• Journal of Soil and Groundwater Environment
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• v.27 no.6
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• pp.11-21
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• 2022

Sorption of phenanthrene (PHE) and pyrene (PYR) in several sorbents, i.e., natural soil, BionSoil^®, Pahokee peat, vermicompost and Devonian Ohio Shale and a surfactant (hexadecyltrimethyl ammonium chloride)-modified montmorillonite (HDTMA-M) were investigated. Pyrene exhibited higher sorption tendency than phenanthrene, as predicted by its higher octanol to water partition coefficient (K_ow). Several sorption models: linear, Freundlich, solubility-normalized Freundlich model, and Polanyi-Manes model (PMM) were used to analyze sorption isotherms. Linear isotherms were observed for natural soil, BionSoil^®, Pahokee peat, vermicompost, while nonlinear Freundlich isotherms fitted for Ohio shale and HDTMA-M. The relationship between sorption model parameters, organic carbon content (f_oc), and elemental C/N ratio was studied. In the binary competitive sorption of phenanthrene and pyrene in natural soil, competition between the solutes caused reduction in the sorption of each solute compared with that in the single-solute system. The ideal adsorbed solution theory (IAST) coupled with the single-solute Freundlich model was not successful in describing the binary competitive sorption equilibria. This was due to the inherent nature of linear sorption of phenanthrene and pyrene in natural soil. The result indicates that the applicability of IAST for the prediction of binary competitive sorption is limited when the sorption isotherms are inherently linear.

• Journal of Microbiology and Biotechnology
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• v.25 no.7
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• pp.988-998
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• 2015

The filamentous fungus Aspergillus oryzae is a well-known expression host used to express homologous and heterologous proteins in a number of industrial applications. To facilitate higher yields of proteins of interest, we constructed the pAsOP vector to express heterologous proteins in A. oryzae. pAsOP carries a selectable marker, pyrG, derived from Aspergillus nidulans, and a strong promoter and a terminator of the amyB gene derived from A. oryzae. pAsOP transformed A. oryzae efficiently via the PEG-CaCl₂-mediated transformation method. As proof of concept, green fluorescent protein (GFP) was successfully expressed in A. oryzae transformed by pAsOP-GFP. Additionally, we identified a novel fungal α-amylase (PcAmy) gene from Penicillium sp. and cloned the gene into the vector. After transformation by pAsOPPcAmy, the α-amylase PcAmy from Penicillium sp. was successfully expressed in a heterologous host system for the first time. The α-amylase activity in the A. oryzae transformant was increased by 62.3% compared with the untransformed A. oryzae control. The PcAmy protein produced in the system had an optimum pH of 5.0 and optimum temperature of 30^oC. As a cold-adapted enzyme, PcAmy shows potential value in industrial applications because of its high catalytic activity at low temperature. Furthermore, the expression vector reported in this study provides promising utility for further scientific research and biotechnological applications.

• Journal of Microbiology and Biotechnology
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• v.11 no.5
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• pp.812-818
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• 2001

Alginate and alginate-derived oligomannuronate enhanced penicillin production in shake flask and fermentor cultures of Penicillium chrysogenum Wis 54-1255 (containing a single copy of the penicillin gene cluster) and in the high producter strain P. chrysogenum AS-P-99 (containing multiple copies of the penicillin gene cluster). Alginate was not used as a single carbon source by P. chryogenum. The stimulatory effect on penicillin production was observed in a defined medium and, to a lower extent, in a complex production medium containing corn steep liquor. Alginate-supplemented cells showed higher transcript levels of the three penicillin biosynthetic genes, pcbAB, pcbC, and penDE, than cells grown in the absence of alginate. The promoters of the pcbAB, pcbC, and penDE genes were coupled to the reporter lacZ gene and introduced as monocopy constructions in P. chrysogenum Wis 54-1225 npe10 by targeted integration in the pyrG locus; the reporter ${\beta}$-galactosidase activity expressed from the three promoters was stimulated by alginate added to the culture medium of the transformants. These results indicate that the stimulation of penicillin production by alginate was derived from an increase in the transcriptional activity of the penicillin biosynthesis genes. The induction by alginate of the transcription of the three penicillin biosynthetic genes is good example of the coordinated induction of secondary metabolism genes by elicitors of plant (or microbial) origin.

• Proceedings of the Microbiological Society of Korea Conference
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• 2001.11a
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• pp.157-163
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• 2001

