• Journal of Microbiology and Biotechnology
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• v.19 no.11
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• pp.1306-1318
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• 2009

The filamentous ascomycete Sclerotinia sclerotiorum is well known for its ability to produce a large variety of hydrolytic enzymes. Two $\alpha$-amylases ScAmy54 and ScAmy43 predicted to play an important role in starch degradation were showed to produce specific oligosaccharides essentially maltotriose that have a considerable commercial interest. Primary structure of the two enzymes was established by N-terminal sequencing, MALDI-TOF masse spectrometry and cDNA cloning. The two proteins have the same N-terminal catalytic domain and ScAmy43 derived from ScAmy54 by truncation of 96 amino acids at the carboxyl-terminal region. Data of genomic analysis suggested that the two enzymes originated from the same $\alpha$-amylase gene and that truncation of ScAmy54 to ScAmy43 occurred probably during S. sclerotiorum cultivation. The structural gene of Scamy54 consisted of 9 exons and 8 introns, containing a single 1,500-bp open reading frame encoding 499 amino acids including a signal peptide of 21 residues. ScAmy54 exhibited high amino acid homology with other liquefying fungal $\alpha$-amylases essentially in the four conserved regions and in the putative catalytic triad. A 3D structure model of ScAmy54 and ScAmy43 was built using the 3-D structure of 2guy from A. niger as template. ScAmy54 is composed by three domains A, B, and C, including the well-known $(\beta/\alpha)_8$ barrel motif in domain A, have a typical structure of $\alpha$-amylase family, whereas ScAmy43 contained only tow domains A and B is the first fungal $\alpha$-amylase described until now with the smallest catalytic domain.

• Proceedings of the Microbiological Society of Korea Conference
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• 2003.05a
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• pp.122-124
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• 2003

Cyclodextrin glucanotransferase (CGTase) and endoxylanase genes of Bacillus sp. were subcloned down-stream of yeast ADH1 promoter and expressed in S. cerevisiae. Most of the CGTase and endoxylanase expressed were detected in the extracellular medium with activity of 0.6 and 7-8 unit/ml, respectively. The recombinant enzymes were secreted as N-linked-glycosylated forms, resulting an enhanced thermal stability. CGTase predominantly produced $\alpha-cyclodextrin$ from starch and endoxylanase produced xylobiose and xylotriose from xylan.

• Mycobiology
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• v.37 no.4
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• pp.286-294
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• 2009

Heteropolysaccharides isolated from liquid cultures of nine Tremella species contained 0.3 to 1.2% protein, 2.7 to 5% ash, 0.9 to 3.4% acetyl groups, 76.5 to 84.2% carbohydrates and trace amounts of starch. The polysaccharides in aqueous solution were slightly acidic (pH 5.1 to 5.6). They consisted of the following monomeric sugars: fucose, ribose, xylose, arabinose, mannose, galactose, glucose and glucuronic acid. The backbones of the polysaccharide structures consisted of $\alpha$-(1$\rightarrow$3)-links while the side chains were $\beta$-linked.

• Applied Biological Chemistry
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• v.42 no.4
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• pp.324-329
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• 1999

Barley malts were prepared at 15, 18 and $21^{\circ}C$ for $3{\sim}6$ days, and assayed for ${\beta}-glucanase$, ${\alpha}-amylase$ and ${\beta}-amylase$ activities. ${\beta}-Glucanase$ activity increased markedly during earley germination and reached maximum at the 6th day of germination. ${\beta}-Glucanase$ activity in six-rowed barley malt was much higher than that in two-rowed malt. ${\beta}-Glucanase$ activity was associated with reduction in ${\beta}-glucan$ content during germination. ${\beta}-amylase$ activity was also considerably higher in two-rowed barley, and increased continuously during 6-day germination. ${\beta}-Amylase$ activity was the lowest at $15^{\circ}C$, the highest at $18^{\circ}C$, and intermediate at $21^{\circ}C$ of germination temperature. Considerable amount of ${\beta}-amylase$ was detected in ungerminated raw barley, and this enzymatic activity tended to increase during 6-day germination. Diastatic power, measure of starch-saccharifying enzyme, in six-rowed malt was $1.4{\sim}1.6$ fold higher than in two-rowed malt. Germination at $18^{\circ}C$ for $5{\sim}6$ days was suggested to be the optimum condition for manufacturing good quality malts, in terms of enhanced starch-degrading enzymatic activity.

