• Microbiology and Biotechnology Letters
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• v.32 no.4
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• pp.347-351
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• 2004

Hydrolysis of manno-oligosaccharides and galactomannan was studied with the purified Bacillus subtilis WL-7 mannanase from recombinant Eschericoli. The predominant products of hydrolysis were mannose, mannobiose and mannotriose. The enzyme could hydrolyze $\beta$-1 A-linked manno-oligosaccharides larger than mannobiose, but was not active on mannobiose. When the mannanase hydrolyzed manno-oligo saccharides of degree of polymerization(DP) 4-6, it was more active on the substrate of higher DP. Based on analysis of transient reaction products by TLC, the enzyme was found to have a preference for internal $\beta$-IA-mannosidic linkages, which are the central mannosidic bond of mannotetraose and the two middle mannosidic bonds of mannopentaose. The $\beta$-l A-mannosidic bonds situated at the second and fourth positions from the nonreducing end of mannohexaose were preferenhydrolyzed by the mannanase. Locust bean gum(LBG) was enzymatically hydrolyzed with higher efficiency than guar gum, resulting that amount of reducing sugars was liberated more efficiently from LBG than guar gum with same activity of mannanase.

• Journal of Life Science
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• v.27 no.9
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• pp.1003-1010
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• 2017

An open reading frame coding for mannanase predicted from the partial genomic sequence of Paenibacillus woosongensis was cloned into Escherichia coli by polymerase chain reaction amplification, and completely sequenced. This mannanase gene, designated man26AT, consisted of 3,162 nucleotides encoding a polypeptide of 1,053 amino acid residues. Based on the deduced amino acid sequence, Man26AT was identified as a modular enzyme, which included a catalytic domain belonging to the glycosyl hydrolase family 26 and two carbohydrate-binding modules, CBM27 and CBM11. The amino acid sequence of Man26AT was homologous to that of several putative mannanases, with identity of 81% for P. ihumii and identity of less than 57% for other strains of Paenibacillus. A cell-free extract of recombinant E. coli carrying the man26AT gene showed maximal mannanase activity at $55^{\circ}C$ and pH 5.5. The enzyme retained above 80% of maximal activity after preincubation for 1 h at $50^{\circ}C$. Man26AT was comparably active on locust bean gum (LBG), galactomanan, and kojac glucomannan, whereas it did not exhibit activity on carboxymethylcellulose, xylan, or para-nitrophenyl-${\beta}$-mannopyranoside. The common end products liberated from mannooligosaccharides, including mannotriose, mannotetraose, mannopentaose, and mannohexaose, or LBG by Man26AT were mannose, mannobiose, and mannotriose. Mannooligosacchrides larger than mannotriose were found in enzymatic hydrolyzates of LBG and guar gum, respectively. However, Man26AT was unable to hydrolyze mannobiose. Man26AT was intracellularly degraded into at least three active proteins with different molecular masses by zymogram.

• Korean Journal of Microbiology
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• v.54 no.2
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• pp.126-135
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• 2018

The characteristics of enzyme and gene for mannanase B had been reported from Cellulosimicrobium sp. YB-43 producing some kind of mannanase. A gene coding for the enzyme, named mannanase C (ManC), was expected to be located downstream of the manB gene. The manC gene was cloned by polymerase chain reaction and sequenced completely. From this nucleotide sequence, ManC was identified to consist of 448 amino residues and contain a carbohydrate binding domain CBM2 besides a catalytic domain, which was homologous to mannanase belonging to the glycosyl hydrolase family 5. The catalytic domain of ManC showed the highest amino acid sequence similarity of 55% with the mannanases from Streptomyces sp. SirexAA-E (55.8%; 4FK9_A) and S. thermoluteus (57.6%; BAM62868). The His-tagged ManC (HtManC) lacking N-terminal signal peptide with hexahistidine at C-terminus was produced and purified from cell extract of recombinant Escherichia coli. The purified HtManC showed maximal activity at $65^{\circ}C$ and pH 7.5, with no significant change in its activity at pH range from 7.5 to 10. HtManC showed more active on konjac and locust bean gum (LBG) than guar gum and ivory nut mannan (ivory nut). Vmax and Km values of the HtManC for LBG were 68 U/mg and 0.45 mg/ml on the optimal condition, respectively. Mannobiose and mannotriose were observed on TLC as major products resulting from the HtManC hydrolysis of mannooligosacharides. In addition, mannobiose and mannose were commonly detected as the hydrolyzed products of LBG, konjac and ivory nut.

• Food Science and Biotechnology
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• v.15 no.5
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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.

