• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.11
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• pp.1501-1506
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• 2011

In this study, we optimized the reaction conditions for the bioconversion of green tea using tannase, and to evaluate its radical scavenging activities. Tea catechins such as (-)-epigallocatechin gallate (EGCG) or (-)-epicatechin gallate (ECG) were hydrolyzed by tannase to produce (-)-epigallocatechin (EGC) or (-)-epicatechin (EC), respectively, and a common product, gallic acid. The bioconversion of tea catechins by tannase was increased as enzyme concentration, substrate concentration and incubation time for enzyme dose. The results indicated the optimum reaction conditions for tannase were tannase 30 U/mL (enzyme concentration) on 1% green tea (substrate concentration) for 1 hr (incubation time for enzyme). Tannase enhanced the radical-scavenging properties of green tea; the 2,2-azinobis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals scavenging abilities were significantly (p<0.001) greater for the tannase-treated green tea extract compared to the untreated green tea extract. It is reported that ECG has the greatest antioxidant activity among the catechins in green tea, and the release of gallic acid is considered to be beneficial because of its significant antioxidant potency. The results of this study suggest that the tannase-treated green tea increases antioxidant activities under optimum reaction conditions.

• Microbiology and Biotechnology Letters
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• v.18 no.6
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• pp.591-598
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• 1990

Six species under the basidiomycetes were screened for extracellular tannase (tannin acyl hydrolase EC 3.1. 1.20) production in submerged culture and Lenzites betulina was found to be most effective for the production of tannase. The optimum cultural conditions for tannase production were $25^{\circ}C$, pH 6.0 and 21 days of culture period, The efficient composition of culture medium for the production of tannase was performed in synthetic medium containing tannic acid, 2g; sucrose, 5g; bacto-peptone, 2g; ,$ KH_2PO_4, \;2g,\; MgSO_4.7H_2O \;0.5g,\; CuS0_4.5H_2O$, 2 mg; thiamine HCl, 100 ug and distilled water 100 ml, The tannase produced from Lenzites bdulin*r was 223.3 unit (umole of gaUic acidiml of brothlmin). The tannase had an optimal reaction conditions ofpH 6.0 and temperature of $40^{\circ}C$. The enzyme was stable at temperature below $40^{\circ}C$ and lost its activity by 50% above $60^{\circ}C$. And the stable pH range was 5.5 to 6.0.

• Journal of Microbiology and Biotechnology
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• v.17 no.6
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• pp.1049-1053
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• 2007

Tannase production by Aureobasidium pullulans DBS66 was optimized. The organism produced maximum tannase in the presence of 1% tannic acid after 36 h. Maximum gallic acid accumulation was observed within 36 h and tannic acid in the fermented broth was completely degraded after 42 h of growth. Glucose had a stimulatory effect on tannase synthesis at 0.1% (w/v) concentration. The organism showed maximum tannase production with $(NH_4)_2HPO_4$ as nitrogen source. Shaking speed of 120 rpm and 50-ml broth volume have been found to be suitable for maximum tannase production.

• Food Science and Biotechnology
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• v.16 no.2
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• pp.243-248
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• 2007

A biochemical characterization of the tannases from Paecilomyces variotii (produced at Unicamp), Aspergillus niger (purchased from Industrial Kerry Bio-Science) and A. niger cnpat 001 (purchased from Embrapa Agroindustrial Tropical-Brazil) was carried out. P variotii is a new strain obtained from the screening of 500 fungi that were tested for their production of tannase. The biochemical properties of this new tannase from P variotii were determined and compared with those of two other tannase preparations. The tannase produced from P. variotii showed optimum activity at pH 6.5. The functional temperature range of the tannases was from $20-70^{\circ}C$, with optima at $70^{\circ}C$ for P. variotii and at $60^{\circ}C$ for the commercially obtained tannase, whereas A. niger cnpat 001 showed optimum activity at $40^{\circ}C$. The effects of 1 mM preparations of cations and anions, inhibitors, surfactants, and chelators on the tannase activity from P. variotii were also studied.

• Journal of Microbiology and Biotechnology
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• v.20 no.4
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• pp.732-736
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• 2010

Five strains of tannic acid degrading bacteria were isolated and identified by phenotypic characterization. All the five isolates showed cell-associated activity, whereas only three showed extracellular activity. Serratia ficaria DTC, showing the highest cell-associated activity (0.29 U/l), was selected for further shake-flask studies. Tannase synthesis was growth associated and reached the peak in the late stationary phase of growth. Organic nitrogen sources enhanced the tannase production. Peak tannase production of 0.56 U/l was recorded in the medium having the initial pH of 6. The pH and temperature optima of the enzyme were found to be 8.9 and $35^{\circ}C$, respectively. This is the first report of cell-associated activity in the case of bacterial tannase. Cell-associated tannase of Serratia ficaria DTC could be industrially important from the perspective of its activity at broad temperature and pH ranges, and its unusually high activity at pH 8.9.