• Korean Journal of Food Science and Technology
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• v.45 no.3
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• pp.333-338
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• 2013

Enzymatic activity in germinated cereal grains is important for the saccharification of starch materials. This study was conducted to investigate the amylolytic activities of germinating brown rice and black rice that have different amylose contents. Brown rice and black rice were steeped at room temperature for 24 h and germinated at 20 and $30^{\circ}C$ for 1, 2, and 3 days. ${\alpha}$- and ${\beta}$-Amylase activities in normal brown rice increased very slightly during the 3-day germination period, but the enzymatic activities were slightly higher in low-amylose (waxy type) brown rice. Diastatic power (DP), a measure of starch-saccharifying enzyme, was higher in the germinating brown rice with low amylose than in those with normal amylose content. ${\alpha}$- and ${\beta}$-Amylase activities in black rice increased gradually during germination, and DP of low-amylose black rice appeared to be higher than that of normal brown rice. Amylase activities in brown rice and black rice germinated at $30^{\circ}C$ were higher than those germinated at $20^{\circ}C$. Compared to brown rice, the overall amylolytic activity of germinated black rice was observed to be higher than that of brown rice.

• Korean Journal of Food Science and Technology
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• v.33 no.3
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• pp.288-293
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• 2001

${\alpha}-amylase$ activities of bean water was not significantly influenced by bean water concentrations but they were remarkably influenced by different temperatures and substrates. ${\alpha}-amylase$ activities of bean water on cooked starch were significantly higher than those on raw starch. ${\beta}-amylase$ and glucoamylase activities in 14% bean water were significantly higher than those in 7% bean water. Yukwa paste is glutinous rice flour paste. Bean water was added to Yukwa paste by 0, 7, 14% and incubated 0, 3, 6, 9, 12 hours at $60^{\circ}C$. The pH of Yukwa paste increased with bean water concentration and decreased with the incubation time. The viscosity decreased with bean water concentration and incubation time. The ruducing sugar content of Yukwa paste increased with bean water concentration and incubation time. The changes of reducing sugar content in cooked Yukwa paste were much higher than those in the raw one. ${\alpha},\;{\beta}-amylase$glucoamylase activities of Yukwa paste also increased with bean water concentration, and their activities were much higher on the cooked glutinous rice flour than those on the raw one. The SEM observation on the freeze-dried flour of Yukwa paste showed breakdown of amylopectin structure by addition of bean water in the paste.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.1
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• pp.91-99
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• 2020

