Microbial production of tannase - Aseanbiotechnology.info

similar characteristics in terms of pH andtemperature
activity andstability. When tannic acidis usedas
substrate Km values of 11.25 mM and0.048 mM were
obtainedwith Aspergillus and Penicillium, respectively.
(Yamada, Iibuchi, & Minoda, 1968; Adachi, Watanabe,
& Yamada, 1971; Libuchi, Minoda, & Yamada, 1972;
Aoki, Shinke, & Nishira, 1976; Chae & Yu, 1983).
Ramirez-Coronel, Viniegra-Gonzalez, Darvill, and
Augur (2003) produced two tannase forms by solid
state culture with molecular masses of 90 kDa and
180 kDa. The tannase hadan isoelectric point of 3 8, a
temperature optimum of 60–70 C anda pH optimum of
6 0. The substrate specificity of the tannase was
determined by HPLC analysis of tannin substrates and
products. The enzyme was able to remove gallic acid
from both condensed and hydrolysable tannins. Internal
sequences were obtainedfrom each of the gel-purified
and trypsin-digested tannase forms. The peptide sequences
obtainedfrom both forms were identical to
sequences within a b-glucosidase from Aspergillus
kawachii. The purifiedtannase was testedfor bglucosidase
activity and was shown to hydrolyse
cellobiose efficiently. However, no b-glucosidase activity
was detected when the enzyme was assayed in the
presence of tannic acid.
The effect of metal ions on tannase activity was
studied recently by Kar, Banerjee, andBhattacharyya
(2003). One mM Mg +2 or Hg + activatedtannase
activity. Ba +2 ,Ca +2 ,Zn +2 ,Hg +2 andAg + inhibited
tannase activity at 1.0 mM concentration andFe +3 and
Co +2 completely inhibitedtannase activity. Ag + ,Ba +2 ,
Zn +2 andHg +2 competitively inhibitedtannase activity.
Among the anions studied, 1 mM Br or S2O3 2
enhancedtannase activity. Tween 40 andTween 80
enhancedtannase activity whereas Tween 60 inhibited
tannase activity. Sodium lauryl sulfate and Triton X-100
inhibitedtannase activity. Urea stimulatedtannase
activity at a concentration of 1.5 M. Among the
chelators chosen for the present study, 1 mM EDTA
or 1,10-o-phenanthrolein inhibitedtannase activity.
Dimethyl sulphoxide and b-mercaptoethanol inhibited
tannase activity at 1 mM concentration whereas soybean
extract inhibitedtannase activity at concentrations
varying from 0.05% to 1.0% (w/v). Among the nitrogen
sources selectedammonium ferrous sulfate, ammonium
sulfate, ammonium nitrate andammonium chloride
enhancedtannase activity at 0.1% (w/v) concentration.
8. Tannase applications
One of the major applications of tannase is in the
manufacturing of instantaneous tea. Tannase applications
in food and beverage industrial products contribute
to remove the undesirable effects of tannins
(Boadi & Neufeld, 2001). Other important application
ARTICLE IN PRESS
of tannase is the production of gallic acid and
propylgallate (Kar, Banerjee, & Bhattacharyya, 2002).
The former one is usedin the pharmaceutical industry
for the synthesis of antibacterial drugs and in the food
industry as substrate for the chemical synthesis of food
preservatives such as pyrogallol andgallates. On the
other hand, propylgallate is a very important food
antioxidant (Sharma & Gupta, 2003). Tannase may
contribute to plant cell wall degradation by cleaving
some of the cross-links existing between cell wall
polymers (Garcia-Conesa, Ostergaard, Kauppinen, &
Williamson, 2001). Tannase can be potentially usedfor
the degradation of tannins present in the effluents of
tanneries, which represent serious environmental problems
(Van de Lagemaat & Pyle, 2001). Also, it can be
usedin the preparation of animal feeding using as
culture support the mycelial wastes from penicillin
manufacture (Nuero & Reyes, 2002).
9. Concluding remarks
The use of tannase from different microbial sources
may have benefits for different areas such as food,
beverage, cosmetic andpharmaceutical industries as
well as environmental depollution. For that, more effort
is needed in order to develop more productive process
for tannase production. In this sense, solid state
fermentation presents more advantages that the submergedtype
of culture. Improvements on tannase
immobilization are needed for the development of
cheaper process. For that, enzymatic preparations with
improved catalytic characteristics must be developed.
Acknowledgements
C.N. Aguilar thanks CONACYT-SEP (Project:
42244) andCOAH-CONACYT (COAH-2002-
CO1.2565 and4652) for financial support. The present
work was conducted within the framework of the ECOS
program (M02A02). R. Belmares andA. Ram!ırez-
Coronel are the recipients of a M.Sc. andPhD scholarships
from CONACYT-Me´ xico, respectively.
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Induction and repression patterns of fungal tannase