• Asian-Australasian Journal of Animal Sciences
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• v.31 no.10
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• pp.1627-1634
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• 2018

Objective: The study aimed to evaluate the effect of replacing wheat bran for cactus cladodes plus urea (0%, 25%, 50%, 75%, and 100%) on the intake of nutrients, nitrogen balance, microbial protein synthesis, and rumen fermentation for steers. Methods: Five crossbred steers (1/2 Holstein-Zebu), with rumen cannula and an average body weight of $180{\pm}5.3kg$, were assigned to a $5{\times}5$ Latin square design. Dietary treatments consisted of the replacement of the total of wheat bran in basal diet by cactus cladodes using the following proportions: 0% for basal diet, 25%, 50%, 75%, and 100% cactus cladodes replacing wheat bran. Urea was added to the diets to adjust the crude protein (CP) content to 130 g/kg dry matter. Results: Maximum dry matter intake (5.73 kg/d) and maximum nitrogen balance (103 g/d) were estimated for 54.6% and 70.8% replacement levels of wheat bran. The maximum microbial protein production (44.6 g/d) was obtained at a replacement level of 49.7%, and a medium value (125 g CP mic/kg total digestible nutrients) of microbial protein efficiency was observed. The rumen pH increased linearly according to cactus cladodes inclusion, while the ammonia nitrogen medium value was 24.5 mg/dL. Conclusion: The replacement of 55% wheat bran for cactus cladodes plus urea in the diet of crossbred steers is recommended.

• The Journal of Internal Korean Medicine
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• v.29 no.2
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• pp.469-489
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• 2008

Objectives : The purpose of this study was to investigate the effects of Chungganhaeju-tang(Qingganjiejiu-tang) on alcoholic liver damaged by applying proteomics. Materials and Methods : Sprague-Dawley rats were used in this experiment the rats were divided into the normal group, the control group(alcohol) and the sample group(CGHJT +alcohol). The ethanol was orally administered twice a day for 6 weeks in the control and sample groups. Water instead of ethanol was orally administered twice a day for 6 weeks in the normal group. CGHJT extract was orally administered once a day for 6 weeks in the sample group. The livers of each group were processed and assessed by histology, Western Blot, $Oxyblot^{TM}$, CBB and 2-dimensional electrophoresis. Results : In the histological findings of the liver, CGHJT inhibited hepatic fibrogenesis induced by alcohol. TIMP-1 decreased in the sample group assessed by western blot and statistical significance was noted by dot blotting(p<0.05). In the $Oxyblot^{TM}$, protein oxidation induced by alcohol treatment decreased with CGHJT. In the 2-dimensional electrophoresis finding, increased proteins alcohol such as HSP 60, 60kDa heat shock protein, 3-mercaptopyruvate sulfurtransferase were normalized by CGHJT. CGHJT was considered to normalize the anti-oxidation activity elevated by alcohol. In the 2-dimensional electrophoresis finding, increased oxidized proteins such as actin, prolyl 4-hydroxylase beta polypeptide, 94kDa glucose regulated protein(GRP94), heat shock protein 90-alpha(HSC86), calreticulin precursor(CRP55), ATP synthase beta chain mitochondrial precursor, caspase-8 precursor, and dihydrolipoamide succinyltransferase(E2) decreased with CGHJT. CGHJT was considered to reduce the oxidative stress of alcohol. Conclusion : Chungganhaeju-tang(Qingganjiejiu-tang) exerts an inhibitory effect against the fibrosis and protein oxidation induced by alcohol treatment of rat liver. CGHJT was considered to normalize the elevated anti-oxidation activity by alcohol and to reduce the level of oxidative stress due to alcohol.

• Fisheries and Aquatic Sciences
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• v.5 no.1
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• pp.21-27
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• 2002

Angiotensin I converting enzyme inhibitory activities of shelled krill (Euphausia superba) hydrolysates by autolysis and by hydrolysis with commercial proteases were analyzed. Among the proteases, Alcalase was the most effective protease for the hydrolysis of krill considering the degree of hydrolysis $(87.5\%)$ and the ACE inhibitory activity $(60\%)$. Four hour hydrolysis suggested as the most suitable and economic. In order to establish the optimum hydrolysis condition of krill, degree of hydrolysis and ACE inhibitory activity as affected by Alcalase concentration and water amount added were statistically analyzed by response surface methodology (RSM). The optimum hydrolysis condition was $2.0\%$ Alcalase hydrolysis in 2 volumes (v/w) of water at $55\% for 4 hr. The hydrolysate prepared from the optimum hydrolysis condition was fractionated by molecular weight. The lower molecular weight fraction showed the higher ACE inhibitory activity. $IC_{50}$ of the fraction under 500 Da was 0.57mg protein/mL.