• Journal of Microbiology and Biotechnology
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• v.29 no.11
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• pp.1749-1759
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• 2019

Aspergillus ochraceus biofilm, developed on an inert support, can produce tannase in Khanna medium containing 1.5% (w/v) tannic acid as the carbon source, at an initial pH of 5.0, for 72 h at 28℃. Addition of 0.1% (w/v) yeast extract increased enzyme production. The enzyme in the crude filtrate exhibited the highest activity at 30℃ and pH 6.0. At 50℃, the half-life (T50) was 60 min and it was 260 min at pH 6.0. In general, addition of detergents and surfactants did not affect tannase activity significantly. Tannase has potential applications in various biotechnological processes such as the production of propyl gallate and in the treatment of tannin-rich effluents. The content of tannins and total phenolic compounds in effluents from leather treatment was reduced by 56-83% and 47-64%, respectively, after 2 h of enzyme treatment. The content of tannins and total phenolic compounds in the sorghum flour treated for 120 h with tannase were reduced by 61% and 17%, respectively. Interestingly, the same A. ochraceus biofilm was able to produce tannase for three sequential fermentative process. In conclusion, fungal biofilm is an interesting alternative to produce high levels of tannase with biotechnological potential to be applied in different industrial sectors.

• The Korean Journal of Food And Nutrition
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• v.24 no.4
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• pp.720-724
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• 2011

Differences in sensory evaluation, physicochemical properties and antioxidant activities between green tea concentrations and tannase-treated green tea concentrations were measured in this study. The results showed that pH did not differ dependent on tannase treatment but antioxidant ability were slightly increased through tannase treatment without significant($p$ <0.05). However, the turbidity of the tannase-treated green tea concentration had significant difference to the non-treated concentration. The astringency, tested via sensory evaluation, decreased significantly after tannase treatment($p$ <0.05). Traces of epicatechin gallate(ECG) and epigallocatechin gallate(EGCG) also disappeared after the tannase treatment, due to the fact that the ester linkages in the catechin structure were broken by the tannase. It was concluded that tannase treatment of tea products is a very effective process for decreasing astringency and turbidity.

• Korean Journal of Food Science and Technology
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• v.15 no.4
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• pp.333-341
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• 1983

Tannase of Aspergillus sp. AN-11 isolated from contaminated acorns was purified by a procedure involving ammonium sulfate precipitation, DEAE-cellulose column chromatography and Sephadex G-200 gel filtration. Physiochemical properties of the purified tannase was investigated. Tannase was purified about 37 folds with the yield of 49% from the culture broth of Aspergillus sp. AN-11. The purified tannase was homogeneous on polyacrylamide gel disc electrophoresis and was dissociable into two identical subunits on SDS-polyacrylamide gel electrophoresis. The molecular weight of the tannase was determined to be 200,000 by gel filtration on Sephadex G-200. The purified tannase showed a typical protein ultraviolet spectrum. The enzyme had a optimum pH 5.5 and optimum temperature at 30 to $40^{\circ}C$. The enzyme was stable at a pH range from 5.0 to 6.5 and at the temperature below $30^{\circ}C$. The enzyme was inactivated remarkably by $CuCl_2$ and $ZnCl_2. The Km value of the enzyme was $7.58{\times}10^{-4}\;M$.

• Journal of Microbiology and Biotechnology
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• v.21 no.9
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• pp.960-967
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• 2011

Tannin acyl hydrolase, also known as tannase, is an enzyme with important applications in the food, feed, pharmaceutical, and chemical industries. However, despite a growing interest in the catalytic properties of tannase, its practical use is very limited owing to high production costs. Several studies have already demonstrated the advantages of solid-state fermentation (SSF) for the production of fungal tannase, yet the optimal conditions for enzyme production strongly depend on the microbial strain utilized. Therefore, the aim of this study was to improve the tannase production by a locally isolated A. niger strain in an SSF system. The SSF was carried out in packed-bed bioreactors using polyurethane foam as an inert support impregnated with defined culture media. The process parameters influencing the enzyme production were identified using a Plackett-Burman design, where the substrate concentration, initial pH, and incubation temperature were determined as the most significant. These parameters were then further optimized using a Box-Behnken design. The maximum tannase production was obtained with a high tannic acid concentration (50 g/l), relatively low incubation temperature ($30^{\circ}C$), and unique low initial pH (4.0). The statistical strategy aided in increasing the enzyme activity nearly 1.97-fold, from 4,030 to 7,955 U/l. Consequently, these findings can lead to the development of a fermentation system that is able to produce large amounts of tannase in economical, compact, and scalable reactors.

• Journal of Microbiology and Biotechnology
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• v.22 no.2
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• pp.199-206
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• 2012

Naturally immobilized tannase (tannin acyl hydrolase, E.C. 3.1.1.20) has many advantages, as it avoids the expensive and laborious operation of isolation, purification, and immobilization, plus it is highly stable in adverse pH and temperature. However, in the case of cell-associated enzymes, since the enzyme is associated with the biomass, separation of the pure biomass is necessary. However, tannic acid, a known inducer of tannase, forms insoluble complexes with media proteins, making it difficult to separate pure biomass. Therefore, this study optimizes the production of cell-associated tannase using a "protein-tannin complex" free media. An exploratory study was first conducted in shake-flasks to select the inducer, carbon source, and nitrogen sources. As a result it was found that gallic acid induces tannase synthesis, a tryptose broth gives higher biomass, and lactose supplementation is beneficial. The medium was then optimized using response surface methodology based on the full factorial central composite design in a 3 l bioreactor. A $2^3$ factorial design augmented by 7 axial points (${\alpha}$ = 1.682) and 2 replicates at the center point was implemented in 17 experiments. A mathematical model was also developed to show the effect of each medium component and their interactions on the production of cell-associated tannase. The validity of the proposed model was verified, and the optimized medium was shown to produce maximum cell-associated tannase activity of 9.65 U/l, which is 93.8% higher than the activity in the basal medium, after 12 h at pH 5.0, $30^{\circ}C$. The optimum medium consists of 38 g/l lactose, 50 g/l tryptose, and 2.8 g/l gallic acid.