• BMB Reports
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• v.40 no.3
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• pp.315-324
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• 2007

The novel $\alpha$-amylase purified from locally isolated strain, Bacillus sp. KR-8104, (KRA) (Enzyme Microb Technol; 2005; 36: 666-671) is active in a wide range of pH. The enzyme maximum activity is at pH 4.0 and it retains 90% of activity at pH 3.5. The irreversible thermoinactivation patterns of KRA and the enzyme activity are not changed in the presence and absence of $Ca^{2+}$ and EDTA. Therefore, KRA acts as a $Ca^{2+}$-independent enzyme. Based on circular dichroism (CD) data from thermal unfolding of the enzyme recorded at 222 nm, addition of $Ca^{2+}$ and EDTA similar to its irreversible thermoinactivation, does not influence the thermal denaturation of the enzyme and its Tm. The amino acid sequence of KRA was obtained from the nucleotide sequencing of PCR products of encoding gene. The deduced amino acid sequence of the enzyme revealed a very high sequence homology to Bacillus amyloliquefaciens (BAA) (85% identity, 90% similarity) and Bacillus licheniformis $\alpha$-amylases (BLA) (81% identity, 88% similarity). To elucidate and understand these characteristics of the $\alpha$-amylase, a model of 3D structure of KRA was constructed using the crystal structure of the mutant of BLA as the platform and refined with a molecular dynamics (MD) simulation program. Interestingly enough, there is only one amino acid substitution for KRA in comparison with BLA and BAA in the region involved in the calcium-binding sites. On the other hand, there are many amino acid differences between BLA and KRA at the interface of A and B domains and around the metal triad and active site area. These alterations could have a role in stabilizing the native structure of the loop in the active site cleft and maintenance and stabilization of the putative metal triad-binding site. The amino acid differences at the active site cleft and around the catalytic residues might affect their pKa values and consequently shift its pH profile. In addition, the intrinsic fluorescence intensity of the enzyme at 350 nm does not show considerable change at pH 3.5-7.0.

• The Korean Journal of Food And Nutrition
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• v.30 no.3
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• pp.491-500
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• 2017

Sweet potato-malt worts were prepared by using sweet potato paste of Shinyulmi and Shinjami as the main adjunct, enzymes, malt, hop, and water. We brewed low-malt beers of the lager- or ale-type by using these worts and inoculating bottom and top fermenting yeast, respectively. Moreover, the componential and functional characteristics of the resulting beers were evaluated. During saccharification of sweet potato, the addition of an enzyme agent containing ${\alpha}-amylase$ caused an improvement in filterability and an increase of total sugar. The sugar content of sweet potato-malt wort which was prepared by the addition of 0.1% enzyme agent containing ${\alpha}-amylase$ and a three-step infusion procedure was $13.5^{\circ}Brix$ adequate for beer brewing. The polyphenol and anthocyanin contents of Shinjami beer were increased with increasing content of the paste, and also increased DPPH and ABTS radical scavenging activities. But in Shinyulmi beer it were decreased. A strong correlation was obserbed between antioxidave activities and polyphenol and anthocyanin contents of sweet potato beers. In all lager- and ale-type low-malt beers using 41.6% of Shinjami pastes, sensory attributes very similar to those of 100% malt beer were obtained and they were very good as they had unique red color.

• Korean Journal of Soil Science and Fertilizer
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• v.26 no.3
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• pp.189-196
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• 1993

Allelopathic inhibition by substance(s) originated from sicklepod(Cassia tora L.) seed on germination of other plant seeds, was confirmed and used to elucidate general mechanism of allelopathy which was occurred widly in natural and/or agricultural ecosystems. The mechanism was deduced from changes in water contents, ${\alpha}$-amylase activities, protease activities, concentrations of abscisic acid and total phenolic compounds during germination of rice and radish seeds treated with water-soluble extracts of sicklepod seeds. The results obtained were summarized as follows. 1. The % germinations of radish, rice, lettuce, barley seeds were decreased by substances originated from crushed sicklepod seeds. 2. By the treatment of water-soluble extracts of sicklepod seeds, the inhibition of germination of radish seeds was occured with the sequential phenomena of increase of protease activity(synthesis), decrease of water content and increase of total phenolic compounds content. 3. In rice, the inhibition of seed germination by the water-slouble extracts of sicklepod seeds was related to increase of abscisic acid concentration and then decrease of ${\alpha}$-amylase activity(synthesis).

