• KSBB Journal
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• 제11권5호
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• pp.565-571
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• 1996

Aspergillus oryzae A Tee 2114를 사용하여 $\beta$-mannanase의 생산에 영향을 주는 배지성분의 최적 화를 수행하였다. 탄소원인 locust bean gum의 농 도를 달리하여 발효를 수행하였다. Locust bean gum의 농도가 20 g/L 이상일 때는 초기의 점도가 높아 혼합이 거의 되지 않아 초기에 $\beta$-mannanase 의 생산이 지 연되는 현상이 나타났고. Locust bean g gum의 농도가 증가할수록 $\beta$-mannanase의 역가와 균체농도가 비례척으로 증가하였다. Locust bean gum 10 g/L 배지에서 여러 가지 질소원이 $\beta$-mann anase의 생산에 미치는 영향을 살펴 본 결과,B-man n nanase의 역가는 무기질소원 중에서는 $(NH-4)_2SO_4$가 좋았으며 유기질소원 중에서는 malt extract가 가장 좋았다. 여러 가지 무기엽의 최적화를 수행한 결과 KH,PO.가 $\beta$-mannanase의 생산에 중요한 인자임을 알 수 있었다. 배지최적화 결과 최적배지 로 locust bean gum 10 giLt malt extract 3 g/L, $(NH-4)_2SO_4 2 g/L, KH_2PO_4$ 10 g/L이 결정되었으며 이때 $\beta$mannanase의 역가는 거의 6 unit/mL에 접 근하였다. 최적배지를 사용하여 발효조에서 배양을 수행하였다. 발효조의 흔합효과로 배양초기에 나타 나는 $\beta$-mannanase 생산 지 연현상을 감소시 킬 수 있었고 배양시간도 단축할 수 있었다. 27시간 배양 한 후 $\beta$mannanase의 역가 9.7 unit/mL와 비 $\beta$-mannanase의 역가 1.9 unit/mg-cell-을- 얻었다. 이 때, 생산성은 0.36 unit/mL. h이었다.

• 한국미생물·생명공학회지
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• 제25권2호
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• pp.212-217
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• 1997

A strain of Bacillus sp. WS-14 was isolated from soil. Medium optimization for ${\beta}-mannanase$ production by Bacillus sp. WS-14 was performed. Effect of various carbon sources on ${\beta}-mannanase$ production was investigated and locust bean gum was the most effective for ${\beta}-mannanase$ production. ${\beta}-mannanase$ activity and cell growth increased with increasing the concentration of locust bean gum, however, the amounts were not significant. Among nitrogen sources, soytone was the most effective for ${\beta}-mannanase$ production. Inorganic compounds such as $KH_2PO_4,\;NaCl\;Na_2CO_3\;and\;MgSO_4{\cdot}7H_2O\;on\;{\beta}-mannanase$ production were optimized for ${\beta}-mannanase$ production. Locust bean gum of 10.0 g/l, soytone of 5.0 g/l, $KH_2PO_4$ of 2.0 g/l, NaCl of 10.0 g/l, $MgSO_4{\cdot}7H_2O\;of\;0.2\;g/l,\;Na_2CO_3$, of 2.0 g/l were selected as optimum content. Production of ${\beta}-mannanase$ by using the optimum medium was carried out. The maximum ${\beta}-mannanase$ activity of 20.8 unit/ml could be obtained after 14 h fermentation which corresponed to the productivity of ${\beta}-mannanase$ of 1.48 unit/ml-h.

• 한국미생물·생명공학회지
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• 제31권1호
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• pp.57-62
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• 2003

토양으로부터 $\beta$-mannanase활성이 우수한 균주를 분리하여 형태학적, 생화학적 동정과정을 거쳐 Bacillus subtilis JS-1으로 동정하였다. 분리균이 생산하는 $\beta$-mannanase 효소의 최적활성은 55$^{\circ}C$와 pH 5.0이었다. 탄소원이 다른 배지에서 배양한 분리 균주의 상등액을 전기영동하여 효소활성을 관찰한 결과 탄소원에 상관없이 분자량 130kDa에 해당하는 단일 단백질만이 효소 활성을 나타내었다 Bacillus subtilis JS-1은 탄소원으로 lactose와 locust bean gum이 존재할 때 $\beta$-mannanase 생산성이 크게 증가하는 것으로 나타났으며, lactose와 locust bean gum이 각각 0.5 % 존재할 때 배양 상등액의 $\beta$-mannanase 활성은 30U/ml과 45U/ml로 탄소원이 없는 대조구에 비해 최대 18배 정도 생산성이 증가하였다. 배지에 locust bean gum을 첨가하였을 때 효소 생산성 뿐만 아니라 균체의 성장도 함께 증가하는 것으로 보아 분리균주는 locust bean gum을 분해하여 에너지원으로 이용하는 것으로 판단된다

