• Journal of Microbiology and Biotechnology
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• v.2 no.3
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• pp.189-196
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• 1992

A $\alpha$-l, 4-D-glucan maltohydrolase $(\beta$-amylase), secreted by the mesophilic aerobic bacterium Bacillus polymyxa No.26, was purified and characterized. The enzyme production was increased after a logarithmic phase of bacterial growth and paralleled with the onset of bacterial sporulation. By applying anion exchange chromatography and gel filtration the enzyme was purified 16.7-fold and had a specific activity of 285.7 units/mg. Two enzyme activities were eluted on a column of DEAE-Sephadex chromatography, and they were designated as E-I for a major enzyme peak and E-II for a minor peak. Of them, E-I enzyme peak was further purified by using gel chromatography. The molecular mass of this enzyme was determined to be 64, 000 daltons and consisted of a single subunit, showing an isoelectric point of 8.9. The enzyme was able to attack specifically the $\alpha$-l, 4-glycosidic linkages in soluble starch and caused its complete hydrolysis to maltose and $\beta$-limited dextrin. This amylolytic enzyme displayed a temperature optimum at $45^\circ{C}$ and a pH optimum at 7.0. The amino acid composition of the purified enzyme was quite similar to the other bacterial $\beta$-amylases reported. Surprisingly, the purified enzyme from this aerobe only exhibited hydrolytic activity on soluble starch, not on starch granules. The degradation of from starch by $\beta$-amylase was greatly stimulated by pullulanase addition. These results differentiated from other $\beta$-amylases reported. Based on a previous result that showed the enzyme system involves in effective degradation of raw starch granules, this result strongly suggested that the purified enzyme (E-I) can be a synergistic part of starch granule-digestion and E-II plays a crucial role in digestion of starch granules.

• Journal of Life Science
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• v.11 no.5
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• pp.393-399
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• 2001

In order to improve functionality of kochujang which is one of the traditional foods in Korea, sea tangle powder(2, 4, 6 and 8% sea tangle powder on the glutinous rice weight basis) was added to the raw material of kochujang and then investigated the bacterial counts and enzyme activities with control kochujang during the fermentation at 3$0^{\circ}C$ for 120 days. Bacterial count was about 10$^4$ cfu/g at initial stage of fermentation and then maintained 10$^{8}$ cfu/g after 60 days of fermentation. $\alpha$-amylase activity was gradually reduced during fermentation periods, so the activity was lost almost at late of fermentation $\beta$-amylase activity was rapid increased until 30 days of fermentation and the rapid decreased at 60 days of fermentation and after 90 days was slightly decreased. Activities of acidic protease and neutral protease were increased until 30 days of fermentation and then these were shown irregularities decreased.

• Journal of Microbiology and Biotechnology
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• v.27 no.5
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• pp.896-908
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• 2017

In this study, a total of 58 different kinds of nuruk (a traditional Korean fermentation starter) were prepared, including 46 kinds of restored nuruk from ancient documents. Each nuruk was evaluated by analysis of its saccharification power, and the enzyme activities of glucoamylase, ${\alpha}$-amylase, ${\beta}$-amylase, protease, and ${\beta}$-glucanase. The range of saccharification power (sp) of the restored nuruk ranged between 85 and 565 sp. The diastatic enzymes, ${\alpha}$-amylase, ${\beta}$-amylase, and glucoamylase, were significantly correlated to the saccharification power value; conversely, ${\beta}$-glucanase and protease did not have a correlative relationship with saccarification power. In addition, their brewing properties on chemical and organoleptic aspects of traditional alcoholic beverage production were compared. Each raw and supplementary material contained in nuruk showed its own unique characteristics on Korean alcoholic beverage brewing. For the first time, in this study, the traditional Korean nuruk types mentioned in ancient documents were restored using modernized production methods, and also characterized based on their brewing properties. Our results could be utilized as a basis for further study of traditional alcoholic beverages and their valuable microorganisms.

