• 한국식품영양과학회지
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• 제40권11호
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• pp.1501-1506
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• 2011

본 연구는 tannase를 이용하여 녹차의 생물학적 전환에 대한 최적 추출 조건을 확립하고 최적 추출 조건으로 마련된 녹차 추출물에 대해 라디칼 소거능 검증을 통해 항산화력의 향상 정도를 평가하고자 하였다. 그 결과 tannase의 반응은 0.5시간 내에 대부분 이루어지며 처리 1시간까지도 일부 작용이 이루어져 최적 추출물을 획득하기 위한 tannase의 반응 시간을 1시간으로 결정하였다. 기질 농도 1% 이상의 경우에서는 EC 및 EGC 전환율이 오히려 낮아져 tannase 작용을 위한 기질 농도는 1%가 적당한 것으로 판단되었다. 또한 tannase 농도가 증가함에 따라 EC, EGC 및 gallic acid는 증가되었으며 일정 농도(30 U/mL)에서 급격한 증가를 나타내었고, 이 농도부터는 통계적으로 EC와 EGC 전환율이 증가하지 않는 것으로 보아 tannase의 적정 반응 농도는 30 U/mL으로 판단되었다. 위의 조건으로 마련된 최적의 tannase 처리 녹차 추출물의 라디칼 소거능은 tannase 처리 전 녹차 추출물에 비해 ABTS와 DPPH 라디칼에 있어 모두 유의하게 소거능이 증가되는 것으로 나타나 tannase 처리 최적 추출 조건에 의해 녹차의 항산화력이 향상됨을 알 수 있었다.

• 한국미생물·생명공학회지
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• 제18권6호
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• pp.591-598
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• 1990

Lenzites betulina(조개껍질버섯균) 등 6종 담자균류의 tannase (tannin acylhydrolase EC 3.1.1.20) 생산을 비교하고 Lenzites betulina가 가장 우수하여 이 균주의 배양물로부터 효과적인 tannase 생산조건과 효소의 특성을 검토하였다. Lenzites betulina의 tannase 최적 생산을 위한 배양 조건은$25^{\circ}C$, pH6.0에서 21일이었고, tannase acid 2g, sucrose 5g, bacto-peptone 2g,$ KH_2PO_4, \;2g,\; MgSO_4.7H_2O \;0.5g,\; CuS0_4.5H_2O$ 2mg, thinamine.HCL 100Mug, 증류수 1000ml이었다.

• Journal of Microbiology and Biotechnology
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• 제17권6호
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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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• 제16권2호
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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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• 제20권4호
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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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• 제29권11호
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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.

• 한국식품영양학회지
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• 제24권4호
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• pp.720-724
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• 2011

녹차 농축액을 탄닌 가수분해 효소를 이용하여 가수 분해한 후 기존 녹차 농축액과 비교하여 이화학 및 관능적 검사를 실시하였다. pH, 항산화 능력은 효소분해 전후 유의적인 차이를 나타내지 않았으나, 효소 분해 후 탁도는 유의적으로 더욱 맑아졌다. 녹차 탄닌의 주성분인 카테킨류는 탄닌분해 효소에 의해 분해되어 전체적으로 감소하는 경향을 나타내었다. 갈레이트형 카테킨 성분인 에피갈로카테킨 갈레이트(EGCG)와 에피카테킨 갈레이트(ECG)는 EGC와 EC 그리고 gallic acid로 효소 분해되어 효소처리 녹차 농축액에서는 EGCG와 ECG가 측정되지 않았다. 떫은맛의 관능검사는 효소분해 후 유의적으로 떫은맛의 감소가 발생하였다. 따라서 탄닌분해 효소를 이용하여 녹차 추출물을 분해하면 녹차액은 맑아지지만, 기존 녹차 추출물의 문제점인 떫은맛은 감소되어 다양한 식품에 응용할 수 있는 것으로 나타났다.

• 한국식품과학회지
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• 제15권4호
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• pp.333-341
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• 1983

도토리 부패물로부터 분리한 Aspergillus sp, AN-11 균주가 분비하는 tannase를 일련의 ammonium S-ulfate에 의한 침전, DEAE-Cellulose Column Chromatography 및 Sephadex G-200 gel filtration 방법에 의하여 정제하여 물리화학적 성질을 규명하였다. 정제된 tannase 는 비활성(比活性)이 조(粗)tannase에 비하여 37 배정도 증가되었으며, 효소활성이 회수(回收)는 약 49%이었다. 정제된 tannase 는 polyacrylamide gel 전기영동에 의한 균일성을 나타내었고, SDS-polyacrylamide gel 전기영동에서는 단일 밴드를 형성하는 두개의 동일한 subunits로 분리되었으며, 본 도토리 tannin 분해효소의 분자량은 200,000 정도로 계산되었다. 정제된 tannase는 전형적인 단백질 UV흡수 스펙트럼을 나타내었으며, 한편 최적 pH 몇 온도는 5.5 와 $30{\sim}40^{\circ}C$이었고 pH $5.0{\sim}6.5$와 온도 $30^{\circ}C$ 이하의 범위 내에서 비교적 안정성을 보였으며, $CuCl_2$ 및 $ZnCl_2금속염에 의해서는 효소활성이 크게 저해되었다. 또한 정제된 tannase의 Km 값은 $7.58{\times}10^{-4}M$이었다.

• Journal of Microbiology and Biotechnology
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• 제21권9호
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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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• 제22권2호
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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.