• 한국미생물·생명공학회지
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• 제29권4호
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• pp.201-205
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• 2001

자연계로부터 locust bean gum이 들어 있는 선택배지에서 투명환을 형성하는 mannan 분해능을 갖는 미생물을 스크링 하였고 이를 Aeromonas sp. 로 동정하였다. Aero monas sp. 로부터 염색체 DNA를 분리하여 EcoRI으로 절단하여 대장균에 형질전환한 후 대장균 콜로니 주위에서 locust bean gum을 분해하여 투명환을 형성하는 대장균을 찾아냈다. 형질전환된 대장균으로부터 플라스미드를 분리하여 동일한 제한효소를 처리하여 확인한 결과 10 kd의 Aeromonas sp. 염섹체 DNA에 $\beta$-mannanase 유전자 존재하였다. 대장균에서 발현된 $\beta$-mannanase 의최적 반응 pH와 최적 반응온도는 각각 6.0과 $50^{\circ}C$로서 모균인 Aeromonas sp. 와 동일하였다.

• Food Science and Biotechnology
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• 제15권5호
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• pp.820-823
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• 2006

Five oligosaccharides were isolated from the hydrolysate of konjac (Amorphophallus konjac) glucomannan by a purified ${\beta}$-mannanase from Trichoderma sp. These oligosaccharides were identified as M-M, G-M, M-G-M, M-G-M-M, and M-G-G-M; where G- and M- represent ${\beta}$-1,4-D-glucopyranosidic and ${\beta}$-1,4-D-mannopyranosidic linkages, respectively. The mode of action of the mannanase on the glucomannan is discussed on the basis of the structure of the above oligosaccharides.

• Asian-Australasian Journal of Animal Sciences
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• 제27권5호
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• pp.700-705
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• 2014

A 12-week feeding trial was conducted with 87 g rainbow trout to evaluate the effects on growth performances, feed efficiency and nutrient digestibility of adding ${\beta}$-mannanase and ${\alpha}$-galactosidase enzymes, solely or in combination. Seven diets were prepared by adding ${\beta}$-mannanase, ${\alpha}$-galactosidase and mixed enzyme at two different levels (1 g/kg and 2 g/kg) to control diet (without enzyme) including soybean meal. Mixed enzymes (1 g/kg, 2 g/kg) were prepared by adding ${\beta}$-mannanase and ${\alpha}$-galactosidase at the same doses (0.5+0.5 g/kg and 1+1 g/kg). At the end of the experiment, addition of ${\beta}$-mannanase, ${\alpha}$-galactosidase and mixed enzyme to diet containing 44% soybean meal had no significant effects on growth performance and gain:feed (p>0.05). In addition, adding ${\beta}$-mannanase, ${\alpha}$-galactosidase and mixed enzyme in different rations to trout diets had no affect on nutrient digestibility and body composition (p>0.05).

• Journal of Microbiology and Biotechnology
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• 제22권3호
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• pp.331-338
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• 2012

A novel gene coding for an endo-${\beta}$-1,4-mannanase (manA) from Bacillus subtilis strain G1 was cloned and overexpressed in P. pastoris GS115, and the enzyme was purified and characterized. The manA gene consisted of an open reading frame of 1,092 nucleotides, encoding a 364-aa protein, with a predicted molecular mass of 41 kDa. The ${\beta}$-mannanase showed an identity of 90.2-92.9% ${\leq}95%$) with the corresponding amino acid sequences from B. subtilis strains deposited in GenBank. The purified ${\beta}$-mannanase was a monomeric protein on SDS-PAGE with a specific activity of 2,718 U/mg and identified by MALDI-TOF mass spectrometry. The recombinant ${\beta}$-mannanase had an optimum temperature of $45^{\circ}C$ and optimum pH of 6.5. The enzyme was stable at temperatures up to $50^{\circ}C$ (for 8 h) and in the pH range of 5-9. EDTA and most tested metal ions showed a slightly to an obviously inhibitory effect on enzyme activity, whereas metal ions ($Hg^{2+}$, $Pb^{2+}$, and $Co^{2+}$) substantially inhibited the recombinant ${\beta}$-mannanase. The chemical additives including detergents (Triton X-100, Tween 20, and SDS) and organic solvents (methanol, ethanol, n-butanol, and acetone) decreased the enzyme activity, and especially no enzyme activity was observed by addition of SDS at the concentrations of 0.25-1.0% (w/v) or n-butanol at the concentrations of 20-30% (v/v). These results suggested that the ${\beta}$-mannanase expressed in P. pastoris could potentially be used as an additive in the feed for monogastric animals.

