• Microbiology and Biotechnology Letters
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• v.17 no.1
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• pp.14-18
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• 1989

As the final experiment to assess the possibility of industrial application of FSC14-75, ethanol productivity from liquefied sweet potato starch was examined in a pilot scale of 300 liters. FSC14-75 produced 6.6％(v/v) of ethanol from 13.3％ of liquefied sweet potato starch in 8 days, and the residual sugar was 3.15％. The corresponding efficiency was 70％ of the theoretical maximum. Since we could isolate unicellular cell and flocculent cell from the fermentation broth, we designated them FSC14-75(S) and FSC14-75(F), respectively. We investigated ethanol productivity of FSC14-75(F) compared with that of FSC14-75(S) from liquefied potato starch in a mini·tar tormentor scale of 2.5 liters. FSC14-75(F) was found more favorable than the counterpart in terms of ethanol productivity, and produced 8.1％(v/v) of ethanol from 15％ of liquefied potato starch with an efficiency of 75％. In a pilot scale fermentation with 15％ of liquefied sweet potato starch, ethanol productivity of FSC14-75(F) reached maximum level of 7.7％(v/v) after 8 days, and the residual sugar was 1.9％. However, the ethanol productivity was not enhanced by a supplementary addition of Thermamyl to the fermentation broth after sterilization.

• The Korean Journal of Mycology
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• v.24 no.1 s.76
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• pp.8-16
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• 1996

The ${\alpha}-amylase$ of Alternaria alternata was purified through ammonium sulfate precipitation, dialysis and Sephadex G-100 column chromatography. One single band was obtained in SDS-polyacrylamide gel electrophoresis. The optimum pH for enzyme activity was 5.0 and the enzyme activity was maintained at $3.6{\sim}7.0$pH range. The optimum temperature for ${\alpha}-amylase$ activity was $40^{\circ}C$ and 71% of the activity was still maintained until 30 min after heating at $80^{\circ}C$. The ${\alpha}-amylase$ was slightly activated by $Mn^{2+},\;Zn^{2+}\;and\;Sn^{2+}$, but inhibited by $Ba^{2+},\;Pb^{2+},\;Co^{2+}\;and\;Ag^{1+}$. The $Hg^{2+}\;and\;Ag^{2+}$ slightly inhibited the activity of the enzyme at concentrations of $10^{-3}\;and\;10^{-4}M$. The Michaelis constant $(K_m)$ to soluble starch was $6.50{\times}10^{-2}M$ and inhibition constant $(K_i)$ by the 1mM EDTA was $8.0{\times}10^{-2}M$. The inhibition of this enzyme by EDTA was competitive one.

• Journal of Microbiology and Biotechnology
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• v.22 no.8
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• pp.1077-1083
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• 2012

Previously, an extracellular ${\alpha}$-amylase (BKA) had been purified from the culture of Bacillus sp. KR8104. Subsequently, the crystal structure of the active enzyme revealed a 422 amino acids polypeptide. In this study, the bka was cloned into E. coli, which encoded a polypeptide of 659 amino acids including two additional fragments: one 44 residues N-terminal fragment and another 193 residues C-terminal fragment. In order to investigate the role of the C-terminal fragment, two constructs with and without this region [$BKA{\Delta}$(N44) and $BKA{\Delta}$(N44C193)] were designed and expressed in E. coli BL21. The optimum pH, thermal stability, and the end-products of starch hydrolysis were found to be similar in both constructs. The $K_m$ and $V_{max}$ values for $BKA{\Delta}$(N44) were lower than $BKA{\Delta}$(N44C193), using either starch or ethylidene-blocked 4-nitrophenylmaltoheptaoside as a substrate.