• Korean Journal of Microbiology
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• v.51 no.3
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• pp.263-270
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• 2015

A bacterial strain producing extracellular mannanases was isolated from soil of chestnut tree farm located in Gongju city of Korea by enrichment culture using Avicel as a carbon source. 16S rDNA sequence of the isolate YB-43 was highly homologous to those of genus Cellulosimicrobium strains with sequence similarities of above 99.6%. Mannanase productivity was significantly increased when the Cellulosimicrobium sp. YB-43 was grown in the presence of locust bean gum (LBG) or konjac. The mannanases were partially purified to be mannanase A (ManA) and mannanase C (ManC) by DEAE-Sepharose column and Q-Sepharose column chromatography from the culture filtrate of Cellulosimicrobium sp. YB-43 grown in LB medium supplemented with 0.7% LBG for 24 h. The partially purified ManA showed the highest activity at $55^{\circ}C$ and pH 6.5, while ManC activity was optimal at $65^{\circ}C$ and pH 7.5. ManA was stable up to $40^{\circ}C$ for 1 h, but ManC activity decreased significantly even after 1 h at $20^{\circ}C$. ManA and ManC showed difference from each other according to their substrate specificities and predominant products resulting from the mannanase hydrolysis for mannooligosaccharides. As a result, Cellulosimicrobium sp. YB-43 was found to produce two different kinds of mannanases.

• Microbiology and Biotechnology Letters
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• v.31 no.3
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• pp.277-283
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• 2003

A bacterium producing the extracellular mannanase was isolated from Korean soybean paste. The isolate WL-7 has been identified as Bacillus subtiis on the basis on its 16S rRNA sequence, fatty acid composition, morphology and biochemical properties. The mannanase of culture supernatant was the most active around $55^{\circ}C$ and pH $6.0^{\circ}C$, and retained 90% of its maximum activity at range of pH 5.0∼7.5 and $50∼60^{\circ}C$. The additional carbohydrates including lactose, $\alpha$-cellulose, avicel, locust bean gum (LBG), wheat bran and konjak increased dramatically the mannanase productivity of strain WL-7. Especially, the maximum mannanase productivity was reached to 224 U/ml in LB medium supplemented with both 0.5% LBG and 0.5% konjak, which was approximately 200-folds more than that in LB medium. It was suggested that the increase of mannanase production was owing to induction of mannanase biosynthesis by both LBG and konjak hydrolysates transported following initial hydrolysis by extracellular mannanase during the cell growth.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.4
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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 Life Science
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• v.26 no.10
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• pp.1113-1120
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• 2016

Two bacterial strains producing extracellular man nanase were isolated from doenjang, a traditionally fermented soybean paste in Korea. The isolates, WL-6 and WL-11, were identified as Bacillus subtiis on the basis of their 16S rRNA gene sequences, morphological, and biochemical properties. Two genes encoding the mannanase of both B. subtilis WL-6 and B. subtilis WL-11 were each cloned into Escherichia coli, and their nucleotide sequences were determined. Both mannanase genes consisted of 1,086 nucleotides, encoding polypeptides of 362 amino acid residues. The deduced amino acid sequences of the two WL-6 and WL-11 mannanases, designated Man6 and Man11, respectively, differed from each other by eight amino acid residues, and they were highly homologous to those of mannanases belonging to the glycosyl hydrolase family 26. The 26 amino acid stretch in the N-terminus of Man6 and Man11 was a predicted signal peptide. Both Man6 and Man11 were localized at the level of 94–95% in an intracellular fraction of recombinant E. coli cells. The enzymes hydrolyzed both locust bean gum and mannooligosaccharides, including mannotriose, mannotetraose, mannopentaose, and mannohexaose, forming mannobiose and mannotriose as predominant products. The optimal reaction conditions were 55°C and pH 6.0 for Man6, and 60°C and pH 5.5 for Man11. Man11 was more stable than Man6 at high temperatures.

• Microbiology and Biotechnology Letters
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• v.26 no.3
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• pp.232-237
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• 1998

A bacterial strain producing high levels of an extracellular ${eta}$-mannanase and intracellular ${eta}$-mannosidase and ${alpha}$-galactosidase was isolated from soil. The strain isolated was identified as a strain of Sporolactobacillus sp. and designated as Sporolactobacillus sp. M20l. Synthesis of ${eta}$-mannanase by Sporolactobacillus sp. M20l was induced by sucrose, maltose, or locust bean gum. The highest induction rate was obtained with 2% locust bean gum added to the culture medium as a sole carbon source. On the other hand, induction of ${eta}$-mannosidase was observed only with locust bean gum. The optimal media for the enzyme production were established as follows: for ${eta}$-mannanase; 2% locust bean gum, 0.5% peptone, 0.2% KH$_2$PO$_4$, 80 mg/l MgSO$_4$, and 8 mg/l ZnSO$_4$ (pH 6.0), and for ${eta}$-mannosidase; 2% locust bean gum, 0.5% yeast extract, 0.2% KH$_2$PO$_4$, 80 mg/l MgSO$_4$, and 8 mg/l ZnSO$_4$ (pH 5.0). The optimal culture temperatures for production of ${eta}$-mannanase and ${eta}$-mannosidase were found to be 37$^{\circ}C$ and 3$0^{\circ}C$, respectively. Under the optimal culture conditions, the production of ${eta}$-mannanase and ${eta}$-mannosidase reached the highest levels of 10.6 units/ml and 1.35 units/ml after 30 h and 24 h cultivation, respectively.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.4
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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.