• Asian-Australasian Journal of Animal Sciences
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• 제26권4호
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• pp.579-587
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• 2013

A total of 288 crossbred (Duroc${\times}$Landrace${\times}$Yorkshire) growing pigs were used in two experiments to investigate the effects of adding ${\beta}$-mannanase to corn-soybean meal-based diets on pig performance and apparent total tract digestibility (ATTD). Both experiments lasted 28 d and were split into two phases namely 1 to 14 days (phase 1) and 15 to 28 days (phase 2). In Exp. 1,144 pigs weighing $23.60{\pm}1.59$ kg BW were assigned to one of four corn-soybean meal-based diets containing 0, 200, 400 or 600 U/kg ${\beta}$-mannanase. Increasing the level of ${\beta}$-mannanase increased weight gain (quadratic effect; p<0.01) and feed efficiency (linear and quadratic effect; p<0.01) during the second phase and the overall experiment. However, performance was unaffected (p>0.05) by treatment during phase 1. Increasing the amount of ${\beta}$-mannanase in the diet improved (linear and quadratic effect; p<0.05) the ATTD of CP, NDF, ADF, calcium, and phosphorus during both phases. Based on the results of Exp. 1, the optimal supplementation level was determined to be 400 U/kg and this was the level that was applied in Exp. 2. In Exp. 2, 144 pigs weighing $23.50{\pm}1.86$ kg BW were fed diets containing 0 or 400 U/kg of ${\beta}$-mannanase and 3,250 or 3,400 kcal/kg digestible energy (DE) in a $2{\times}2$ factorial design. ${\beta}$-Mannanase supplementation increased (p<0.01) weight gain and feed efficiency while the higher energy content increased (p<0.01) feed intake and feed efficiency during both phases and overall. Increased energy content and ${\beta}$-mannanase supplementation both increased (p<0.05) the ATTD of DM, CP, NDF, ADF, phosphorus, and GE during both phases. There were no significant interactions between energy level and ${\beta}$-mannanase for any performance or digestibility parameter. In conclusion, the ${\beta}$-mannanase used in the present experiment improved the performance of growing pigs fed diets based on corn and soybean. The mechanism through which the improvements were obtained appears to be related to improvements in ATTD.

• Journal of Microbiology and Biotechnology
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• 제6권2호
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• pp.86-91
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• 1996

Thermostable $\beta$-mannanase from Bacillus sp. YA-14 was purified by acetone precipitation, CM-cellulose, Sephadex G-100 and hydroxyapatite column chromatography from culture supernatant. The final enzyme preparation appeared to be homogeneous on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). $\beta$-Mannanase appeared to be a monomeric protein with a molecular weight of 67, 000 daltons. The optimal pH and temperature of the enzyme reaction were pH 6.0 and $75^{\circ}C$ , respectively. The enzyme was stable at a pH range of 6.0 to 9.0 and at temperatures between 45 and $85^{\circ}C$. The kinetic constants of $\beta$-mannanase as determined with a galactomannan (locust bean) as substrate were a Vmax of 25 unit/ml and a Km of 1.1 mg/ml. The enzyme had only limited activity on galactomannan substrate. It was suggested that mg $\beta$-mannanase activity is limited by the number of branched $\alpha$-galactose residues.

• Asian-Australasian Journal of Animal Sciences
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• 제31권4호
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• pp.564-568
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• 2018

Objective: This experiment was conducted to determine the effects of dietary ${\beta}$-mannanase on the additivity of true metabolizable energy (TME) and nitrogen-corrected true metabolizable energy ($TME_n$) for broiler diets. Methods: A total of 144 21-day-old broilers were randomly allotted to 12 dietary treatments with 6 replicates. Five treatments consisted of 5 ingredients of corn, wheat, soybean meal, corn distillers dried grains with solubles, or corn gluten meal. One mixed diet containing 200 g/kg of those 5 ingredients also was prepared. Additional 6 treatments were prepared by mixing 0.5 g/kg dietary ${\beta}$-mannanase with those 5 ingredients and the mixed diet. Based on a precision-fed chicken assay, TME and $TME_n$ values for 5 ingredients and the mixed diet as affected by dietary ${\beta}$-mannanase were determined. Results: Results indicated that when ${\beta}$-mannanase was not added to the diet, measured TME and $TME_n$ values for the diet did not differ from the predicted values for the diet, which validated the additivity. However, for the diet containing ${\beta}$-mannanase, measured $TME_n$ value was greater (p<0.05) than predicted $TME_n$ value, indicating that the additivity was not validated. Conclusion: In conclusion, the additivity of energy values for the mixed diet may not be guaranteed if the diet contains ${\beta}$-mannanase.