• Korean Journal of Food Science and Technology
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• v.37 no.5
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• pp.763-767
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• 2005

Effect of ${\alpha}-amylase$ and various emulsifiers on characteristics of bread dough were examined. Fungal or bacterial ${\alpha}-amylase$ and various emulsifiers including monoglyceride (MG), sodium stearoyl-2-lactylate (SSL), and diacetyltartaric acid ester of mono- and diglycerides (DATEM) were added to bread dough both individually and as mixtures. Rheological characteristics of various bread doughs were examined through falling number, farinograph, alveograph, and rapid visco analysis. Results obtained showed falling number decreased via degradation of starch by ${\alpha}-amylase$. In farinogram, addition of ${\alpha}-amylase$ and emulsifiers in dough decreased consistency, water absorption, mechanical tolerance index, and dough development time. Farinogram characteristic was improved by adding SSL+MG to dough formula. Similar to farinogram addition of ${\alpha}-amylase$ and emulsifiers in alveogram of dough decreased overpressure, extensibility, swelling index, and deformation energy. Whereas addition of ${\alpha}-amylase$ did not affect pasting temperature, viscosity of dough tended to decrease.

• Preventive Nutrition and Food Science
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• v.18 no.3
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• pp.163-168
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• 2013

In this study, we investigated the inhibitory effects of Baechu kimchi added Ecklonia cava on the activities of ${\alpha}$-glucosidase and ${\alpha}$-amylase and its alleviating effect on the postprandial hyperglycemia in STZ-induced diabetic mice. Baechu kimchi added Ecklonia cava (BKE, 15%) was fermented at $5^{\circ}C$ for 28 days. Optimum ripened BKE was used in this study as it showedthe strongest inhibitory activities on ${\alpha}$-glucosidase and ${\alpha}$-amylaseby fermentation time among the BKEs in our previous study. The BKE was extracted with 80% methanol and the extract solution was concentrated, and then used in this study. The BKE extract showed higher inhibitory activities than Baechu kimchi extract against ${\alpha}$-glucosidase and ${\alpha}$-amylase. The $IC_{50}$ values of the BKE extract against ${\alpha}$-glucosidase and ${\alpha}$-amylase were 0.58 and 0.35 mg/mL, respectively; BKE exhibited a lower ${\alpha}$-glucosidase inhibitory activity but a higher ${\alpha}$-amylase inhibitory activity than those of acarbose. The BKE extract alleviated postprandial hyperglycemia caused by starch loading in normal and streptozotocin- induced diabetic mice. Furthermore, the BKE extract significantly lowered the incremental area under the curve in both normal and diabetic mice (P<0.05). These results indicated that the BKE extract may delay carbohydrate digestion and thus glucose absorption.

• Journal of the Korean Society of Food Science and Nutrition
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• v.16 no.2
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• pp.128-135
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• 1987

Extracellular ${\alpha}-amylase$, ${\beta}-amylase$ and glucoamylase produced by a thermophilic and cellulolytic bacterium, Herpetosiphon geysericola CUM 317, were partially purified by salting out with ammonium sulfate and by chromatography on a DEAE-cellulose column and on a CM-cellulose column. The Km values of ${\alpha}-amylase$, ${\beta}-amylase$ and glucoamylase for potato starch were $2.31mg/m{\ell}$, $7.69mg/m{\ell}$, and $8.33mg/m{\ell}$. The molecular weights of ${\alpha}-amylase$, ${\beta}-amylase$ and glucoamylase were calculated to be about 84000 dalton, 76000 dalton and 80000 dalton, respectively.

• Microbiology and Biotechnology Letters
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• v.20 no.2
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• pp.143-149
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• 1992

A bacterial strain No. 2, which highly produced a-glucosidase, was isolated from Kimchi and identified to be a similar species of Pediococcus halophilus. This enzyme was purified by protamine sulfate, ammonium sulfate fractionation, ion exchange and gel filtration. The maximal a-glucosidase activity was observed at pH 6.0 and this enzyme was stable at pH 6.0~ 7.5. The optimum temperature of this enzyme activity was $37^{\circ}C$, but enzyme activity was gradually lost above $37^{\circ}C$. This enzyme was activated by 10 mM MgCh and inhibited by 10 mM mercaptoethanol. The kinetics of PNPG(p-Nitrophenyl-a-D-glucopyranoside) and maltose were Kp0.52 mM/27.5 pg protein, $V_{max}$= 0.021 mM/min 27.5 ${\mu}g$ protein and $K_m$= 0.32 mMD7.5 ${\mu}g$ protein, $V_{max}$= 0.025 mM/min 27.5 ${\mu}g$ protein, respectively. The molecular weight of $\alpha$-glucosidase was about 37, 000.

