• Preventive Nutrition and Food Science
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• v.13 no.3
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• pp.190-195
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• 2008

The $\alpha$-amylase gene, amyL, from Bacillus licheniformis was expressed in Lactobacillus brevis 2.14 and Escherichia coli $DH5{\alpha}$ using two different shuttle vectors, pCW4 and pSJE. E. coli transformants (TFs) harboring either $pCW4T{\alpha}$ or $pSJET{\alpha}$ produced active $\alpha$-amylase but L. brevis TFs did not, as determined by enzyme assays and zymography. But amyL transcripts were synthesized in L. brevis TFs. In terms of plasmid stability, pSJE, a theta-type replicon, was more stable than pCW4, an RCR (rolling circle replication) plasmid, in L. brevis without antibiotic selection.

• Journal of Applied Biological Chemistry
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• v.50 no.2
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• pp.70-73
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• 2007

Antioxidant and anti-diabetic activities of acorn were evaluated by its potential for scavenging stable DPPH free radical, inhibition of lipid peroxidation, reducing power, and inhibiton of ${\alpha}-glucosidase\;and\;{\alpha}-amylase$. The water extract of acorn exhibited strong antioxidant and antidiabetic related activities in the tested model systems. Solvent fractionation of the water extract revealed that the water fraction and the EtOAc fraction had strong antioxidant activity, and inhibitory activity on ${\alpha}-glucosidase\;and\;{\alpha}-amylase$. The water fraction exhibited higher DPPH radical scavenging activity ($EC_{50}=7.19{\mu}g/mL$) than that of ${\alpha}-tocopherol\;(EC_{50}=7.59{\mu}g/mL)$. It is considered that water extract of acorn has the potential for natural antioxidant and anti-diabetic products.

• Journal of Microbiology and Biotechnology
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• v.24 no.2
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• pp.254-263
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• 2014

Enteromorpha polysaccharides (EP) extracted from green algae have displayed a wide variety of biological activities. However, their high molecular weight leads to a high viscosity and low solubility, and therefore, greatly restrains their application. To solve this problem, bacteria from the surface of Enteromorpha were screened, and an Alteromonas macleodii strain B7 was found to be able to decrease the molecular weight of EP in culture media. Proteins harvested from the supernatant of the A. macleodii B7 culture were subjected to native gel electrophoresis, and a band corresponding to the Enteromorpha polysaccharide lyase (EPL) was detected by activity staining. The enzyme identity was subsequently confirmed by MALDI-TOF/TOF mass spectrometry as the putative ${\alpha}$-amylase reported in A. macleodii ATCC 27126. The amylase gene (amySTU) from A. macleodii B7 was cloned into Escherichia coli, resulting in high-level expression of the recombinant enzyme with EP-degrading activity. AmySTU was found to be cold-adapted; however, its optimal enzyme activity was detected at $40^{\circ}C$. The ${\alpha}$-amylase was highly stable over a broad pH range (5.5-10) with the optimal pH at 7.5-8.0. The highest enzyme activity was detected when NaCl concentration was 2%, which dropped by 50% when the NaCl concentration was increased to 16%, showing an excellent nature of halotolerance. Furthermore, the amylase activity was not significantly affected by tested surfactants or the presence of some organic solvents. Therefore, the A. macleodii strain B7 and its ${\alpha}$-amylase can be useful in lowering EP molecular weight and in starch processing.

• Journal of Life Science
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• v.7 no.1
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• pp.30-38
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• 1997

