• Journal of the Korean Society of Food Science and Nutrition
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• v.23 no.3
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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 Applied Biological Chemistry
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• v.51 no.6
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• pp.272-276
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• 2008

Purification and some properties of Xylogone sphaerospora ${\beta}$-mannanase were reprevious previous paper. Locust bean gum galactomannan was hydrolyzed by the purified ${\beta}$-mannanase, and then the hydrolysates was separated by activated carbon column chromatography. The main hydrolysates were composed of D.P. (Degree of Polymerization) 4 and 6 galactosyl mannooligosaccharides. For elucidate the structure of D.P 4 and 6 galactosyl mannooligosaccharides, sequential enzymatic action was performed. D.P 4 and 6 were identified as ${Gal^2}{Man_3}\;(6^2-mono-O-{\alpha}-D-galactopyranosyl-4-O-{\beta}-D-mannotriose)$ and ${Gal^2}{Man_5}\;(6^2-mono-O-{\alpha}-D-galacto- pyranosyl-4-O-{\beta}-D-mannopentaose)$. To investigate the effects of locust bean gum galactosyl mannooligosaccharides on in vitro growth of Bifidobacterium longum, B. bifidum, B. infantis, B. adolescentis, B. animalis, B. auglutum and B. breve. Bifidobacterium spp. were cultivated individually on the modified-MRS medium containing carbon source such as D.P. 4 and D.P. 6 galactosyl mannooligosaccharides, respectively. B. longum and B. bifidum grew up to-fold and 6.6-fold more effectively by the treatment of D.P. 6 galactosyl mannooligosaccharides, compared to those of standard MRS medium. Especially, D.P. 6 was more effective than D.P. 4 galactosyl mannooligosaccharide on the growth of Bifidobacterium spp.

• Applied Biological Chemistry
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• v.48 no.4
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• pp.364-369
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• 2005

This study was performed to elucidate substrate specificity to the locust bean gum galactomannan by Trichoderma harzianum ${\beta}-mannanase$. The medium composition for enzyme production were determined 3% cellulose, 3% corn steep liquor, 1% $KH_2PO_4$, 0.2% $(NH_4){_2}SO_4$, and incubated for 115 hr at $28^{\circ}C$. The ${\beta}-mannanase$ exhibited maximum activity at pH 4.5 and $60^{\circ}C$. Locust bean gum galactomannan was hydrolyzed by the ${\beta}-mannanase$, and then hydrolysates separated by activated carbon column chromatography. The main hydrolysates were composed of D.P 4 and 7 galactosyl mannooligosaccharides by TLC. For the elucidate the structure of D.P 4 and 7 oligosaccharides, methylation analysis was performed. D.P 4 and 7 were identified as M-M-M-M and M-M-M-M-M (G- and M-represent ${\alpha-1,6-D-galactosidic\;and\;{\beta}-1,4-mannosidic$ linkages, respectively). //G-G To investigate the effects of locust bean gum galactosyl mannooligosaccharides on the in vitro growth of B. longum, B. bifidum, B. infantis, and B. breve, Bifidobacterium spp. were cultivated individually on the modified-MRS medium containing carbon source such as D.P 4 and 7 galactosyl mannooligosaccharides, respectively. B. longum grew up 3.4-fold and 4.3-fold more effectively by the replacement of D.P 4 and 7 galactosyl mannooligosaccharides as the carbon source in a comparasion of standard MRS.

• Korean Journal of Microbiology
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• v.50 no.2
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• pp.158-163
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• 2014

In the acidic LB plate, a bacterial strain was isolated from homemade soybean paste as a producer of the extracellular mannanase. The isolate YB-1402, which was a Gram-positive rod-shaped bacterium with spore, has been identified as Bacillus amyloliquefaciens on the basis of its 16S rDNA sequence and biochemical properties. Maximum mannanase productivity of the isolate YB-1402 was reached approximately 150 U/ml in LB broth supplemented with konjac (3.0%). The molecular mass of YB-1402 mannanase was estimated to approximately 38.0 kDa by zymogram of the culture filtrate on SDS-PAGE. The mannanase of culture filtrate was the most active at $55^{\circ}C$ and pH 5.5. The mannanase activity was completely maintained after pre-incubation at pH 3.0 to 10.0 for 1 h. The predominant products resulting from the mannanase hydrolysis were mannose, mannobiose and mannotriose for LBG or mannooligosaccharides. The enzyme could hydrolyze mannooligosaccharides larger than mannobiose.

• Journal of Applied Biological Chemistry
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• v.53 no.2
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• pp.71-76
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• 2010

A gene encoding the mannanase of Bacillus licheniformis WL-12, which had been isolated from Korean soybean paste, was cloned into Escherichia coli and nucleotide sequence of the mannanase gene was subsequently determined. The mannanase gene consisted of 1,080 nucleotides encoding a polypeptide of 360 amino acid residues. The deduced amino acid sequence was identical to that of putative mannanase from B. liceniformis DSM13 belonging to GH family 26. The mannanase was partially purified from cell-free extract of the recombinant Escherichia coli carrying a WL-12 mannanase gene by ammonium sulfate fractionation and DEAE-Sepharose column chromatography. Optimal conditions for the partially purified enzyme occurred at pH 6.0 and $65^{\circ}C$. The enzyme showed higher activity on locust bean gum (LBG) galactomannan and konjac glucomannan than on guar gum galactomannan. The predominant products resulting from the mannanase hydrolysis were mannose, mannobiose and mannotriose for LBG or mannooligosaccharides. The enzyme could hydrolyze mannooligosaccharides larger than mannobiose.

