• Fisheries and Aquatic Sciences
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• v.20 no.6
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• pp.11.1-11.8
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• 2017

Juvenile black sea bream, Acanthopagrus schlegelii, were exposed to waterborne zinc (Zn) at concentrations of 0, 200, and $400{\mu}g/L$, at temperatures of 18 or $26^{\circ}C$ for 4 weeks. Superoxide dismutase (SOD) activities in the liver and gill of A. schlegelii significantly increased following exposure to waterborne Zn. Significant reduction in glutathione S-transferase (GST) activity in the liver and gill was observed following exposure to waterborne Zn. Glutathione (GSH) concentrations in the liver and gill also significantly decreased following exposure to waterborne Zn. Phagocytosis and lysozyme in the plasma and kidney were significantly increased following exposure to waterborne Zn. High water temperature increased alterations in the antioxidant and immune responses. The results of the present study suggest that waterborne Zn induced significant alterations in oxidative stress, increased immune responses and high temperature that trigger Zn toxicity.

• Molecules and Cells
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• v.23 no.2
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• pp.252-257
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• 2007

Escherichia coli HslVU is an ATP-dependent protease consisting of two heat shock proteins, the HslU ATPase and HslV peptidase. In the reconstituted enzyme, HslU stimulates the proteolytic activity of HslV by one to two orders of magnitude, while HslV increases the rate of ATP hydrolysis by HslU several-fold. Here we show that HslV alone can efficiently degrade certain unfolded proteins, such as unfolded lactalbumin and lysozyme prepared by complete reduction of disulfide bonds, but not their native forms. Furthermore, HslV alone cleaved a lactalbumin fragment sandwiched by two thioredoxin molecules, indicating that it can hydrolyze the internal peptide bonds of lactalbumin. Surprisingly, ATP inhibited the degradation of unfolded proteins by HslV. This inhibitory effect of ATP was markedly diminished by substitution of the Arg86 residue located in the apical pore of HslV with Gly, suggesting that interaction of ATP with the Arg residue blocks access of unfolded proteins to the proteolytic chamber of HslV. These results suggest that uncomplexed HslV is inactive under normal conditions, but may can degrade unfolded proteins when the ATP level is low, as it is during carbon starvation.

• Microbiology and Biotechnology Letters
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• v.22 no.2
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• pp.143-151
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• 1994

In the course of the study on strain inprovement by protoplast fusion, Lactobacillus acidophilus 88 protoplasts production and regeneration conditions were investigated. This strain produced a bacteriocin that revealed strong inhibitory activity against various indicator strains, especially L. helveticus CNRZ 1096. Protoplasts of L. acidophilus 88 strains were very efficiently obtained by treatment with 125 $\mu $g/ml lysozyme in a protoplast forming buffer containing 20 mM N-2 hydroxy-ethtl-piperazine-N'-2-ethane-sulfonic acid(HEPES, pH 7.0) and 1M sucrose at 37$\circ $C for 30 min. Hovever, treatment with mutanolysin was not effective for the production of L. acidophilus 88 protoplasts under the same conditions. High protoplast yield was obtained form the cells at the middle to late logarithmic growth phase in the de Man, Rogosa and Sharpe(MRS) medium. Regeneration was efficiently accomplished with the MRS medium containing 10% sucrose.

• Fisheries and Aquatic Sciences
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• v.15 no.3
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• pp.233-239
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• 2012

Hematological analysis can provide crucial information for monitoring the health of fish. However, there is no current information available regarding hematological changes in olive flounder following infection by Vibrio scophthalmi. In this study, hematological and biochemical alterations were determined in olive flounder infected by the high virulence strain (HVS) and low virulence strain (LVS) of V. scophthalmi. Survival in serum, skin mucus, and macrophages of olive flounder was also compared between the HVS and LVS. The results demonstrated that the hematocrit value in infected fish declined from 23.4% at 0 h to 18.0% at 168 h post infection. The total protein concentration in fish infected with the HVS was significantly higher than in fish infected with the LVS and a non-infected control. Lysozyme activity was significantly different between infected and control fish. The HVS survived in serum and cell numbers increased substantially, while cell numbers of the LVS in serum decreased. These changes in hematological characteristics in fish infected by V. scophthalmi can be used as an effective and sensitive index to monitor the physiological and pathological conditions of fish. The survival and reproduction of V. scophthalmi in host serum, skin mucus, and macrophages play a major role in systemic infection and can serve as a virulence indicator for different strains.