For screening thermostable $\alpha$-amylase from thermophiles, various samples from extreme environments such as hot spring and sewage near them, and compoat, wereexamined microbial growth in enrichment culture medium at 55$\circ$C on the assumption that enzymes from thermophiles are inevitable thermostable. One strain showing higher $\alpha$-amylase activity was pure cultured and designated as Bacillus sp. TR-25 from the results of morphological, cultural and physiological characteristics. The most important carbon sourses for the enzyme production were soluble starch, dextrin, potato starch and corn starch. Glucose and fructose had a catabolite repression on the enzyme production. The good nitrogen sources for the enzyme production were yeat extract, nutrient broth, tryptone, corn steep liquor and ammonium sulfate. The enzyme production was accelerated by addition of CaCl$_{2}$. $\cdot $ H$_{2}$O. The optimal medium composition for the enzyme production was soluble starch 2.0%, yeast extract 0.55, CaCl$_{2}$ $\cdot $ 2H$_{2}$O 0.015, Tween 80 0.001%, pH8.0, respectively. In jar fermenter culture, this strain shows a rapid growth and required cheaper carbon and nitrogen source. These properties are very useful to fermentation industry. The $\alpha$-amylase of this strain demonstrated a maximum activity at 80$\circ$C, pH 5.0, respectively. And calcium ion did not improve thermostability of the enzyme. At 10$0^{\circ}C$, this enzyme has 235 of relative activity. Transformation was carried out by thermophilic Bacillus sp. TR-25 genomic DNA. As a result, the transformant has increased thermostable $\alpha$-amylase activity.

• Journal of Microbiology and Biotechnology
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• v.23 no.2
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• pp.205-210
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• 2013

Thiol-ene polymerization is a versatile tool for several applications. Here we report the preparation of epoxide groups containing thiol-ene photocurable polymeric support and the covalent immobilization of ${\alpha}$-amylase onto these polymeric materials. The morphology of the polymeric support was characterized by scanning electron microscopy (SEM), and energy dispersive spectroscopy (EDS) coupled with SEM was used to explore the chemical composition. The polymeric support and the immobilization of the enzyme were characterized by FTIR analysis. SEM-EDS and FTIR results showed that the enzyme was successfully covalently attached to the polymeric support. The immobilization efficiency and enzyme activity of ${\alpha}$-amylase were examined at various pH (5.0-8.0) and temperature ($30-80^{\circ}C$) values. The storage stability and reusability of immobilized ${\alpha}$-amylase were investigated. The immobilization yield was $276{\pm}1.6$ mg per gram of polymeric support. Enzyme assays demonstrated that the immobilized enzyme exhibited better thermostability than the free one. The storage stability and reusability were improved by the immobilization on this enzyme support. Free enzyme lost its activity completely within 15 days. On the other hand, the immobilized enzyme retained 86.7% of its activity after 30 days. These results confirm that ${\alpha}$-amylase was successfully immobilized and gained a more stable character compared with the free one.

• Journal of Sericultural and Entomological Science
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• v.21 no.1
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• pp.30-35
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• 1979

This experiment was conducted to investigate the effects of $\alpha$-amylase, added to artificial diet, on the larval growth and on the amount of blood sugar of the silkworm Bombyx mori L.. The artificial diet containing 20% mulberry leaf powder was fed on the 5th instar silkworm. The results obtained were summarized follows: 1. The digestibility in both sexes was increased by feeding artificial diet with $\alpha$-amylase and increase of the digestibility was greater in the female than in the male. 2. Larval body weight was increased in the female but was the male, compared with those in each sex of the control, respectively. 3. There was an increase of the single cocoon weight and cocoon shell weight, as compared with that of the control. 4. It was evident that the artificial diet containing $\alpha$-amylase was more effective than the control, and the effect of the diet was considerable higher in the female than in the male. 5. The amount of the blood sugar on the 7th day of 5th instar was decreased as compared with that of the control.

• Microbiology and Biotechnology Letters
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• v.47 no.3
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• pp.412-422
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• 2019

The present study reports the morphological and molecular characterization of the fungal strain, CMSS06 and evaluates its raw starch hydrolyzing ability in four different agricultural substrates (rice bran, banana peel, cassava tubers, and coconut water). The potential use of each agricultural substrate to replace the expensive fermentation media was evaluated with six different fermentation media: rice bran (RB), banana peel (BP), cassava starch (CS), cassava in coconut water (CSCW), cassava in modified coconut water (CMCW), and pure Coconut water (CW). The fungal strain CMSS06 was identified as Thielaviopsis ethacetica by the analysis of the ITS sequences. The T. ethacetica alpha amylase enzyme exhibited maximum alpha amylase activity at 72 h, pH 7.0, and $40^{\circ}C$ on soluble starch. This species resulted in the highest enzyme activity (mU/ml) of 26.06, 10.89, 58.82, 14.2, and 54.67 with the RB, BP, CS, CSCW, and CMCW fermentation media, respectively. The results indicate that CS can be used as a carbon substrate and CMCW can be used to accelerate the fermentation by T. ethacetica. The enzyme was partially purified by 40-60% ammonium sulphate fraction, and it showed total enzyme activity, total protein content, specific activity, purification fold, and a recovery of 2400 mU, 30 mg, 80 mU/mg, 2.7, and 71.1%, respectively. The molecular mass of the T. ethacetica alpha amylase was estimated on SDS-PAGE, and two bands around 50 kDa and 70 kDa were identified. The present study implies that T. ethacetica can produce alpha amylase, and it can be used to hydrolyze raw starch during the fermentation processes.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.50 no.6
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• pp.378-383
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• 2005

