• Microbiology and Biotechnology Letters
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• v.29 no.2
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• pp.72-77
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• 2001

Expression of f3 ~agarase Gene and Catabolite Repression in Escherichia coli by the Promoter of Alginate Lyase Gene Isolated from Marine Pseudomonas sp. Jin, Cheal~Ho, J~Hyeon Park, Jeong-Hyun Han, YoonM Hyeok Chae, Jong~Hee Lee, Jung-Kee Lee!, and In-800 Kong*. Faculty of Food Science and Biotechnology, Pukyong National UniversitYt Pusan 608-737, Korea, llnBioNet Co. 1690-3 Taejon 306-230, Korea - Promoter is a key factor for expression of the recombinant protein. There are many promoters for overexpression of protein in various organisms. The aly promoter of Pseudomonas sp. W7 isolated from marine environment was known to be a constitutive expression promoter of the alginate lyase gene, and it's promoter activity is repressed by glucose in Escherichia coli. To investigate the catabolite repression of the aly promoter ~md association between the promoter mutants, f3 agarase gene, which was also cloned from Pseudomonas sp. W7 was connected to the aly promoter with the sequence the coding 46 N-terminal amino acids ofthe alginate lyase gene. The constructed plasmid was introduced into E. coli and the agarase activity was measured. Fourty six amino acids of the alginate lyase gene was serially deleted using peR to the direction of 5' upstream region and subcloned. The agarase was overexpressed by the aly promoter and the production of agarase was repressed by the addition of glucose into culture media. Fourty six amino acids of alginate lyase did not affect the production of agarase at all. The deletion of a putative stem-loop structure in the aly promoter induced the decrease of f3 -agarase productivity.

• Microbiology and Biotechnology Letters
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• v.40 no.2
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• pp.92-97
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• 2012

Antimicrobial peptides (AMPs) are important components of living organisms acting against Gram-negative and Gram-positive bacterial and fungal pathogens. Cathelicidin human peptides have a variety of biological activities that can be used in clinical applications. AMPs are not produced naturally in large quantities, and chemical synthesis is also economically impractical, especially for long peptides. Therefore, as an alternative, heterologous expression of AMPs by recombinant techniques has been studied as a means to reduce production costs. E. coli is an excellent host for the expression of AMPs, as well as other recombinant proteins, because of the low cost involved and its easy manipulation. However, overexpression of AMPs in E. coli has been shown to cause difficulties resulting from the toxicity of the subsequently produced AMPs. Therefore, fusion expression was theorized to be a solution to this problem. In this study, AMPs were expressed as fused proteins with the glutathione S-transferase (GST) binding protein to protect against the toxicity of AMPs when expressed in E. coli. The LL37, and hybrid gaegurin and LL37 (GGN4(1-16)-LL37(17-32), which we designated as GL32, peptides were expressed as GST-fusion proteins in E. coli and the fusion proteins were then purified by affinity columns. The purified peptides were obtained by removal of GST and were confirmed by western blot analysis. The purified antimicrobial peptides then demonstrated antimicrobial activities against Gram-negative and Gram-positive bacterial strains.

• Ocean and Polar Research
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• v.30 no.3
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• pp.303-312
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• 2008

Despite the impacts of the climate changes on the pelagic ecosystem, few studies have examined the pelagic-benthic coupling in the adjacent East China Sea and Korea Strait. Therefore, the species composition and abundance of the macrobenthic community, as well as the potential food sources of benthic fauna were investigated in the present study using stable isotope analysis (${\delta}^{13}C\;and\;{\delta}^{15}N$) for suspended particulate organic matter (SPOM), sedimentary organic matter (SOM), phytoplankton, and zooplankton. A total of 157 macrobenthic fauna were collected, and the density of the macrobenthic fauna ranged from 4 to 434 ind./0.25 $m^2$, with an average density of 149 ind./0.25 $m^2$. The density of the benthic fauna increased moving from offshore shelf sites to coastal sites adjacent to the Korea Strait. Cluster analysis showed that the macrobenthic communities consisted of three distinct groups: group A in the Korea Strait, group B in the East China Sea, and group C near Ieodo. The dominant species in group A were the amphipods Photis japonica and Ampelisca miharaensis, followed by the polychaete Scolotoma longifolia. Environmental variables, such as the temperature of the seawater and sediment, and oxygen, and chlorophyll a levels, appeared to affect the structure of the community, suggesting the importance of coupling with the pelagic system. The ${\delta}^{13}C$ values of SPOM and zooplankton ranged from -22.97 to -23.5% and -19.92 to -21.86%, respectively, showing a relatively narrow range(<1%) between the two components. The difference between the ${\delta}^{13}C$ values of SOM and pelagic organic matter was also within 1%, suggesting that the SOM originated from the pelagic system, which is an important factor controlling the macrobenthic community.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.3
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• pp.256-260
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• 1999