• Korean Journal of Food Science and Technology
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• v.24 no.1
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• pp.37-41
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• 1992

Starch solid system was used to investigate the effects of lipoxygenase, linoleic acid and water activity on the oxidation of ${\beta}-carotene$ or ${\alpha}-tocopherol$. ${\beta}-carotene$ or ${\alpha}-tocopherol$ was co-oxidized severely with linoleic acid by lipoxygenase, and these were reduced to 19% and 5% of initial concentration, respectively, after 2 days storage at $a_w$ 0.72 in the system. The concentration of ${\beta}-carotene$ and the destruction rates were linearly correlated. However, the ${\beta}-carotene$ was very stable in the system without linoleic acid and lipoxygenase. The oxidation products of ${\alpha}-tocopherol$ were considered as ${\alpha}-tocopheryl$ quinone and ${\alpha}-tocopheryl$ dimer, and the level of ${\alpha}-tocopherol$ quinone increased as the reaction time increased.

• Korean Journal of Food Science and Technology
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• v.42 no.2
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• pp.198-203
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• 2010

Although barley $\alpha$-amylase isozyme 1 (AMY1) and 2 (AMY2) share up to 80% of amino acid sequence identity, their enzymatic properties differ remarkably. In this study, the 42nd alanine residue of AMY2 was replaced with another random amino acid via saturation mutagenesis. Eight out of 370 recombinant E. coli cells showing enhanced starch-hydrolyzing activity were characterized as possessing the same proline residue instead of alanine. Even though the specific activity of AMY2-A42P is reduced to 81% of wild-type, its expression level and purification yield were enhanced by approximately 2 and 4 times that of AMY2, respectively. Characterization of its enzymatic properties confirmed that AMY2-A42P is similar to that of wild-type. However, its specificity to starch substrates is likely to be intermediate between AMY1 and AMY2.

• BMB Reports
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• v.28 no.5
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• pp.375-381
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• 1995

Two forms of glucoamylase (GI and GII) from starch-grown Lipomyces kononenkoae CBS 5608 mutant were purified to apparent homogeneity by means of ultrafiltration, Sephacryl S-200 gel filtration and DEAE Sephadex A-50 chromatography. The apparent molecular weight was calculated as ca. 150 kDa for GI and ca. 128 kDa for GII, respectively. Both enzymes were glycoproteins with isoelectric points of 5.6 (GI) and 5.4 (GII). They had a pH optimun of 4.5 and were stable from pH 5 to 8. The temperature optimum for both enzymes was $60^{\circ}C$, but they were rapidly inactivated above $70^{\circ}C$. The $K_m$ values toward starch were estimated to be 6.57 mg per ml for GI and 4.52 mg per ml for GII, and the $V_{max}$ values were 16.28 ${\mu}M$ per mg for GI and 32.25 ${\mu}M$ per mg for GII, respectively. The $K_m$ and $V_{max}$ values of GII for ${\alpha}-$ or ${\beta}-cyclodextrin$ were estimated to be 0.15 mg per ml and 2.0 mg per ml, respectively ($K_m$) and 1.02 ${\mu}M$ per mg or 1.02 ${\mu}M$ per mg, respectively ($V_{max}$). Neither enzyme exhibited pullulanase activity but they released only glucose from starch or cyclodextrin. Amino acid analysis indicated that both glucoamylases were enriched in proline and acid amino acids. Glucoamylase GII strongly cross-reacted with a monoclonal antibody raised against GI enzymes, and the two enzymes shared very similar amino acid composition. Western blot analysis indicated that L. kononenkoae CBS 5608 mutant produced two forms of glucoamylase on starch, and that synthesis of them was subject to glucose repression.