• Korean Journal of Microbiology
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• v.52 no.3
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• pp.336-343
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• 2016

A mannanase gene was cloned into Escherichia coli from Cellulosimicrobium sp. YB-43, which had been found to produce two kinds of mannanase, and sequenced completely. This mannanase gene, designated manB, consisted of 1,284 nucleotides encoding a polypeptide of 427 amino acid residues. Based on the deduced amino acid sequence, the ManB was identified to be a modular enzyme including two carbohydrate binding domains besides the catalytic domain, which was highly homologous to mannanases belonging to the glycosyl hydrolase family 5. The N-terminal amino acid sequence of ManB, purified from a cell-free extract of the recombinant E. coli carrying a Cellulosimicrobium sp. YB-43 manB gene, has been determined as QGASAASDG, which was correctly corresponding to signal peptide predicted by SignalP4.1 server for Gram-negative bacteria. The purified ManB had a pH optimum for its activity at pH 6.5~7.0 and a temperature optimum at $55^{\circ}C$. The enzyme was active on locust bean gum (LBG), konjac and guar gum, while it did not exhibit activity towards carboxymethylcellulose, xylan, starch, and para-nitrophenyl-${\beta}$-mannopyranoside. The activity of enzyme was inhibited very slightly by $Mg^{2+}$, $K^+$, and $Na^+$, and significantly inhibited by $Cu^{2+}$, $Zn^{2+}$, $Mn^{2+}$, and SDS. The enzyme could hydrolyze mannooligosaccharides larger than mannobiose, which was the most predominant product resulting from the ManB hydrolysis for mannooligosaccharides and LBG.

• Applied Biological Chemistry
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• v.47 no.4
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• pp.379-383
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• 2004

For the elucidation of substrate specificity to the brown copra meal by Bacillus sp. ${\beta}-mannanase.$, the enzymatic hydrolysate after 24 hr of reaction was heated in a boiling water bath for 10 min, and then centrifuged to remove the insoluble materials from hydrolysates. The major hydrolysates composed of D.P 5 and 7 galactosyl mannooligosaccharides. For the separate of galactosyl mannooligosaccharides, the supernatant solution of 150 ml was put on a first activated carbon column. The column was then washed with 5 l of water to remove mannose and salts. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol, at the flow rate of 250 ml per hour. The sugar composition in each fraction tubes was examined by TLC and FACE analysis. The combined fraction from F3 was concentrated to 30 ml by vacuum evaporator. Then put on a second activated carbon column. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol (total volume: 5 l), at the flow rate of 250 ml per hour. The eluent was collected in 8 ml fraction tubes, and the total sugar concentration was measured by method of phenol-sulfuric acid. The major component of F2 separated by 2nd activated carbon column chromatography were identified $Gal^3Man_4(6^3-mono-{\alpha}-D-galactopyranosyl-{\beta}-mannotetraose)$. To investigate the effects of brown copra meal galactomannooligosaccharides on growth of Bifidobacterium longum, B. bifidum were cultivated individually on the modified-MRS medium containing carbon source such as $Gal^3Man_4$, compared to those of standard MRS medium.

• Microbiology and Biotechnology Letters
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• v.42 no.2
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• pp.99-105
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• 2014

A Bacillus strain capable of hydrolyzing locust bean gum was isolated as a producer of extracellular mannanase by way of an enrichment culture in an acidic medium from homemade soybean pastes. The isolate YB-1401 showed a biochemical identity of 61.1% with Brevibacillus laterosporus, while the nucleotide sequence of its 16S rDNA had the highest similarity with that of Bacillus amyloliquefaciens. The mannanase productivity of the Bacillus sp. YB-1401 was drastically increased by mannans. Particularly, maximum mannanase productivity was reached at approximately 265 U/ml in LB medium supplemented with konjac glucomannan (4.0%). The mannanase was the most active at $55^{\circ}C$ and pH 5.5. Mannanase activity was completely maintained after pre-incubation at pH 3.5 to 11.0 for 1 h. The predominant products resulting from the mannanase hydrolysis were mannobiose and mannotriose for LBG, guar gum or mannooligosaccharides. A small amount of mannose was also detected in the hydrolyzates.

• 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.

• Applied Biological Chemistry
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• v.47 no.4
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• pp.384-389
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• 2004

The purified ${\beta}-mannanase$ hydrolyzed various gums to mannose, ${\beta}-1,4-mannobiose$, $Gal^3Man_4$, and D.P 7 of galactosyl mannooligosaccharide, and isolated from the enzymatic hydrolysate for 24 hrs reaction by activated carbon column chromatography and Sephadex G-25 column chromatography. For the elucidate of antioxidant action of ${\beta}-1,4-mannobiose$, $Gal^3Man_4$ and DP 7 of galactosyl mannooligosaccharide and urea derivatives, coloration, reducing power, antioxidant activity and DPPH test were accomplished. The coloration was high at reaction mixture of ${\beta}-1,4-mannobiose$, $Gal^3Man_4$ D.P 7 and urea. TLC of reaction mixture of ${\beta}-1,4-mannobiose$, $Gal^3Man_4$ D.P 7 and ureas showed new reaction products, respectively. but except reaction mixture of ${\beta}-1,4-mannobiose$ and urea. The reducing power was high at reaction mixture of ${\beta}-1,4-mannobiose$, $Gal^3Man_4$ D.P 7 and phenylthiourea. The reaction mixture of ${\beta}-1,4-mannobiose$, $Gal^3Man_4$ D.P 7 and thiourea showed similar radical scavenging activities on DPPH to activity of AsA. The reaction mixture of ${\beta}-1,4-mannobiose$, $Gal^3Man_4$ D.P 7 and thiourea, phenythiolurea shown strong antioxidative activites on the oxidation of linoneic acid.