Minimun-detectable levels to 28 antibacterial agents used for the prevention and the treatment of fish diseases were determined to establish optimal detective method of bioassay in fish by the EEC 4-plate method, the modified method of EEC 4-plate and the standard method of analysis in food safety regulation. The test organisms used in the methods of bioassay were as follows: Bacillus subtilis BGA (B. subtilis) and Micrococcus luteus ATCC 9341 (M. luteus) in the EEC 4-plate method, B. subtilis, M. luteus and Bacillus cereus var. mycoides ATCC 11778 (B. cereus) in the modified of EEC 4-plate, and B. subtilis, M. luteus, B. cereus and Bacillus stearothermophilis var. calidolactis C-953 (B. stearothermophilis) in the standard method. The standard method showed predominant sensitivity in the detection of penicillins (PCs), and was also highly sensitive to aminoglycosides (AGs). The sensitivity of standard method in the detection of tetracyclines (TCs), marrolides (MLs), nitrofuran derivatives(NFs) and quinolones (QNs) was very low, and against sulfonamides (SAs), however, was extremely low. The modified method of EEC 4-plate showed very high sensitivity to TCs. Both the EEC 4-plate and the modified method of EEC 4-plate showed competitively high sensitivity in the detection of PCs, MLs, NFs, QNs and SAs. All the methods studied in the experiment showed very low sensitivity against chloramphenicol (CMs). Consequently, the modified method of EEC 4-Plate was the best bioassay method with a wide range of sensitivity for the optimal detection of the residual antibacterial agents in fish.

• Microbiology and Biotechnology Letters
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• v.36 no.2
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• pp.87-95
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• 2008

Probiotics are live organisms that when administered in adequate amounts confer a health benefit on hosts. The administration of direct-fed microbials (DFM) such as lactobacilli and bacillus, may be a more direct approach to beneficially alter gastrointestinal microflora than altering dietary ingredients or supplementing with growth-promoting levels of antibiotics. It is apparent that microbes have an important influence on immune development and resistance to infection; that microbes are not static colonizers of our bodies, but are dynamic, symbiotic coresidents. And it can improve the surrounding environments; decrease the malodor caused by degrade the excrement. Recently, new paradigm such as environment protection and safe food have been settled. In domestic farm house, there is a great demand for probiotics as a substitute of antibiotics for the improvement of environmental quality and the production of a competitive goods. Probiotics circulated in a country have three categories: an animal medicine permitted by national veterinary research quarantine service (NVRQS), a support feed registered in city or country house, and not-registered goods. However, lots of unqualified goods were produced and circulated. And thus, it is in urgent need of evaluating the present situation and effect of probiotics. This study was conducted to evaluate the system of a probiotics as a livestock-environment improving agents for the alternation of antibiotics and quality control of it.

• Microbiology and Biotechnology Letters
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• v.45 no.3
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• pp.226-235
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• 2017

Carotenoids such as phytoene, lycopene, and ${\beta}-carotene$ are used as food colorants, animal feed supplements, and for human nutrition and cosmetic purposes. Previously, we reported the isolation of a novel marine bacterium, Kocuria gwangalliensis, which produces a pink-orange pigment. Phytoene desaturase (CrtI), encoded by the gene crtI, catalyzes lycopene formation from phytoene and is an essential enzyme in the early steps of carotenoid biosynthesis. CrtI is one of the key enzymes regulating carotenoid biosynthesis and has been implicated as a rate-limiting enzyme of the pathway in various carotenoid synthesizing organisms. Here, we report the cloning of the crtI gene responsible for lycopene biosynthesis from K. gwangalliensis. The gene consisted of 1,584 bases encoding 527 amino acid residues. The nucleotide sequence of the crtI gene was compared with that of other species, including Kocuria rhizophila and Myxococcus xanthus, and was found to be well conserved during evolution. An expression plasmid containing the crtI gene was constructed (pCcrt1), and Escherichia coli cells were transformed with this plasmid to produce a recombinant protein of approximately 57 kDa, corresponding to the molecular weight of phytoene desaturase. Lycopene biosynthesis was confirmed when the plasmid pCcrtI was co-transformed into E. coli containing the plasmid pRScrtEB carrying the crtE and crtB genes required for lycopene biosynthesis. The results from this study will provide valuable information on the primary structure of K. gwangalliensis CrtI at the molecular level.

• Journal of Wetlands Research
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• v.19 no.4
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• pp.409-422
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• 2017

In order to understand the environmental factors affecting the waterbird community in winter, we divided the watershed into 6 sections and investigated the size of the stream, the water quality, benthic invertebrates, and fish including birds. The influences on the community of water birds in Wonju stream were river structure, temperature and water quality. Among the structure of the river, the factors affecting the waterbird community in winter were the width of the stream and the width of the flowing water. The wider the width, the more species and the number of the winter waterbirds were. As the width of river water was wide, the smaller the width and width of the waterway, the fewer the population. The temperature changes of the wintering season affected the community of water birds because it led to freezing of rivers. The sections that were not frozen showed a significant increase and decrease in the temperature drop and rise. The values of pH, water temperature, COD, BOD, TN, TP and total E. coli increased with the increase of the downstream waterbirds population in the water quality survey. The water quality of S6 was affected by the downstream sewage treatment plant The number of wintering waterbirds was also highest. The effluent from the sewage treatment plant seems to have a considerable influence on the water quality. The increase of several items such as TN promotes the nutrition of the river, which leads to the accumulation of organic matter and the proliferation of aquatic organisms. This may be the cause of the increase in winter waterbirds as a food source. The benthic macroinvertebrate communities and fish communities did not show any correlation with the wintering water-birds communities.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.2
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• pp.174-190
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• 1996

