• Applied Biological Chemistry
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• v.21 no.2
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• pp.103-108
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• 1978

Acid-stable ${\alpha}-amylase$ was partially purified from Paecilomyces subglobosum by Sephadex G-150 gel filtration. About 7.7-fold purification was obtained and the partially purified preparation has 5.0 U of ${\alpha}-amylase$ activity per mg of protein. Using this partially purified ${\alpha}-amylase$, general properties were studied and it showed the maximal activities at the conditions of pH 4.0 and $38^{\circ}C$. High stability of the acid-stable ${\alpha}-amylase$ in acidic condition was observed, whereas thermal stability was similar to the conventional ${\alpha}-amylase$.

• Bulletin of the Korean Chemical Society
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• v.14 no.3
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• pp.398-403
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• 1993

• Journal of Life Science
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• v.25 no.8
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• pp.870-879
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• 2015

The objective of this research was to evaluate the inhibitory activities of phenolic compounds isolated from mulberry (Morus alba) leaves of 109 types against α-amylase and α-glucosidase. The inhibitory activity of the water extracts from Morus alba cv. Kuksang against α-amylase and α-glucosidase were determined as 93.8% and 48.7% respectively. The total phenolic content of extracts from Morus alba cv. Kuksang was 9.7±0.2 mg/g soluble in water and 14.3±0.2 mg/g soluble in ethanol. The inhibitory activity of the water extracts from Morus alba cv. Kuksang at 200 μg/ml phenolics concentration against α-amylase and α-glucosidase were determined as 100% and 82.6% respectively. The purification of inhibitory compounds was carried out by Sephadex LH-20 and MCI-gel CHP-20 column chromatography using a gradient elution procedure by nomal phase type (EtOH→distilled water) and reverse phase type (distilled water→MeOH). The quercetin was confirmed to be the chemical structure of the inhibitory compound against α-amylase and α-glucosidase by spectroscopic analysis of FAB-MS, NMR and IR spectrum.

• The Journal of the Korean dental association
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• v.10 no.12
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• pp.803-807
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• 1972

사람타액의 Amylase 부분 정제를 위하여 각 %의 Ammonium Sulfate 포화획분을 얻은 다음 Calcium phosphate gel로 처리하고 각 농도의 Na₂HPO₄로 유출시킨 일차실험결과는 다음과 같다. 1. Ammonium Sulfate 포화획분중 50-70% 포화획분의 Amylase활성이 가장 높았다. 2. Amylase 비활성은 0.02M Na₂HPO₄로 유출한 획분에서 가장 높았다.

• Microbiology and Biotechnology Letters
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• v.9 no.2
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• pp.91-97
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• 1981

A thermophilic soil isolate Bacillus sp. Y-127 was selected for the production of thermostable amylase. The strain was used for the enzyme production and the thermostable amylase was characterized. The optimum cultural conditions for the enzyme production were 6$0^{\circ}C$ at pH 7.0 for 32 hours using a mineral medium containing 2% soluble starch and 0.2% yeast extract. The extra-cellular enzyme was purified about 123-folds with about 6% recovery. The purified enzyme was stable at pH between 4.0 and 7.0, and temperature up to 6$0^{\circ}C$.

• Journal of Microbiology and Biotechnology
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• v.21 no.5
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• pp.470-476
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• 2011

A potent fungus for amylase production, Chrysosporium asperatum, was isolated from among 30 different cultures obtained from wood samples collected in the Junagadh forest, India. All of the isolated cultures were screened for their ability to produce amylase by submerged fermentation. Among the selected cultures, C. asperatum (Class Euascomycetes; Onygenales; Onygenaceae) gave maximum amylase production. In all of the different media tested, potato starch was found to be a good substrate for production of amylase enzyme at $30^{\circ}C$ and pH 5.0. Production of enzyme reached the maximum when a combination of starch and 2% xylose, and organic nitrogen (1% yeast extract) and ammonium sulfate were used as carbon and nitrogen sources, respectively. There was no significant effect of metal ions on enzyme activity. The enzyme was relatively stable at $30^{\circ}C$ for 20 min, and no inhibitory effect of $Ca^{+2}$ ions on amylase production was observed.