Objective: This study investigated the effects of oral administration of rumen-protected L-tryptophan (RPL-T) on duodenal starch digestion and gastrointestinal hormones (GIH) secretions using Hanwoo beef steers as the animal models. Methods: Four steers (423±24 kg) fitted with ruminal and duodenal cannulas were employed in a crossover design replicated twice. Treatments were control (basal diet) and RPL-T (basal diet+191.1 mg/kg body weight [BW]) group. Blood and duodenal samples were collected to measure serum GIH levels and pancreatic α-amylase activity at day 0, 1, 3, and 5 (-30, 30, 90, 150, and 210 min) of the study. Samples from each segment of the gastrointestinal tract were collected via ruminal and duodenal cannulas and were used to determine soluble protein and the starch digestion rate at days 6 (-30, 180, 360, and 540 min) and 8 (-30, 90, 270, and 450 min) of the experiment. Results: No significant difference in ruminal pH, NH₃-N, and total volatile fatty acid including the levels of acetate, propionate, butyrate, isobutyrate, valerate, isovalerate, and the acetate-to-propionate ratio was observed between groups (p>0.05). Crude protein uptake was higher and feces starch content was lower in RPL-T group than the control group (p<0.05). The D-glucose contents of feces in RPL-T group decreased at day 5 compared to those in the control group (p<0.05), however, no change was found at day 0, 1, or 3 compared to the control group (p>0.05). Serum cholecystokinin (CCK), melatonin, duodenal pancreatic α-amylase activity, and starch digestion were significantly higher in RPL-T group than the control group (p<0.05). Conclusion: Taken together, oral administration of RPL-T at the rate of 191.1 mg/kg BW consistently increased CCK concentration, pancreatic α-amylase activity in duodenal fluids, and starch digestion rate in the small intestine and thus found to be beneficial.

• Applied Biological Chemistry
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• v.48 no.1
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• pp.103-108
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• 2005

${\alpha}-Amylase$ inhibitor is used to control blood glucose level by inhibiting starch digestion in the small intestine and delaying the absorption of glucose. In this study, we investigated the effect of the ethanol extracts from more than 1400 species of plants against ${\alpha}-amylase$ with the aim of developing a new ${\alpha}-amylase$ inhibitor. In the results, Cornus walteri extracts showed the highest inhibition activity. The inhibitory effect of Cornus walteri extract on the carbohydrate hydrolysis enzymes has different sensitivities against ${\alpha}-amylase$ from salivary and pancreatin and against ${\alpha}-glucosidase$ from yeast and porcine small intestine. In the study of inhibition kinetics of ${\alpha}-amylase$ and ${\alpha}-glucosidase$, Cornus walteri extract showed competitive inhibition against salivary and pancreatin while showing the combination of uncompetitive and noncompetitive inhibition against ${\alpha}-glucosidase$. The Cornus walteri extract was stable at acidic and thermal conditions. As for the blood glucose and body weight levels of Cornus walteri extract, we confirmed anti-hyperglycemic and anti-obesity effects. Also, in the investigation of the mRNA lever, Cornus walteri extract upregulated the level of GLUT4 mRNA in the quadriceps muscle.

• Korean Journal of Pharmacognosy
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• v.31 no.1
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• pp.101-104
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• 2000

An inhibitory activity against ${\alpha}-glucosidase$ was identified in extract of an edible lichen, Umbilicaria esculenta. The inhibitor was very stable retaining above 95% of its original activity when treated with heat, acid and alkaline conditions, and some hydrolytic enzymes. Partially purified inhibitor showed strong inhibition against disaccharide hydrolytic enzymes of mammalian and mold origin, but weak or no inhibition against polysaccharide hydrolytic enzymes except glucoamylase. The inhibitor suppressed elevation of blood glucose level in rats after oral administration of soluble starch or sucrose.

• BMB Reports
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• v.37 no.6
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• pp.642-647
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• 2004

The lysine residues of Bacillus licheniformis $\alpha$-amylase (BLA) were chemically modified using citraconic anhydride or succinic anhydride. Modification caused fundamental changes in the enzymes specificity, as indicated by a dramatic increase in maltosidase and a reduction in amylase activity. These changes in substrate specificity were found to coincide with a change in the cleavage pattern of the substrates and with a conversion of the native endo- form of the enzyme to a modified exo- form. Progressive increases in the productions of $\rho$-nitrophenol or glucose, when para nitrophenyl-maltoheptaoside or soluble starch, respectively, was used as substrate, were observed upon modification. The described changes were affected by the size of incorporated modified reagent: citraconic anhydride was more effective than succinic anhydride. Reasons for the observed changes are discussed and reasons for the effectivenesses of chemical modifications for tailoring enzyme specificities are suggested.