• 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.

• 한국식품영양과학회지
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• 제23권3호
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• pp.509-514
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• 1994

Penicillium purpurogenum , which produces a copra galactomannan degrading enzyme extracellularyl, was isolated from soil , and its properties and formation condition of mannooligosaccharides were investigated. The optimum ph and temperature for the activity of the mannanase were 5.5 and 55$^{\circ}C$, respectively. The mannanase was stable in between pH 3.5 and 7.0 after 2 hr incubation at 3$0^{\circ}C$ lost 90% of the original activity after incubation at 55$\AA$ and pH 5.5 for 2 hr. With two different substrate concentration, hydrolysis of white coprameal proceeded rapidly at the early stage of the reaction, but gradually solwed thereafter especially at a higher concentration of copra meal (20 %). The enzyme hydrolyzed white copra meal to monosaccharides, mannobiose and mannotriose at the final stage of the reaction.

• 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.

• Journal of Animal Science and Technology
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• 제55권2호
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• pp.115-122
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• 2013

This study was conducted to evaluate the effect of dietary ${\beta}$-mannanase supplementation and palm kernel meal (PKM) inclusion (5%) on laying performance, egg quality and nutrient utilizability of laying hens with 73 weeks of age. A total of 240 Lohmann brown laying hens with average 77.5% egg production were randomly allocated with 60 hens per treatment, 4 replicates per treatment and 15 hens per replicate. Experimental design was a completely randomized design with $2{\times}2$ factorial arrangement, with the factors being (1) two levels of PKM (0 vs. 5%) and (2) with or without dietary ${\beta}$-mannanase (480 IU/kg of diet CTCzyme$^{(R)}$) supplementation. All hens were housed in cages ($35cmW{\times}35cmD{\times}40cmH$) with 2 hens per cage for six weeks feeding trial. Laying performance was recorded daily during feeding trial. Egg quality, nutrients utilizability and blood assays were done at the end of feeding trial. Egg production was improved (P<0.05) by both dietary PKM inclusion and ${\beta}$-mannanase combined supplementation. Either ${\beta}$-mannanase or PKM did not affect feed intakes and feed conversion ratio of all diets. Egg weight of hens fed diet containing 5% of PKM had heavier (P<0.05) eggs compared with hens fed without PKM. Albumen height was improved (P<0.05) by dietary mannanase supplementation. Crude fat utilization of 5% PKM diet was higher than that of no PKM diet regardless of ${\beta}$-mannanase supplementation. Both DM and total carbohydrate utilization were decreased (P<0.05) in hens fed 5% PKM diet. Serum IgG and yolk IgY contents of PKM groups were lower (P<0.05) than those of no PKM groups. This result showed that 5% PKM diet, independent of dietary ${\beta}$-mannanase supplementation, was able to improve egg production. In addition, dietary ${\beta}$-mannanase supplementation could be used for improving the albumen height of eggs.

• 미생물학회지
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• 제52권3호
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• pp.336-343
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• 2016

두 종류의 mannanases를 생산하는 Cellulosimicrobium sp. YB-43로부터 mannanase 유전자를 클로닝하고 그 염기서열을 결정하였다. Mannanase 유전자는 manB로 명명되었으며, 427 아미노 잔기로 구성된 단백질을 코드하는 1,284개 염기로 구성되었다. ManB는 추론된 아미노산 배열에 근거해서 glycosyl hydrolase family 5에 속하는 mannanase와 상동성이 높은 활성영역과 함께 2개의 탄수화물 결합영역을 포함하고 있는 다영역 효소로 확인되었다. Cellulosimicrobium sp. YB-43의 manB 유전자를 함유한 재조합 대장균의 균체 파쇄상등액으로부터 정제된 ManB의 아미노 말단 배열이 QGASAASDG로 결정되었으며 이는 SignalP4.1 server로 그람 음성균을 기준으로 예측된 signal peptide의 결과와 정확하기 일치하였다. 정제된 ManB의 최적 반응조건은 $55^{\circ}C$와 pH 6.5-7.0이며 locust bean gum (LBG), konjac과 guar gum을 가수분해 하였으며, 셀룰로스, 자일란, 전분과 para-nitrophenyl-${\beta}$-mannopyranoside에 대해서는 분해활성이 없었다. ManB의 활성은 $Mg^{2+}$, $K^+$와 $Na^+$에 의해 약간 저해되었으며 $Cu^{2+}$, $Zn^{2+}$, $Mn^{2+}$과 SDS에 의해서는 크게 저해되었다. 또한 이 효소는 mannobiose 보다 큰 중합도를 갖는 만노올리고당을 가수분해하였으며, LBG와 만노올리고당을 가수분해하였을 때 mannobiose가 가장 많은 양으로 생성되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제28권7호
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• pp.1014-1019
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• 2015

The purpose of this study was to determine the effect of ${\beta}$-mannanase supplementation on digestible energy (DE) and metabolizable energy (ME) contents of copra expellers (CE) and palm kernel expellers (PKE) fed to pigs. Six barrows with an initial body weight of 38.0 kg (standard deviation = 1.5) were randomly allotted to a $6{\times}6$ Latin square design with 6 dietary treatments and 6 periods. Six experimental diets were prepared in a $3{\times}2$ factorial treatment arrangement with 3 diets of a corn-soybean meal-based diet, a CE 30% diet, and a PKE 30% diet and with 2 concentrations of supplemental ${\beta}$-mannanase at 0 or 2,400 U/kg. All diets had the same proportion of corn:soybean meal ratio at 2.88:1. The marker-to-marker procedure was used for fecal and urine collection with 4-d adaptation and 5-d collection periods. No interactive effects were observed between diet and ${\beta}$-mannanase on energy digestibility and DE and ME contents of experimental diets. However, diets containing CE or PKE had less (p<0.05) DE and ME contents compared with the corn-soybean meal-based diet. The DE and ME contents in CE and PKE were not affected by supplemental ${\beta}$-mannanase. Taken together, we failed to find the effect of ${\beta}$-mannanase supplementation on energy utilization in CE and PKE fed to pigs.