• Journal of Microbiology
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• v.42 no.4
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• pp.319-327
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• 2004

An additional amylase, besides the typical $\alpha-amylase,$ was detected for the first time in the cytoplasm of B. subtilis SUH4-2, an isolate from Korean soil. The corresponding gene (bbmA) encoded a malto­genic amylase (MAase) and its sequence was almost identical to the yvdF gene of B. subtilis 168, whose function was unknown. Southern blot analysis using bbmA as the probe indicated that this gene was ubiquitous among various B. subtilis strains. In an effort to understand the physiological function of the bbmA gene in B. subtilis, the expression pattern of the gene was monitored by measuring the $\beta-galactosidase$ activity produced from the bbmA promoter fused to the amino terminus of the lacZ struc­tural gene, which was then integrated into the amyE locus on the B. subtilis 168 chromosome. The pro­moter was induced during the mid-log phase and fully expressed at the early stationary phase in defined media containing $\beta--cyclodextrin\;(\beta-CD),$ maltose, or starch. On the other hand, it was kept repressed in the presence of glucose, fructose, sucrose, or glycerol, suggesting that catabolite repression might be involved in the expression of the gene. Production of the $\beta-CD$ hydrolyzing activity was impaired by the spo0A mutation in B. subtilis 168, indicating the involvement of an additional regu­latory system exerting control on the promoter. Inactivation of yvdF resulted in a significant decrease of the $\beta-CD$ hydrolyzing activity, if not all. This result implied the presence of an additional enzyme(s) that is capable of hydrolyzing $\beta-CD$ in B. subtilis 168. Based on the results, MAase encoded by bbmA is likely to be involved in maltose and $\beta-CD$ utilization when other sugars, which are readily usable as an energy source, are not available during the stationary phase.

• The Korean Journal of Food And Nutrition
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• v.10 no.1
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• pp.92-96
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• 1997

A Korean commercial sweet rice drink "Sikhye" showed sucrose, fructose, glucose, maltose, limit dextrin and various size of maltooligosaccharides in HPLC and TLC analysis. Commercial Sikhye was found to contain 0.09% of limit dextrin and 0.2% of rice residue. Limit dextrin in commercial Sikhye showed both signal of $\alpha$-1,4- and $\alpha$-1,6-glucosidic linkage with its estimation ratio of 15:1 by 1H-NMR analysis. This limit dextrin was hydrolyzed to produce various size of maltooligosaccarides with more longer chain than that of traditional Sikhye by pullulanase. Limit dextrin was digested wit enzymes(30units/ml) of $\alpha$-amylase, $\alpha$-glucosidase and glucoamylase from Aspergillus awamori, sweet potato $\beta$-amylase and human salivary $\alpha$-amylase at 37$^{\circ}C$ for 1 hour, respectively. Hydrolysis rates of these amylases on it were higher than in case of traditional sikhye. $\alpha$-Glucosidase plus human salivary $\alpha$-amylase hydrolyzed it to 61.3%. Hydrolysis rates of these amylases on rice residue were lower than that of traditional Sikye. These results suggest that limit dextrin in commercial Sikhye is less effective than isomaltooligosaccharides in traditional Sikhye as a growth factor for Bifidobacterium while rice residue in commercial Sikhye is more effective than that in traditional Sikhye as dietary fiber.ary fiber.

• KSBB Journal
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• v.22 no.2
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• pp.114-118
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• 2007

In this study, an amylase-producing fungus, strain 15 was isolated from soil in order to saccharify food wastes with cellulolytic and amylolytic enzymes. The amylase production cultures were performed in Mandel's medium with 1% rice straw and 1% paper wastes as carbon sources. The strain produced various cellulolytic (FPase 0.25, xylanase 20.09, CMCase 3.15 U/mL-supernatant) and amylolytic ($\alpha$-amylase 1.20, gluco-amylase 0.70, $\beta$-amylase 2.40 U/mL-supernatant) enzymes in Mandel's medium. In 10 L jar fermenter, maximum amylase and FPase activities, 3.25 and 0.23 U/mL, were obtained when the culture was grown at 30$^{\circ}C$, 200 rpm and 0.6 vvm for 3 days. In 100 mL flask level and 10 L jar fermenter, amylase produced by the strain 15 showed similar cellulolytic and amylolytic enzyme activities with Trichoderma inhamatum KSJ1 isolated from rotten woods by previous researcher. The ability of saccharification to food wastes also showed similar degree. However, the isolate 15 appeared to be yellowish in YMEA plate comparing to Trichoderma inhamatum KSJ1 in greenish.