Damage assessment based on in situ surveys for oil spills in marine environment is limited by fundamental difficulties as well as tremendous expenses. Except for intertidal zones, the damage is not preserved well. Also such surveys are usually confined to adult organisms. To overcome these limitations a computer model, NRDAM/CME, was developed in the case of USA (Reed et al., 1989), where an acute toxicity data base was used to assess indirect damages through food webs and loss due to recruitment as well as adult losses. In the present study damage assessment of natural biological resources for hypothetical oil spills is attempted using a computer model for hypothetical spills of Bunker C and heavy crude oil. In the model, the logical structure of NRDAM/CME was adopted, and biomass and productivity database were compiled for the Korean waters. The results showed that the damage increased in a nonlinear fashion as the spill amount increased. The magnitude of the damage depended upon the chemical properties of oil viscosity and solubility in particular, which implies that usage of oil dispersant might increase the damage by dispersing oil. The results also indicate that long term damage due to recruitment loss could be greater than short term damage.

• The Korean Journal of Malacology
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• v.19 no.1
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• pp.1-8
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• 2003

The present study was performed to describe the influence of water temperature and salinity on the filtration rates of the short-necked clam, Ruditapes philippinarum. The clams were collected at tidal flat near Yeosu city, Cheollanamdo, Korea, from July 2001 to August 2001. Diatoms, Phaeodactylum tricornutum (KMCC B-128), were indoor-cultured by f/2 medium, and were used to measure the filtration rate of the clams. Filtration rates of the clams were measured by indirect method. Cell concentrations of food organisms were determined by direct counting cells using the hemacytometer under the light microscope. The filtration rate of the clams increased with temperatures up to the optimum temperature, circa 25$^{\circ}C$. Above this optimum temperature, the filtration rate decreased drastically. Also the filtration rate of the clams increased with salinity up to 35 psu. The maximal filtration rates of the clams were recorded at 20-25$^{\circ}C$, similar to be known as the optimal temperature for their growth, and 25-35 psu, respectively. The minimal filtration rates of the clams were recorded at 5$^{\circ}C$ and 15 psu. At the similar temperature and salinity, the filtration rate of the younger clams was higher than that of the older ones. Thermal coefficient, Q$_{10}$ values at low temperature range were much higher than those at high temperature range. These results indicate the short-necked clam is more sensitive in cold water. As they grow up, they become more stronger against their ambient environmental changes, such as thermal-shock, salinity changes.

• ALGAE
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• v.32 no.4
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• pp.285-308
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• 2017

Cochlodinium polykrikoides red tides have caused great economic losses in the aquaculture industry in many countries. To investigate the roles of metazooplankton in red tide dynamics of C. polykrikoides in the South Sea of Korea, the abundance of metazooplankton was measured at 60 stations over 1- or 2-week intervals from May to November 2014. In addition, the grazing impacts of dominant metazooplankton on red tide species and their potential heterotrophic protistan grazers were estimated by combining field data on the abundance of red tide species, heterotrophic protist grazers, and dominant metazooplankton with data obtained from the literature concerning ingestion rates of the grazers on red tide species and heterotrophic protists. The mean abundance of total metazooplankton at each sampling time during the study was 297-1,119 individuals $m^{-3}$. The abundance of total metazooplankton was significantly positively correlated with that of phototrophic dinoflagellates (p < 0.01), but it was not significantly correlated with water temperature, salinity, and the abundance of diatoms, euglenophytes, cryptophytes, heterotrophic dinoflagellates, tintinnid ciliates, and naked ciliates (p > 0.1). Thus, dinoflagellate red tides may support high abundance of total metazooplankton. Copepods dominated metazooplankton assemblages at all sampling times except from Jul 11 to Aug 6 when cladocerans and hydrozoans dominated. The calculated maximum grazing coefficients attributable to calanoid copepods on C. polykrikoides and Prorocentrum spp. were 0.018 and $0.029d^{-1}$, respectively. Therefore, calanoid copepods may not control populations of C. polykrikoides or Prorocentrum spp. Furthermore, the maximum grazing coefficients attributable to calanoid copepods on the heterotrophic dinoflagellates Polykrikos spp. and Gyrodinium spp., which were grazers on C. polykrikoides and Prorocentrum spp., respectively, were 0.008 and $0.047d^{-1}$, respectively. Therefore, calanoid copepods may not reduce grazing impact by these heterotrophic dinoflagellate grazers on populations of the red tide dinoflagellates.