This study was conducted in order to exam­ine relationships between the viviparous germination ability and the antioxidative system in rice seeds and seedlings. Enzyme activities was compared among the four varieties with different viviparous germination ability from the early or mid-late maturity group. $\alpha$-Amylase activities correlated with the viviparous germination rate (VGRs) and $\alpha$-amylase activity in seeds of 40 days after heading (DAH) was highest in Dongjinbyeo among mid­late maturing varieties (MLMVs) and in Daeseongbyeo among EMVs. $\alpha$-Amylase activity in dry mature seeds was also higher in varieties with higher VGR. Glucose contents in viviparously germinating seeds and mature dry, seeds were higher in varieties with higher VGRs. Cat-alase activities did not correlate with the VGRs in both maturity groups. However, peroxidase activities in viviparously germinating seeds were higher in varieties with higher VGRs.

• Microbiology and Biotechnology Letters
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• v.16 no.5
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• pp.348-351
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• 1988

The production of extracellular $\alpha$-amylase by Bacillus thuringiensis 19 serotypes was examined by the hydrolysis test of starch. Thirteen serotypes produced the amylase. B. thuringiensis serovar thuringiensis alesti, kurstaki, sotto, kenya, israelensis, morrisoni, entomocidus, tolworthi, thompsoni, toumanoffi, pakistani, and indiana produced tee enzyme. The amylase produced by B. thuringiensis serovar israelensis showed highest activity around pH 6.7 to 7.2 and 55$^{\circ}C$ to $65^{\circ}C$. The high production medium of the amylase was composed of 1% bactopeptone, 0.3% beef-extract, 0.3% yeast ex-tract, 0.5% NaCl, 0.3% $K_2$HPO$_4$, 0.1% KH$_2$PO$_4$, 0.2% Soluble Starch, 0.012% CaCl$_2$.2$H_2O$$_2$, 0.005% MnCl$_2$, and 0.03% MgCl$_2$.7$H_2O$. The highest production of the enzyme was observed at 4 hours culture in the soluble starch (0.6 units/$m\ell$) or glucose (0.43 units/$m\ell$) substrate.

• Preventive Nutrition and Food Science
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• v.7 no.1
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• pp.62-66
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• 2002

As inhibitors of carbohydrate-digesting enzymes can prevent hyperglycemia that is known to cause many macrovascular complications, they may prove a useful adjunct to hypocaloric diets in patients with type 2 diabetes and obesity. Inhibitory activities of two hundred and fifteen kinds of medicinal herb extracts against $\alpha$-glucosidase (EC 3.2.1.20) and $\alpha$-amylase (EC 3.2.1.1) have been investigated in vitro. Adenophora triphylla, Aneilema keisak, and Morus bombysis significantly suppressed rat intestinal $\alpha$-glucosidase activity iu vitro. Porcine pancreatic amylase was efficiently inhibited by methanol extracts of Epimedium koreanum, Campsis grandiflora and Salvia plebeia. Methanol extract of Epimedium koreanum among the medicinal herbs tested showed the strongest inhibitory activity against porcine pancreatic $\alpha$-amylase with 0.1 mg/ML of $IC_{50}$/. The herb extract also improved glucose tolerance in ICR mice when loaded with 0.9 g soluble starch per kg body weight. Taken together, Epimedium koreanum merits further evaluation as a therapeutic measure.