• YAKHAK HOEJI
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• v.37 no.2
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• pp.149-157
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• 1993

Protein methylase inhibitor which is a modulator of biological methylation has been purified and characterized from porcine liver soluble fraction by cell fractionation, Sephadex G25 chromatography, reverse phase HPLC, size exclusion HPLC. The results are summarized as follows. 1) The purified inhibitor shows apparent homogeneity, as judged by HPLC. 2) A molecular weight of the purified inhibitor which is composed of 18 amino acid residues is about 1,400 daltons. 3) A single absorption peak of ultraviolet spectrum was observed at 260nm. 4) The inhibitor was not inactivated by heating at $100^{\circ}C$ until 60min. and its activity was not influenced by treatment with digestive enzymes, such as trypsin, pepsin, pronase, chymotrypin, lysozyme, DNase, and RNase. 5) The purified inhibitor inhibited protein rnethylase I, II, III and phospholipid methyltransferase activities. 6) The purified inhibitor inhibited noncompetitively protein methylase II from porcine liver, spleen, and testis. 7) The $K_{i}$ values for protein methylase II from porcine liver, spleen, and testis were 300nM, 250nM, 297nM, respectively.

• Bulletin of the Korean Chemical Society
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• v.25 no.12
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• pp.1917-1923
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• 2004

Although the proteolytic activity of the Cu(II) complex of cyclen (Cyc) is greatly enhanced upon attachment to a cross-linked polystyrene (PS), the Cu(II)Cyc-containing PS derivatives reported previously hydrolyzed only a very limited number of proteins. The PS-based artificial metalloproteases can overcome thermal, mechanical, and chemical instabilities of natural proteases, but the narrow substrate selectivity of the artificial metalloproteases limits their industrial application. In the present study, artificial metalloproteases exhibiting broad substrate selectivity were synthesized by attaching Cu(II)Cyc to a PS derivative using linkers with various structures in an attempt to facilitate the interaction of various protein substrates with the PS surface. The new artificial metalloproteases hydrolyzed all of the four protein substrates (albumin, myoglobin, ${\gamma}$-globulin, and lysozyme) examined, manifesting $k_{cat}/K_m$ values of 28-1500 $h_{-1}M_{-1}$ at 50 $^{\circ}C$. The improvement in substrate selectivity is attributed to steric and/or polar interaction between the bound protein and the PS surface as well as the hydrophobicity of the microenvironment of the catalytic centers.

• Journal of Microbiology and Biotechnology
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• v.18 no.5
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• pp.946-951
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• 2008

The opd gene, encoding organophosphorus hydrolase (OPH) from Flavobacterium sp. capable of degrading a wide range of organophosphate pesticides, was surface- and intracellular-expressed in Synechococcus PCC7942, a prime example of photoautotrophic cyanobacteria. OPH was displayed on the cyanobacterial cell surface using the truncated ice nucleation protein as an anchoring motif. A minor fraction of OPH was displayed onto the outermost surface of cyanobacterial cells, as verified by immunostaining visualized under confocal laser scanning microscopy and OPH activity analysis; however, a substantial fraction of OPH was buried in the cell wall, as demonstrated by proteinase K and lysozyme treatments. The cyanobacterial outer membrane acts as a substrate (paraoxon) diffusion barrier affecting whole-cell biodegradation efficiency. After freeze-thaw treatment, permeabilized whole cells with intracellular-expressed OPH exhibited 14-fold higher bioconversion efficiency ($V_{max}/K_m$) than that of cells with surface-expressed OPH. As cyanobacteria have simple growth requirements and are inexpensive to maintain, expression of OPH in cyanobacteria may lead to the development of a low-cost and low-maintenance biocatalyst that is useful for detoxification of organophosphate pesticides.