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.11
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• pp.1479-1484
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• 2008

This study was conducted to investigate the quality characteristics of fermented-defatted soybean grits (DSG). Fermented-DSG was prepared by inoculating Bacillus subtilis NUCI into steamed DSG at 2% (v/w) concentration and fermenting at $40^{\circ}C$ for 24 hr. The protein contents of DSG and fermented-DSG were higher by $57.2{\sim}61.0%$ than those of Chungkukjang, but lipid contents of these samples were not detected. The protease and $\alpha$-amylase activities, and solid contents in DSG were not shown. However, the activities of protease and $\alpha$-amylase, tyrosine contents, fluid consistency, and viscous substance contents of fermented-DSG were higher than those of Chungkukjang. The levels of free amino acids and isoflavone of fermented-DSG tended to increase more than those of DSG. The sensory score of fermented-DSG were higher in odor and taste than those of Chungkukjang groups, while lower in color and overall acceptability.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.6
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• pp.761-766
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• 2008

The effects of various enzymes and emulsifiers on the loaf volume and crumb hardness of rice breads were studied. Four different enzymes [fungal ${\alpha}$-amylase (AMYL), maltogenic bacterial ${\alpha}$-amylase (NMYL), glucose oxidases (GO), and xylanase+hemicellulases (PTP)] and four emulsifiers [sorbitan monostearate (SMS), glycerol monostearate (GMS), sodium stearoyl lactylate (SSL), and glycerol ester+propylene glycol ester+sucrose ester+sorbitan ester (SP)] were supplemented to rice dough. The addition of AMYL, GO, and GO+AMYL increased loaf volume of rice breads. The highest loaf volume was observed in rice bread supplemented with AMYL. Rice breads supplemented with enzymes firmed at lower rates during storage, and AMYL, NMYL, and GO considerably decreased crumb hardness of rice breads, exhibiting a significant antistaling effect. The addition of emulsifiers produced rice breads with better specific loaf volume and crumb texture, and continuously retarded crumb hardness of rice breads during storage. Especially, rice bread supplemented with SSL demonstrated the highest loaf volume and the lowest crumb hardness during storage.

• Microbiology and Biotechnology Letters
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• v.42 no.4
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• pp.407-412
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• 2014

A number of Saccharomycopsis fibuligera strains that can hydrolyse and utilize starch as a carbon source were isolated from nuruk, a traditional Korean starter for rice wine fermentation, and their specific growth rates on starch-containing medium were compared to choose the prominent strain. S. fibuligera strain MBY1320 showed a higher growth rate at $42^{\circ}C$ than that of strain S. fibuligera KCTC7806, indicating that S. fibuligera MBY1320 has more thermo-tolerant machinery for starch hydrolysis and utilization than KCTC7806. Although the activity of ${\alpha}$-amylase at $30^{\circ}C$ was significantly lower for S. fibuligera MBY1320 than KCTC7806 (3,812.5 U vs. 14,878.5 U), S. fibuligera MBY1320 showed a much higher glucoamylase activity at $42^{\circ}C$ than S. fibuligera KCTC7806 (5,048.9 U vs. 13,152.3 U). Thus, a new S. fibuligera strain, with a higher starch-hydrolysing activity at elevated temperatures than that of other types of strain, this study reports.

• Journal of the East Asian Society of Dietary Life
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• v.26 no.2
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• pp.163-170
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• 2016

This study investigated the quality characteristics of doenjang prepared with different Bacillus strains (Bacillus subtilis KACC 15935 and Bacillus subtilis HJ18-9). Changes in enzyme activities (protease, cellulase, and ${\alpha}-amylase$), amino-type nitrogen and ammonia-type nitrogen contents, and reducing sugar were investigated during the fermentation period. Enzymes such as protease, cellulase, and ${\alpha}-amylase$ play important roles in the composition of nutrients, as well as in the flavor and taste of doenjang. After 60 days of fermentation, protease activities in control doenjang, and doenjang fermented with B. subtilis KACC 15935, and B. subtilis HJ18-9 increased significantly up to $382.58{\pm}4.02$, $342.58{\pm}7.94$, and $392.58{\pm}1.91unit/g$, respectively (p<0.05). At the beginning of fermentation, protease activities were in the range of 156.88~182.71 unit/g. Cellulase and ${\alpha}-amylase$ activities of doenjang in HJ18-9 were higher than those in other samples. After fermentation, amino-type nitrogen in doenjang fermented with control, B. subtilis KACC 15935, and B. subtilis HJ18-9 increased significantly up to $143.25{\pm}1.62$, $141.86{\pm}2.14$, and $150.23{\pm}1.62mg%$, respectively (p<0.05). These results suggest that B. subtilis HJ18-9 is a suitable starter for the preparation of doenjang.

