , 3.57 mM, 1.16 μM ml −1 min −1 and 150 kDa, respectively. The participation of the SH group and carbohydrates in the enzyme structure was also suggested by the results. The stability of the purified and partially purified enzyme at −15°C extended to 13 months.

Purification and Characterization of Tannin Acyl Hydrolase Produced by Mixed Solid State Fermentation of Wheat Bran and Marigold Flower byPenicillium notatumNCIM 923

“…Rajkumar and Nandy obtained a yield of 69% with ammonium sulphate precipitation [ 18 ]. Hamdy reported that 65% ammonium sulphate was optimum for tannase precipitation [ 19 ]. Sabu et al [ 20 ] found 4-fold purification yield of the dialyzed tannase with specific activity of 0.916 U/mg protein.…”

Characterization of a glucose- and solvent-tolerant extracellular tannase from Aspergillus phoenicis

“…However, the value of V max /K m was better for methyl-gallate, revealing that the catalytic cycle is faster in this substrate than in the presence of tannic acid and propyl-gallate. The affinity of the extracellular enzyme from A. phoenicis to tannic acid was two times higher than that observed for the enzyme produced by Fusarium subglutinans [39], and 19 times higher than that obtained for the enzyme from P. variable [33]. In the presence of methyl gallate, the affinity was similar to the enzyme from P. variable [33] but lower when compared to the tannases from A. niger [24] and A. oryzae [40], among others.…”

Biotechnological Advances and Challenges of Tannase: An Overview

“…The addition of Mg +2 increased the tannase activity, but it is still unclear whether if it acts as a cofactor or if it has a nonspecific effect (Kar et al 2003;Mukherjee and Banerjee 2006;Naidu et al 2008;Hamdy 2008) …”