• Journal of Microbiology and Biotechnology
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• v.32 no.9
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• pp.1209-1216
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• 2022

To better understand the effects of PEGylation and biotinylation on the delivery efficiency of proteins, the cationic protein lysozyme (LZ) and anionic protein bovine serum albumin (BSA) were chemically conjugated with poly(ethylene glycol) (PEG) and biotin-PEG to primary amine groups of proteins using N-hydroxysuccinimide reactions. Four types of protein conjugates were successfully prepared: PEGylated LZ (PEG-LZ), PEGylated BSA (PEG-BSA), biotin-PEG-conjugated LZ (Bio-PEG-LZ), and biotin-PEG-conjugated BSA (Bio-PEG-BSA). PEG-LZ and Bio-PEG-LZ exhibited a lower intracellular uptake than that of LZ in A549 human lung cancer cells (in a two-dimensional culture). However, Bio-PEG-BSA showed significantly improved intracellular delivery as compared to that of PEG-BSA and BSA, probably because of favorable interactions with cells via biotin receptors. For A549/fibroblast coculture spheroids, PEG-LZ and PEG-BSA exhibited significantly decreased tissue penetration as compared with that of unmodified proteins. However, Bio-PEG-BSA showed tissue penetration comparable to that of unmodified BSA. In addition, citraconlyated LZ (Cit-LZ) showed reduced spheroid penetration as compared to that of LZ, probably owing to a decrease in protein charge. Taken together, chemical conjugation of targeting ligands-PEG to anionic proteins could be a promising strategy to improve intracellular delivery and in vivo activity, whereas modifications of cationic proteins should be more delicately designed.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.41 no.4
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• pp.248-252
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• 2008

A 6-week feeding trial was conducted to investigate effects of dietary supplementation with wood vinegar (WV) as a feed additive for juvenile olive flounder, Paralichthys olivaceus. Eight experimental diets supplemented with 0(control), 0.01, 0.025, 0.05, 0.1, 0.25, 0.5 and 2.0% WV/kg diet($WV_0,\;WV_{0.01},\;WV_{0.025},\;WV_{0.05},\;WV_{0.1},\;WV_{0.25},\;WV_{0.5},\;and\;WV_{2.0}$, respectively) were prepared by mixing with basal diet. After the 6-week feeding trial, fish fed the $WV_{0.1}$ diet had significantly higher weight gain, feed efficiency, and specific growth rate than those of fish fed the other diets(P<0.05). No mortality was observed in the treatment groups fed the experimental diets for 6 weeks. The liver antioxidant activity of fish fed the $WV_{0.01},\;WV_{0.025},\;and\;WV_{0.05}$ diets was significantly higher than that of fish fed the other diets. The lysozyme activity of fish fed the $WV_{0.25}$ diet was significantly higher than that of fish fed the other diets. Fish fed diets supplemented with WV had significantly lower cumulative mortality than that of fish fed the control diet throughout the challenge test, and fish fed the $WV_{0.05}\;and\;WV_{0.1}$ diets had the highest resistance to disease caused by Edwardsiella tarda at day 7 of the challenge test. Therefore, feeding 0.05-0.1% WV/kg diet improves the growth and immune responses of juvenile olive flounder, Paralichthys olivaceus.

• Journal of fish pathology
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• v.32 no.2
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• pp.91-96
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• 2019

Rainbow trout (Oncorhynchus mykiss) were treated with ultrasonic (40kHz) and microbubble. The organic serum components, glucose were decreased compared to control at microbubble single treatment (M) and ultrasonic treatment (UL), but glucose concentrations in fish serum of all experimental groups analyzed in this study were also within the normal range. The organic serum components, cholesterol and total protein were no significant change by ultrasonic and microbubble treatment. There were no significant changes in the lysozyme activity on ultrasonic and microbubble treatment. Serum bactericidal activity decreased on ultrasonic single treatment (UL) compared to the control (P<0.05), but the numbers of bacterial colonies were significantly lower than those in the negative (PBS). The results of this study indicate that ultrasonic and microbubble treatment did not have a critical effect on the serum composition and innate immunity of O. mykiss, and that it is a way to do not significant stress on the fish body as a countermeasure against sea lice.