• The Korean Journal of Mycology
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• v.40 no.3
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• pp.152-158
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• 2012

The lignocellulytic enzymes including a-amylase (EC 3.2.1.1), lignin peroxidase (EC 1.11.1.14), laccase (EC 1.10.3.2), xylanase (EC 3.2.1.8), ${\beta}$-xylosidase (EC 3.2.1.37), ${\beta}$-glucosidase (EC 3.2.1.21) and cellulase (EC 3.2.1.4) were extracted from spent mushroom compost (SMC) of Pleurotus eryngii. Different extraction buffers and conditions were tested for optimal recovery of the enzymes. The optimum extraction was shaking incubation (200 rpm) for 2 h at $4^{\circ}C$. ${\alpha}$-Amylase was extracted with the productivity range from 1.20 to 1.6 Unit/SMC g. Cellulase was recovered with the productivity range from 2.10 to 2.80 U/gf. ${\beta}$-glucosidase and ${\beta}$-xylosidase productivities showed lowest recovery producing 0.1 U/g and 0.02 U/g, respectively. The P. eryngii SMCs collected from three different mushroom farms showed different recovery on laccase and xylanse, cellulase. Furthermore, the water extracted SMC was compared to commercial enzymes for its industrial application in decolorization and cellulase activity.

• Journal of Plant Biology
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• v.31 no.4
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• pp.249-258
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• 1988

Changes in starch content and activities of ADPG- and UDPG-starch synthase and $\alpha$- and, $\beta$-amylase were studied in order to investigate effects of gibberellic aicd and abscisic acid on endogenous starch content during shoot differentiation and protocorm propagation in Cymbidium spp. (Jungfrau) protocorm. Shoot differentiation was promoted during the degradation of endogenous starch and protocorn propagation was promoted during starch accumulation in protocorm. The activities of ADPG- and, UDPG-starch synthase and $\alpha$- and $\beta$-amylase seemed to be related with starch content. Shoot differentiation and protocorm propagation were slightly inhibited in protocorm explants treated with 100$\mu$M gibberellic acid. The explants treated with 10$\mu$M abscisic acid lost the capacity for shoot differentiation and protocorm propagation, and that could not be overcome by 100$\mu$M gibberellic acid added to culture medium. Starch content fluctuated as the control even after 10$\mu$M abscisic acid. None the less, the treatment completely inhibited shoot differentiation and protocorm propagation.

• Microbiology and Biotechnology Letters
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• v.25 no.2
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• pp.109-114
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• 1997

The Oomycete Saprolegnia ferax produces an extracellular $\beta$-amylase, Maximum enzyme yield was attained after 7 days of growth in YNB starch medium (pH 6.5) at 25$\circ$C. The amylase was pu- rified 24-fold by ultrafitration, HPLC DEAE column and HPLC gel filtration. The purfied enzyme was a monomeric glycoprotein with a molecular weight of about 44,000 dalton. The pH and temperature optima were 6.5 and 50$\circ$C, respectively. The enzyme was fairly stable up to 50$\circ$C and at acidic pH region (pH 4.0-7.0). The apparent Km and Vmax values of the enzyme against soluble starch were 0.77 mg/ml and 2,174 $\mu$moles/mg protein, respectively. Amino acid analysis indicated that the enzyme was enriched in alanine, glycine, leucine and acidic amino acid. Starch hydrolysis with the enzyme released maltose but not glucose, whereas maltotriose, Schardinger dextrin ($\alpha$-cyclodextrin) and pullulan were not hydrolysed by the enzyme. The enzyme was inhibited by Schardinger dextrin, p-chloromercuribenzoate(PCMB), CU$^{2+}$' and Hg$^{2+}$. Inhibition of the enzyme by PCMB could be reversed by the addition of cysteine and mercaptoethanol.

• Korean Journal of Food Science and Technology
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• v.29 no.2
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• pp.284-291
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• 1997

Microwave vacuum heating method (2450 MHz) was used for a low intensity of heat treatments. High vacuum under the microwave heating could bring low temperature condition. Inactivation of ${\alpha}-amylase,\;{\beta}-amylase$, glucoamylase and peroxidase by microwave vacuum heating were investigated at 60-$80^{\circ}C$. It was compared with conventional heating. The heating condition of microwave vacuum heating was confirmed by the destruction of ascorbic acid. When thermal inactivations of the enzymes by microwave vacuum heating were determined, it was less effective than that of conventional method at the initial stage of heating. It was due to a lag time of microwave heating. However, the heating time for complete inactivation of the enzymes by microwave vacuum heating could be reduced comparing with that of conventional heating. Optimum conditions for inactivation of the enzymes could be obtained by microwave vacuum heating.