• Korean Journal of Fisheries and Aquatic Sciences
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• v.42 no.5
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• pp.426-433
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• 2009

Fish scales have potential in functional food preparation due to their antioxidant and antihypertensive properties. We investigated the angiotensin I converting enzyme (ACE) inhibitory activity of Tilapia mossambica scale extracts. Hydrolysates of tilapia scales were prepared by enzymatic extraction using five proteases (${\alpha}$-chymotrypsin, Alcalase, Kojizyme, Protamex and trypsin) after scales were treated with hot water for 3 hr. Scale enzymatic hydrolysates prepared using both hot water and enzyme treatments exhibited elevated hydrolysis (about 25%-55%) compared to only enzyme treatment (about 15%-45%). Enzymatic hydrolysates (1 mg/mL) prepared by both hot water and enzyme treatments also showed significantly increased ACE inhibitory activities from about 20%-75%. The pattern of ACE inhibitory activities was similar to the degree of hydrolysis. Alcalase and ${\alpha}$-chymotrypsin hydrolysates displayed the highest ACE inhibitory activities ($IC_{50}$ = 0.83 mg/mL and 0.68 mg/mL, respectively). In addition, the ACE inhibitory effects of $IC_{50}$-chymotrypsin hydrolysates increased with decreasing molecular weight (5 kDa>, 10 kDa> and 30 kDa>), with the 5 kDa> fraction displaying the highest ACE inhibitory activity (about 89.9% and $IC_{50}$ = 0.1 mg/mL). We suggest that the peptide compounds of enzymatic hydrolysates prepared from tilapia scale enhances ACE inhibitory activity and might be useful as an antihypertensive material.

• Korean Journal of Agricultural Science
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• v.39 no.4
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• pp.561-568
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• 2012

The aim of this study was to introduce a simple method for isolation of ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin and bovine serum albumin from cow's milk, and peptides produced by enzymatic hydrolysis of ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin and bovine serum albumin with alcalase. Whey protein were precipitated from whey by ammonium sulfate and, ${\alpha}$-lactalbumin and ${\beta}$-lactoglobulin were isolated using Hi Prep 26/60 Sephacryl S-100 column gel filtration chromatography. Bovine serum albumin and ${\beta}$-lactoglobulin were isolated by Mono-Q 5/50 GL column anion exchange chromatography of the 50% Ammonium Sulfate-supernatant. Isolated whey proteins were hydrolyzed by proteolytic alcalase. Tricine SDS-PAGE and reverse-phase HPLC analyses revealed that almost hydrolyzed all the ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin and bovine serum albumin with alcalase. Molecular weight of various peptides derived from alcalase hydrolysate were small molecular weight than 3.5 kDa.

• Asian-Australasian Journal of Animal Sciences
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• v.10 no.4
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• pp.398-401
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• 1997

A caecal fermentation study was conducted in 30 Angora rabbits equally placed under five whole diets (75 concentrate : 25 roughage) supplemented with processed Neem (Azadirachta indica) kernel meal (NKM ; 2% urea or 1.5% NaOH, W/W), replacing isonitrogenously either at 50 or 100% level of deoiled peanut (Arachis hypogea) meal, for 18 wk period. The total volatile fatty acids level was depressed (p < 0.05) in all the experimental groups (0.02 to 0.04 mEq/g) and ammonia-nitrogen was lowest (p < 0.05; $24.9{\mu}mol/g$) in rabbits fed NaOH treated NKM supplemented diet. Enzyme activity (unit/g) of carboxy methyl-cellulase, ${\alpha}$-amylase, protease and urease exhibited much variation and did not differ significantly. Hence, the results could not confirm a possible adverse effect of feeding NKM on caecal fermentation.