• Bulletin of the Korean Chemical Society
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• v.15 no.10
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• pp.832-835
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• 1994

The ${alpha}$-amylase gene of Bacillus licheniformis has been cloned and two mutant ${alpha}$-amylase genes of which histidine 235 was changed to glutamine (H235Q) and aspartic acid 328 to glutamic acid (D328E) have been produced by site-directed mutagenesis. The kinetic parameters, optimum pH and thermostability of wild type(WT) and these two mutant amylases expressed in E. coli MC1061 have been compared after purification. The $K_m$ values of WT, H235Q and D328E ${alpha}$-amylases were 0.22%, 0.73%, and 0.80% respectively, when using starch as the substrate. The $V_max$ values of wild type ${alpha}$ -amylase and mutant ${alpha}$-amylases were 0.6-0.7%/minute, and did not show any significant differences among them. The optimum pH of D328E ${alpha}$-amylase was shifted to more acidic pH. Also, the thermostability of H235Q ${alpha}$-amylase was increased compared to the wild type ${alpha}$-amylase.

• Microbiology and Biotechnology Letters
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• v.27 no.2
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• pp.129-135
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• 1999

EH-1 was highest at 9 days of incubation. This regrowth and enzymatic activity of Haloarcular sp. EH-1 was highest at 9 days of incubation. This amylase was purified by acetone fractionation, DEAG-Cellulose column chromatography, 1st Sephadex G-75 gel filtration, CM-Cellulose column chromatography and 2nd Sephadex G-75 gel filtration. The amylase was purified about 98.64 fold with a yield of 11.75%. The molecular weight of amylase was estimated to be about 43,000and 40,000 by gel filtration and SDS-polyacrylamide gel electrophoresis, respectively, suggesting that the enzyme was a monomer. Amylase had an optimal temperature of 4$0^{\circ}C$, and an optimum pH of 7.0, and the thermal stability was observed the above 50% at 10$0^{\circ}C$ after 1 hour, and the stable range of pH was 6.0 to 8.0. The enzymatic activity was increased in the presence of 10 mM 2-mercaptoethanol, slightly by 10 mM SnCl2.2H2O.FeCl2.4H2O.CuCl2.2H2O.HgCl2.6H2O and SDS. End products from soluble starch were glucose, maltose and maltotriose, and Km value for soluble starch was 2.5mg/ml.

• Korean Journal of Food Science and Technology
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• v.33 no.1
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• pp.117-121
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• 2001

White kidney bean, Phaseolus vulgaris, contains proteinaceous inhibitors of ${\alpha}-amylase$. Two inhibitors have been purified by conventional protein fractionation methods such as ethanol precipitation, ammonium sulfate fractionation, DEAE-Sephadex ion exchange chromatography and Sephadex G-100 gel chromatography. The inhibitors were purified as I-1 and I-2 based on their elution order from the DEAE-Sephadex column. The overall purification ratio were about 15.0 and 14.8 for I-1 and I-2, respectively. The molecular weights of purified ${\alpha}-amylase$ inhibitors were 50,000 and 45,000 determined by Sephadex G-100 gel filtration and SDS-polyacrylamide gel electrophoresis. They contain $17.6{\sim}17%$ of carbohydrate, $70{\sim}80%$ of protein. The carbohydrates were composed of glucose : xylose : mannose : N-acetylglucosamine (5 : 3 : 50 : 42).

• BMB Reports
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• v.40 no.4
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• pp.494-500
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• 2007

The endophytic bruchid pest Callosobruchus maculatus causes severe damage to storage cowpea seeds, leading to economical losses. For this reason the use of $\alpha$-amylase inhibitors to interfere with the pest digestion process has been an interesting alternative to control bruchids. With this aim, $\alpha$-amylase inhibitors from baru seeds (Dipteryx alata) were isolated by affinity chromatographic procedures, causing enhanced inhibition of C. maculatus and Anthonomus grandis $\alpha$-amylases. To attempt further purification, this fraction was applied onto a reversed-phase HPLC column, generating four peaks with remarkable inhibition toward C. maculatus $\alpha$-amylases. SDS-PAGE and MALDI-ToF analysis identified major proteins of approximately 5.0, 11.0, 20.0 and 55 kDa that showed $\alpha$-amylase inhibition. Results of in vivo bioassays using artificial seeds containing 1.0% (w/w) of baru crude extract revealed 40% cowpea weevil larvae mortality. These results provide evidence that several $\alpha$-amylase inhibitors classes, with biotechnological potential, can be isolated from a single plant species.