• Applied Microscopy
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• v.48 no.3
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• pp.55-61
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• 2018

The synaptic vesicle is a specialized structure in presynaptic terminals that stores various neurotransmitters. The actin filament has been proposed for playing an important role in mobilizing synaptic vesicles. To understand the role of actin filament on synaptic vesicle pooling, we characterized synaptic vesicles and actin filament after treatment of brain-derived neurotrophic factor (BDNF) or Latrunculin A on primary cultured neuron from rat embryo hippocampus. Western blots revealed that BDNF treatment increased the expression of synapsin I protein, but Latrunculin A treatment decreased the synapsin I protein expression. The increased expression of synapsin I after BDNF disappeared by the treatment of Latrunculin A. Three-dimensional (3D) tomography of synapse showed that more synaptic vesicles localized near the active zone and total number of synaptic vesicles increased after treatment of BDNF. But the number of synaptic vesicle was 2.5-fold reduced in presynaptic terminals and the loss of filamentous network was observed after Latrunculin A application. The treatment of Latruculin A after preincubation of BDNF group showed that synaptic vesicle number was similar to that of control group, but filamentous structures were not restored. These data suggest that the actin filament plays a significant role in synaptic vesicles pooling in presynaptic terminals.

• Journal of Microbiology and Biotechnology
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• v.18 no.1
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• pp.160-166
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• 2008

A DNA fragment containing 2,079 base pairs from Bacillus circulans CGMCC 1416 was cloned using degenerate PCR and inverse PCR. An open reading frame containing 981 bp was identified that encoding 326 amino acids residues, including a putative signal peptide of 31 residues. The deduced amino acid sequence showed the highest identity (68.1%) with $endo-{\beta}-1,4-D-mannanase$ from Bacillus circulans strain K-1 of the glycoside hydrolase family 5 (GH5). The sequence encoding the mature protein was cloned into the pET-22b(+) vector and expressed in Escherichia coli as a recombinant fusion protein containing an N-terminal hexahistidine sequence. The fusion protein was purified by $Ni^{2+}$ affinity chromatography and its hexahistidine tag cleaved to yield a 31-kDa ${\beta}$-mannanase having a specific activity of 481.55U/mg. The optimal activity of the purified protein, MANB48, was at $58^{\circ}C$ and pH 7.6. The hydrolysis product on substrate locust bean gum included a monosaccharide and mainly oligosaccharides. The recombinant MANB48 may be of potential use in the feed industry.

• Fisheries and Aquatic Sciences
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• v.25 no.7
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• pp.390-402
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• 2022

In this study we investigate physical and chemical characteristics of immature and mature skipjack tuna (Katsuwonus pelamis) roes in fresh and dried forms. Fresh roes were studied for histological structure and also dried by three methods: hot air drying (HD), vacuum drying (VD) and freeze drying (FD). The obtained roe powders were analysed for proximate composition, color value, fatty acid composition, amino acid profile, equivalent umami concentration (EUC) and protein pattern. Unyolked oocytes were more common in immature roes, while fully yolked oocytes were more common in mature roes. All dried tuna roes contained high content of protein and lipid (69.31%-70.55% and 11.14%-16.02%, respectively). The powders obtained by FD provided the highest lightness value (L^*). The main fatty acid found in all roe powders was docosahexaenoic acid (DHA) (23.49%-27.02%). Glutamic acid, leucine, and aspartic acid were the three most abundant amino acids found in the powders (13.58-14.61, 8.06-8.42, and 7.81-8.39 g/100 g of protein, respectively). The mature roe powder obtained from HD provided the highest EUC value (73.09 g monosodium glutamate/100 g of samples). The protein band at molecular weight of 97 kDa (vitelline) represented the major protein. Therefore, dried tuna roe could be a functional ingredient source of protein and lipid rich in DHA and it also has potential to be used as taste enhancer with umami compound.