• Animal Bioscience
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• 제36권11호
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• pp.1700-1708
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• 2023

Objective: The objective of this study was to determine the effects of β-mannanase on metabolizable energy (ME) and apparent total tract digestibility (ATTD) of protein in various feedstuffs including barley, copra meal, corn, corn distillers dried grains with solubles (DDGS), palm kernel meal, sorghum, and soybean meal. Methods: A basal diet was formulated with 94.8% corn and 0.77% amino acids, minerals, and vitamins and test diets replacing corn-basal diets with barley, corn DDGS, sorghum, soybean meal, or wheat (50%, respectively) and copra meal or palm kernel meal (30%, respectively). The basal diet and test diets were evaluated by using triplicated or quadruplicated 2×2 Latin square designs consisting of 2 diets and 2 periods with a total of 54 barrows at 20.6±0.6 kg (9 wk of age). Dietary treatments were levels of β-mannanase supplementation (0 or 800 U/kg of feed). Fecal and urine samples were collected for 4 d following a 4-d adaptation period. The ME and ATTD of crude protein (CP) in feedstuffs were calculated by a difference procedure. Data were analyzed using Proc general linear model of SAS. Results: Supplementation of β-mannanase improved (p<0.05) ME of barley (10.4%), palm kernel meal (12.4%), sorghum (6.0%), and soybean meal (2.9%) fed to growing pigs. Supplementation of β-mannanase increased (p<0.05) ATTD of CP in palm kernel meal (8.8%) and tended to increase (p = 0.061) ATTD of CP in copra meal (18.0%) fed to growing pigs. Conclusion: This study indicates that various factors such as the structure and the amount of β-mannans, water binding capacity, and the level of resistant starch vary among feedstuffs and the efficacy of supplemental β-mannanase may be influenced by these factors.

• 한국동물학회:학술대회논문집
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• 한국동물학회 2000년도 한국생물과학협회 학술발표대회
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• pp.284.4-285
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• 2000

• Journal of Animal Science and Technology
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• 제59권7호
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• pp.19.1-19.10
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• 2017

Background: To reduce use of main feed ingredient like corn, soy bean meal (SBM) and wheat, alternative ingredients has been studied like copra meal (CM). Production amount of CM which has been high makes CM to be an alternative feed stuff. However, low digestibility on AA and low energy content by high fiber content can be an obstacle for using CM. This experiment was conducted to evaluate the effects of CM supplementation with ${\beta}$-mannanase on growth performance, blood profile, nutrient digestibility, pork quality and economic analysis in growing-finishing pigs. Methods: A total of 100 growing pigs ([Yorkshire ${\times}$ Landrace] ${\times}$ Duroc) averaging $31.22{\pm}2.04kg$ body weight were allotted to 5 different treatments by weight and sex in a randomized complete block (RCB) design in 5 replicate with 4 pigs per pen. Treatments were 1) Control (corn-SBM based diet + 0.1% of ${\beta}$-mannanase (800 IU)), 2) CM10 (10% copra meal + 0.1% ${\beta}$-mannanase (800 IU)), 3) CM15 (15% copra meal + 0.1% ${\beta}$-mannanase (800 IU)), 4) CM20 (20% copra meal + 0.1% ${\beta}$-mannanase (800 IU)) and 5) CM25 (25% copra meal + 0.1% ${\beta}$-mannanase (800 IU)). Four phase feeding program was used: growing I (week 1-3), growing II (week 4-6), finishing I (week 7-9) and finishing II (week 10-12). Results: In growth performance, there was no significant difference among treatments during whole experimental period. In growingI phase, G:F ratio tended to increase when CM was increased (P = 0.05), but ADG and ADFI tended to decrease in finishingII phase (linear, P = 0.08). Also, increasing CM reduced ADG (linear, P = 0.02) and feed efficiency (linear, P = 0.08) during the whole finishing period. In blood profiles, BUN was linearly increased as CM increased (linear, P = 0.02) at growingII period. In digestibility trial, there was no significant difference in dry matter, crude fat, crude ash and nitrogen digestibility. However, crude protein digestibility was decreased linearly (linear, P = 0.02). In economic analysis, feed cost per weight gain and total feed cost per pig were reduced in overall period when CM was provided by 25% (linear, P = 0.02). Conclusion: CM with 0.1% of ${\beta}$-mannanase (800 IU) could be supplemented instead of corn and SBM up to 25% without detrimental effects on growth performance and pork quality of growing-finishing pigs.

• Journal of Microbiology and Biotechnology
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• 제14권6호
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• pp.1295-1302
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• 2004

A gene encoding the mannanase of Bacillus subtilis WL-7, which had been isolated from Korean soybean paste, was cloned into Escherichia coli, and the gene product was purified from the culture filtrate of the recombinant E. coli. This mannanase gene, designated manA, consisted of 1,086 nucleotides, encoding a polypeptide of 362 amino acid residues. The deduced amino acid sequence was highly homologous to those of mannanases belonging to the glycosyl hydrolase family 26. The molecular mass of the purified mannanase was 38 kDa as estimated by SDS-PAGE. The enzyme had a pH optimum at 6.0 and a temperature optimum at $55^{\circ}C$. The enzyme was active on locust bean gum, konjak, guar gum, and lichenan, while it did not exhibit activity towards yeast mannan, laminarin, carboxymethylcellulose, $\beta$­glucan, xylan, and para-nitrophenyl-$\beta$-mannopyranoside.