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( Gwi Gun Park ) 한국산업식품공학회 2018 산업 식품공학 Vol.22 No.3
O-D-mannopyranosyl-(1→4)-O-D-glucopyranosyl-(1→4)-D-Mannopyranose (MGM) was prepared via the enzymatic hydrolysis of konjac glucomannan and the subsequent elimination of monosaccharides from the resultant hydrolysate using yeast. The enzyme system hydrolyzed konjac glucomannan and produced monosaccharides and MGM without other oligomers at the 48 h reaction. Konjac 20 g was hydrolyzed at 60℃ and a pH 6.0 for 48 h with 200 mL crude enzyme solution from Xylogone sphaerospora. By eliminating monosaccharides from the hydrolysis products with yeast (Candida guilliermondii), 3.8 g of crystalline MGM was obtained, without the use of chromatographic techniques. After 48 h of yeast cultivation, the total sugar content fell from 5.2% to 3.7%, while the average degree of polymerization (D.P.) rose from 2.6 to 3.3.
Gwi Gun Park 한국응용생명화학회 2008 Applied Biological Chemistry (Appl Biol Chem) Vol.51 No.6
Three kinds of oligosaccharides were obtained from the hydrolysate of brown copra meal galactomannan by a purified extracellular β-mannanase from Trichoderma sp. These oligosaccharides were identified as Man-Man, Gal2Man3 (62-mono-O-α-D-galactopyranosyl-4-O-β-D-mannotriose), and Gal2Man6 (62-mono- O-α-D-galactopyranosyl-4-O-β-D-mannohexaose), where Gal- and Manrepresent α-1,6-D-galactosidic and β-1,4-mannosidic linkages, respectively. The mode of action of β-mannanase on brown copra meal galactomannan is described on the basis of the structure of these oligosaccharides. Three kinds of oligosaccharides were obtained from the hydrolysate of brown copra meal galactomannan by a purified extracellular β-mannanase from Trichoderma sp. These oligosaccharides were identified as Man-Man, Gal2Man3 (62-mono-O-α-D-galactopyranosyl-4-O-β-D-mannotriose), and Gal2Man6 (62-mono- O-α-D-galactopyranosyl-4-O-β-D-mannohexaose), where Gal- and Manrepresent α-1,6-D-galactosidic and β-1,4-mannosidic linkages, respectively. The mode of action of β-mannanase on brown copra meal galactomannan is described on the basis of the structure of these oligosaccharides.
Gwi-Gun Park 한국식품영양과학회 2000 Preventive Nutrition and Food Science Vol.5 No.4
β-1,4-Mannotriose was prepared by the enzymatic hydrolysis of white copra meal (WCM) and the subsequent elimination of monosaccharides from the resultant hydrolysate with a yeast. The enzyme system from sunflower seed hydrolyzed WCM and produced monosaccharides and β-1,4-mannotriose without other oligomers at the final stage of the reaction. WCM (50 g) was hydrolyzed at 50℃ and pH 4.5 for 24 hr with a crude enzyme solution (500 mL) from sunflower seed. By the elimination of monosaccharides from the hydrolysis products with a yeast (Candida glaebosa), 8.1 g of crystalline mannotriose was otained without the use of chromatographic techniques. After 48hr of yeast cultivation, the total sugar content decreased from 4.6% to 3.5%, whereas the average degree of polymerization increased from 2.3 to 3.1.
Bacillus sp.유래 β - Mannanase의 정제 및 Chromatography에 의한 Xanthan Gum 가수분해물의 분리
박귀근(Gwi-Gun Park) 한국식품영양과학회 2003 한국식품영양과학회지 Vol.32 No.4
DEAE Sepahcel ion exchange chromatography(2.5×42㎝)에 의해 Bacillus sp. 유래 β-Mannanase정제를 수행하였다. 정제효소의 비활성은 17.41 units/㎎로서 정제배율은 84.74배를 나타내었다. Carbon column chromatography를 이용하여 0~50%의 ethanol gradient법으로 xanthan gum의 가수분해물을 분리한 결과 fraction number 40~45 및 50~60사이에서 broad한 2개 peak의 가수분해물 pattern을 나타내었다. 가수분해물의 분리도를 확인하기 위하여 TLC를 수행한 결과 fraction No. 40~44에서는 Rf value상 중합도 5에 해당하는 가수분해물이 주축을 이루고 있는 반면 fraction No. 50~55에서는 중합도 7의 가수분해물이 주축을 이루고 있음을 확인할 수 있었다. 중합도별 가수분해물의 분리도를 높이기 위해 2차 Sephadex G-25 column chromatography를 수행한 결과 fraction No. 12~15에서 중합도 7의 올리고당과 fraction No. 77~80에서 중합도 5의 가수분 해물을 분리할 수 있었고, 가수분해물의 분리도를 확인하기 위해 2차 TLC를 수행한 결과 fraction No. 12~15에서는 중합도 7이 주축을 이루고 있으나 일부 소량의 고중합도 가수 분해물이 공존하고 있는 것으로 사료되며, fraction No. 77~80에서는 분리능이 높게 중합도 5의 가수분해물이 분리되었다. 이와 같이 분리된 2개의 fractions은 FACE법에 의해 Homo type가수분해물로 동정되었다. A β-mannanase of Bacillus sp. was purified by DEAE Sephacel ion exchange column chromatography. The specific activity of the purified enzyme was 17.41 units/㎎ protein, representing an 84.74-folds purification of the original crude extract. For the separation of two types of hydrolysates by the action of purified β-mannanase, carbon column chromatography, sephadex G-25 column chromatography and thin layer chromatography were accomplished. Main hydrolysates were D.P value 5 and 7 containing of low D.P values. By the method of FACE (Fluorophore Assisted Carbohydrate Electrophoresis), two types of hydrolysates were identified to homo type.
Production of Mannooligosaccharides by the Penicillium purpurogenum Mannanase
Park, Gwi-Gun The Korean Society of Food Science and Nutrition 1994 한국식품영양과학회지 Vol.23 No.3
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.