• Toxicological Research
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• v.18 no.3
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• pp.275-283
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• 2002

Fungal metabolite, gliotoxin is an epipolythiodioxopiperazin (ETP) class and has various roles including immunomodulatory and apoptotic effects. This study was designed to evaluate the mechanism by which gliotoxin exerts the apoptosis on human promyelocytic leukemic HL-60 cells. Herein, we demonstrated that the gliotoxin decreased the cell viability in a time-dependent manner Gliotoxin-induced cell death was confirmed us apoptosis characterized by chromatin condensation and ladder-pattern fragmentation of genomic DNA. Gliotoxin increased the catalytic activities of caspase-3 and caspase-9. Activation of caspase-3 was further confirmed by degradation of procaspase-3 and poly(ADP-ribose) polymerase (PARP) by gliotoxin in HL-60 cells. Furthermore, gliotoxin induced the changes of mitochondrial transmembrane potential (MTP). Antioxidants, including GSH and NAC, markedly inhibited apoptosis with conistent suppression of enzymatic activity of caspase-3, caspase-9, and MTP loss in gliotoxin-treated cells. Taken together, we suggest that gliotoxin function as an oxidant and ploys proapoptotic roles in HL-60 cells via activation of intrinsic caspase cascades as well as mitochondrial dysfunction.

• Bulletin of the Korean Chemical Society
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• v.4 no.2
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• pp.72-76
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• 1983

Cephaloglycin was synthesized directly from D-${\alpha}$ -phenylglycine methyl ester and 7-aminocephalosporanic acid using whole cell enzyme of Xanthomonas citri (IFO 3835). Some optimal conditions for cephaloglycin synthesis were investigated, and yield improvements for its production by several methods were attempted. Using the whole cell enzyme system, the reaction kinetic model for cephaloglycin synthesis is proposed, and the kinetic constants for D-${\alpha}$ -phenylglycine methyl ester hydrolysis, cephaloglycin synthesis, and cephaloglycin hydrolysis were determined. The $K_m$ values of D-${\alpha}$-phenylglycine methyl ester, 7-aminocephalosporanic acid, and cephaloglycin were 11 mM, 24 mM, and 167 mM, and $K_i$ value of D-${\alpha}$-phenylglycine was 15 mM, respectively. The pattern of product inhibition was found to be competitive one.

• The korean journal of orthodontics
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• v.54 no.4
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• pp.199-209
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• 2024

The clinical application of aligners is accompanied by the ageing of the polymer appliances and the attachments used, which may result in inefficiency in reaching the predicted range of tooth movement, and release of compounds and microplastics in the oral cavity as a result of the friction, wear and attrition of the aligner and composite attachment. The purpose of this review is to present the mechanism and effects of in vivo ageing; describe the hydrolytic degradation of aligners and enzymatic degradation of composite attachments; examine the ageing pattern of aligners in vivo, under actual clinical scenarios; and identify a link to the discrepancy between predicted and actual clinical outcome. Lastly, strategies to deal with three potentially critical issues associated with the use of aligners, namely the necessity of weekly renewal, the dissimilar mechanical properties of aligner and attachment resulting in wear and plastic deformation of the aligner, and the development of integuments and biofilms with microbial colonization of the appliance, are discussed.

• Journal of Microbiology and Biotechnology
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• v.8 no.2
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• pp.152-157
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• 1998

To analyze the role of the C and D domains in the cyclization activity of cyclodextrin glucanotransferase (CGTase), two plasmids, pKB1ΔC300 and pKB1ΔD96, were constructed in which DNA regions encoding 100 and 32 amino acids, respectively, from the C and D domains of B. stearothermophilus NO2 CGTase were deleted. The mutated CGTase from the pKBlΔC300 produced much lower amounts of ${\alpha}$-, ${\beta}$-, and $\gamma$-cyclodextrin (CD) than the parental CGTase. However, the mutated CGTase from the pKBlΔD96 showed a similar production pattern of CDs to wild-type CGTase. The production ratios of the ${\alpha}$-, ${\beta}$- and $\gamma$-CDs were not affected by the deletions, when compared to those of parental CGTase. The optimum temperature of the mutated CGTase from the pKBlΔC300 was decreased from $60^{\circ}C$ to $55^{\circ}C$. The optimum pH of the mutated CGTase from the pKB1D96 was shifted from 6.0 to 7.0. The thermostability of the two mutant CGTases were not changed. From these results, it is suggested that the C and D domains are not related to cyclization activity directly because mutant-enzymes deleted C or D domains still possessed their activity. However, they are important for other enzymatic properties such as productivity and pH optimum as a partition of CGTase tertiary structure.