• Molecules and Cells
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• v.24 no.2
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• pp.261-267
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• 2007

Apoptotic signals are typically accompanied by activation of aspartate-specific cysteine proteases called caspases, and caspase-3 and -7 play crucial roles in the execution of apoptosis. Previously, using the proteomic approach, protein disulfide isomerase (PDI) was found to be a candidate substrate of caspase-7. This abundant 55 kDa protein introduces disulfide bonds into proteins (via its oxidase activity) and catalyzes the rearrangement of incorrect disulfide bonds (via its isomerase activity). PDI is abundant in the ER but is also found in non-ER locations. In this study we demonstrated that PDI is cleaved by caspase-3 and -7 in vitro. In addition, in vivo experiment showed that it is cleaved during etoposide-induced apoptosis in HL-60 cells. Subcellular fractionation showed that PDI was also present in the cytosol. Furthermore, only cytosolic PDI was clearly digested by caspase-3 and -7. It was also confirmed by confocal image analysis that PDI and caspase-7 partially co-localize in both resting and apoptotic MCF-7 cells. Overexpression of cytosolic PDI (ER retention sequence deleted) inhibited cell death after an apoptotic stimulus. These data indicate that cytosolic PDI is a substrate of caspase-3 and -7, and that it has an anti-apoptotic action.

• Microbiology and Biotechnology Letters
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• v.36 no.3
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• pp.182-188
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• 2008

A hyperthermophilic bacteria (strain HJ6) was isolated from a hot springs located in the Arima-cho, Hyogo, Japan. The cells were long-rod type ($2-4{\mu}m$), about $0.4{\mu}m$ in diameter. The pH and temperature for optimal growth were 6.5 and $80^{\circ}C$, respectively. Phylogenetic analysis based on the 16S rDNA sequence and biochemical studies indicated that HJ6 belonged to the genus Thermus thermophilus (Tt). The gene encoding the Trehalose synthase (TS) was cloned and sequenced. The open reading frame (ORF) of the TtTS gene was composed of 2,898 nucleotides and encoded a protein (975 amino acids) with a predicted molecular weight of 110.56 kDa. The deduced amino acid sequence of TtTS showed 99% and 83% identities to the Thermus caldophilus TS and Meiothermus ruber TS, respectively. TtTS gene was expressed in Escherichia coli cells, and the recombinant protein was purified to homogeneity. The optimal temperature and pH for Trehalose synthase activity were found to be $80^{\circ}C$ and 7.5, respectively. The half-life of heat inactivation was about 40 min at $90^{\circ}C$. The maximum trehalose conversion rate of maltose into trehalose by the enzyme increased as the substrate concentration increased, and reached 55.7% at the maltose concentration of 500 mM, implying that the enzyme conversion was dependent of the substrate concentration.

• Journal of the Korean Applied Science and Technology
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• v.35 no.2
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• pp.367-377
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• 2018

Swimming crab(Ovalipes punctatus) is produced in Korea and utilized as semi-processed food at streamed cooked state. Recently, protein hydrolysates have been known as having function such as antioxidant, suppression of hypertension, immunodulatory, alleviation of pain, and antimicrobial activity. This research was investigated to find the functional antioxidant from crab hydrolysates. To fine optimal protease enzyme, alcalase, bromelain, flavourzyme, neutrase, papain, and protamex were selected to evaluate the DPPH radical scavenging activity and finally bromelain to show the best activity was selected. The molecular weight of bromelain hydrolysates were distributed with range from 500 to 3,200 Da and 7 different molecules or more. The amino acids related to antioxidant capacity was about 42.54%. The processes optimization study used was the response surface methodology. The ranges of processes were the reaction temperature of 40 to $60^{\circ}C$, pH 6 to 8, and enzyme concentration 1 to 3%(w/v). As a result, the optimization of process was determined at temperature of $55^{\circ}C$, pH of 6.5, and enzyme concentration of 3%(w/v). In these conditions, degree of hydrolysates were maximum 71.60%. Therefore, we expect that those products are useful as functional food ingredients.

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1077-1084
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• 2009

A genomic library was constructed to clone a xylanase gene (Mxyn10) from Demequina sp. JK4 isolated from a deep sea. Mxyn10 encoded a 471 residue protein with a calculated molecular mass of 49 kDa. This protein showed the highest sequence identity (70%) with the xylanase from Streptomyces lividans. Mxyn10 contains a catalytic domain that belongs to the glycoside hydrolase family 10 (GH10) and a carbohydrate-binding module (CBM) belonging to family 2. The optimum pH and temperature for enzymatic activity were pH 5.5 and $55^{\circ}C$, respectively. Mxyn10 exhibited good pH stability, remaining stable after treatment with buffers ranging from pH 3.5 to 10.0. The protein was not significantly affected by a variety of chemical reagents, including some compounds that usually inhibit the activity of other related enzymes. In addition, Mxyn10 showed activity on cellulose. These properties mark Mxyn10 as a potential enzyme for industrial application and saccharification processes essential for bioethanol production.