• The Korean Journal of Mycology
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• v.41 no.3
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• pp.160-166
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• 2013

Recovery of ${\alpha}$-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) from spent mushroom composts (SMCs) of Pleurotus cornucopiae, Pleurotus ostreatus, Pleurotus eryngii, Hericium erinaceum, Lyophyllum ulmarium, Agrocybe cylindracea, Lentinus lepideus, and Flammulina velvtipes were investigated using different extraction buffers. The maximum recovery of the enzymes was mostly detected in SMC extracts with tap water and 0.25% Triton X-100 by shaking incubation (200 rpm) for 2 h at $4^{\circ}C$. The xylanase (152 U/g) and laccase (8.1 U/g) activities were the highest in SMC extracts from F. velvtipes and P. eryngii. In addition, high enzymatic activities of ${\alpha}$-amylase (3.6 U/g) and cellulase (3.4 U/g) was detected in SMC extract of A. cylindracea. Futhermore, cellulase and laccase activities of SMCE from P. eryngii were compared to commercial enzymes.

• Applied Biological Chemistry
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• v.41 no.7
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• pp.510-515
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• 1998

Effect of freezing of soybean and addition of mugwort on the flavor development and the changes of chemical compositions in Chungkookjang was investigated. The changes in the amount of amino-type N, the activities of ${\alpha}-amylase$ and protease, and the content of volatile compounds were determined during aging of Chungkookjang. During ripening, a little higher protease activity was detected in Chungkookjang made of frozen soybean than in that made of non-frozen soybean. However, the profile of ${\alpha}-amylase$ activity of Chungkookjang made of frozen soybean was very similar to that of Chungkookjang made of non-frozen soybean. The amount of amino-type N increased gradually with the ripening period and decreased after 21 days of ripening. The amount of amino-type N was slightly higher in Chungkookjang made of frozen soybean than in that made of non-frozen soybean. Addition of mugwort had little effect on the enzyme activities and the amino-type N content. The mugwort added in Chungkookjang reduced the production of cis-3-hexenol which is responsible for the unpleasant odor, and freezing of soybean enhanced the production of 2, 6-dimethyl pyrazine which is contribute to the taste. According to sensory evaluation, flavor was the highest after the 14 days of ripening and addition of mugwort increased significantly the flavor in both Chungkookjang prepared with frozen soybean and Chungkookjang with non-frozen soybean. However, the effect of freezing of soybean on the flavor was not significant.

• Journal of Marine Life Science
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• v.7 no.2
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• pp.145-153
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• 2022

Mercury (Hg) is a major concern in marine environment because of their bioaccumulation and biomagnification properties, and adverse effects to aquatic organisms at even a trace amount. However, little information on the effects of Hg, compared to other heavy metals, is available in marine small crustaceans. Here, we investigated the transcriptional modulation of metabolism-related genes in the brackish water flea, Diaphanosoma celebensis after exposure to sublethal concentration (0.2, 0.4, 0.8 ㎍/l) of HgCl₂ for 48 h. Relative mRNA expression levels of five detoxification enzyme-coding genes (cytochrome P450; cyp360A1, cyp361A1, cyp4AP3, cyp4C122, and cyp370C5) and six digestive enzyme-coding genes [alpha amylase (AMY), alpha amylase related protein (AMY-like), trypsin (TRYP), chymotrypsin-like protein (CHY), lipase (LIP), pancreatic lipase-related protein (PLRP)] were analyzed using quantitative real time reverse transcription polymerase chain reaction (qRT-PCR). As results, Hg increased the mRNA level of cyp370C5 (clan2) and cyp4AP3 (clan4) in a concentration dependent manner. A significant increase in TRYP mRNA was also concentration-dependently observed after exposure to Hg. These findings suggest that cyp370C5 and cyp4AP3 play a key role in Hg detoxification in D. celebensis, and Hg can affect energy metabolism by modulating the transcription of digestive enzyme. This study will provide better understanding the molecular effects of Hg in marine small crustacean.