• BMB Reports
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• v.38 no.5
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• pp.624-631
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• 2005

Tuberculosis, caused by Mycobacterium tuberculosis, continues to be one of the main diseases to mankind. It is urgent to discover novel drug targets for appropriate antimicrobial agents against this human pathogen. The shikimate pathway is onsidered as an attractive target for the discovery of novel antibiotics for its essentiality in bacteria and absence in mammalian cells. The Mycobacterium tuberculosis aroE-encoded shikimate dehydrogenase was cloned, expressed and purified. Sequence alignment analysis shows that shikimate dehydrogenase of Mycobacterium tuberculosis exhibit the pattern of G-X-(N/S)-V-(T/S)-X-PX-K, which is highly conserved within the shikimate dehydrogenase family. The recombinant shikimate dehydrogenase spectrum determined by CD spectroscopy showed that the percentages for $\alpha$-helix, $\beta$-sheet, $\beta$-turn, and random coil were 29.2%, 9.3%, 32.7%, and 28.8%, respectively. The enzymatic characterization demonstrates that it appears to be fully active at pH from 9.0 to 12, and temperature $63^{\circ}C$. The apparent Michaelis constant for shikimic acid and $NADP^+$ were calculated to be about $29.5\;{\mu}M$ and $63\;{\mu}M$. The recombinant shikimate dehydrogenase catalyzes the substrate in the presence of $NADP^+$ with an enzyme turnover number of $399\;s^{-1}$. Zymological studies suggest that the cloned shikimate dehydrogenase from M. tuberculosis has a pretty activity, and the work should help in the discovery of enzyme inhibitors and further of possible antimicrobial agents against Mycobacterium tuberculosis.

• Journal of Life Science
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• v.24 no.12
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• pp.1378-1382
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• 2014

Phospholipase D (PLD) catalyzes the hydrolysis of phospholipid to phosphatidic acid (PA), a lipid secondary messenger. Two forms of PLD isozymes, phosphatidylcholine-specific PLD1 and PLD2, have been identified. PLD has emerged as a critical regulator of cell proliferation and survival signaling, and dysregulation of PLD occurs in a various illnesses, including cancer. PLD activity is essential for cell survival and protection from apoptosis. Overexpression of PLD isozymes or PLD-generated PA attenuates the expression of apoptotic genes and confers resistance to apoptosis. The apoptosis-related molecular mechanisms of PLD remain largely unknown. Recently, the dynamics of PLD turnover during apoptosis have been reported. The cleavage of PLD isozymes as specific substrates of caspase differentially regulates apoptosis. PLD1 is cleaved at one internal site, and PLD2 is cleaved two sites at the front of the N-terminus. The cleavage of PLD1 reduces its enzymatic activity, probably via the dissociation of two catalytic motifs, whereas the cleavage of PLD2 does not affect the catalytic motifs and its activity. Thus, PLD2 maintains antiapoptotic capacity, despite its cleavage. Therefore, the differential cleavage pattern of PLD isozymes by caspase affects its enzymatic activity and antiapoptotic function. Thus, PLD is considered a potential target for cancer therapy. We summarize recent studies regarding the functional role of PLD in apoptosis.

• Journal of Life Science
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• v.11 no.6
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• pp.509-516
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• 2001

Enzymes and non-enzymatic antioxidants are involved in defense of oxgen free radical intermediates in all aerobic cells. The non -enzymatic antioxidants and antioxidant enzyme from the extracts of Solanum nigrum L. known to be anticancer medicinal plant were examined in other to utilize the discovery in natural products as cancer chem-opereventive agents. The DPPH(1,1-diphenyl-2-picryl-hydrazyl) free radical scavening activity on plant position of Solanum nigrum L. was the highest in root, with stem, whole plant, seed, leaf and flower, at higher activities respectively. In extraction methods, the DPPH free radical scavenging activity by circulating extraction with 80 % MeOH. The DPPH activity of L6 fraction by LH-20 column chromatography showed about 6.7 times higher than that of ethyl acetate-fraction. These were identified as phenolic compounds such as 2-6-methano-3-benzazocin-11-ol, 2[1H]-phyidinethione and 2-hydroxy -5-methyl-benzaldehyde. Peroxidase(POD) and superoxide dismutase(SOD) activities of stem and root were higher than that of other plant positions and those of plant positions according to growing stage were the highest in 60 days after seeding. The numbers of isozyme pattern of POD and SOD showed 10 hands and 5 bands, respectively, especially, 8 bands of POD and 3 bands of SOC showed a difference according to plant positions.