Mycolic acid-containing gram-positive bacteria, so called nocardioform actinomycetes, have become a great interest to environmental microbiologists due to their metabolic versatility, multidegradative capacity and potential for bioremediation of priority pollutants. For example, Rhodococcus rhodochrous N75 was able to metabolize 4-methy1catechol via a modified $\beta$-ketoadipate pathway whereby 4-methylmuconolactone methyl isomerase catalyzes the conversion of 4-methylmuconolactone to 3-methylmuconolactone in order to circumvent the accumulation of the 'dead-end' metabolite, 4-methylmuconolactone. R. rhodochrous N75 has also shown the ability to transform a range of alkyl-substituted catechols to the corresponding muconolactones. A novel 3-methylmuconolactone-CoAsynthetase was found to be involved in the degradation of 3-methylmuconolactone, which is not mediated in a manner analogous to the classical $\beta$-ketoadipate pathway but activated by the addition of CoA prior to hydrolysis of lactone ring, suggesting that the degradative pathway for methylaromatic compounds by gram-positive bacteria diverges from that of proteobacteria. Mycobacterium sp. Strain PYR-l isolated from oil-contaminated soil was capable of mineralizing various polyaromatic hydrocarbons (PAHs), such as naphthalene, phenanthrene, pyrene, fluoranthrene, 1-nitropyrene, and 6-nitrochrysene. The pathways for degradation of PAHs by this organism have been elucidated through the isolation and characterization of chemical intermediates. 2-D gel electrophoresis of PAH-induced proteins enabled the cloning of the dioxygenase system containing a dehydrogenase, the dioxygenase small ($\beta$)-subunit, and the dioxygenase large ($\alpha$)-subunit. Phylogenetic analysis showed that the large a subunit did not cluster with most of the known sequences except for three newly described a subunits of dioxygenases from Rhodococcus spp. and Nocardioides spp. 2-D gel analysis also showed that catalase-peroxidase, which was induced with pyrene, plays a role in the PAH metabolism. The survival and performance of these bacteria raised the possibility that they can be excellent candidates for bioremediation purposes.

• Korean journal of food and cookery science
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• v.10 no.3
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• pp.301-307
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• 1994

Benzo(a)pyrene [B(a)P], one of the polycyclic aromatic hydrocarbons(PAHs) is know as a potent careinogen. As consumption of lipid food and fried food increases recently, the toxic effect of overheated lipid foods has report increasingly. This study was undertaken to measure the contents of B(a)P and other PAHs of fried chicken and frying oil. At the same time, the rancidities of frying oil and descarded oil were determined. B(a)P and other PAHs contents of all samples were measured by HPLC/UV method. The results obtained were as follows; B(a)P contents of fresh chicken and soybean oil were 0.073 and 0.146 ${\mu}$g/kg, respectively. During fried chicken at 180${\pm}$5$^{\circ}C$ and 200${\pm}$5$^{\circ}C$, B(a)P and PAHs contens decreased or increased In 20 times fried chicken, PAHs contents except Pyr and DMBA increased at 180${\pm}$5$^{\circ}C$ and PAHs contents except DMBA, DBA, and 3-MC increased at 200${\pm}$5$^{\circ}C$. In frying oils fried chicken, PAHs contents increased in 10 and 15 times frying oil at 180${\pm}$5$^{\circ}C$ and grew more increase and more increase during frying at 200${\pm}$5$^{\circ}C$. The changing pattern of PAHs contents of frying oils was remotely related that of rancidities of frying oils. In fried chicken, frying oils, and discarded oils obtained from fried chicken house at market, PAHs contents of fried chickens were lower than those of fried chicken cooked at laboratory but those of frying oils and discarded oils were higher than those of frying oils cooked at laboratory.

• Journal of Microbiology and Biotechnology
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• v.25 no.2
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• pp.196-205
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• 2015

A Sulfolobus-E. coli shuttle vector for an efficient expression of the target gene in S. acidocaldarius strain was constructed. The plasmid-based vector pSM21 and its derivative pSM21N were generated based on the pUC18 and Sulfolobus cryptic plasmid pRN1. They carried the S. solfataricus P2 pyrEF gene for the selection marker, a multiple cloning site (MCS) with C-terminal histidine tag, and a constitutive promoter of the S. acidocaldarius gdhA gene for strong expression of the target gene, as well as the pBR322 origin and ampicillin-resistant gene for E. coli propagation. The advantage of pSM21 over other Sulfolobus shuttle vectors is that it contains a MCS and a histidine tag for the simple and easy cloning of a target gene as well as one-step purification by histidine affinity chromatography. For successful expression of the foreign genes, two genes from archaeal origins (PH0193 and Ta0298) were cloned into pSM21N and the functional expression was examined by enzyme activity assay. The recombinant PH0193 was successfully expressed under the control of the gdhA promoter and purified from the cultures by His-tag affinity chromatography. The yield was approximately 1 mg of protein per liter of cultures. The enzyme activity measurements of PH0913 and Ta0298 revealed that both proteins were expressed as an active form in S. acidocaldarius. These results indicate that the pSM21N shuttle vector can be used for the functional expression of foreign archaeal genes that form insoluble aggregates in the E. coli system.

• Journal of fish pathology
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• v.32 no.2
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• pp.69-80
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• 2019

We performed MLSA (multilocus sequence analysis) and phenotypic characterization of Aliivibrio species isolated from walleye pollock (Gadus chalcogrammus) maintained in 3 different facilities of Gangwon Province, the east coast of Korea. Of 38 Aliivibrio species identified by 16S rDNA sequences, 12 strains were randomly selected and MLSA was conducted with 5 house-keeping genes (gapA, gyrB, pyrH, recA and rpoA) and 16S rDNA gene. Phylogenetic analysis and homology of the concatenated sequences (4,580 bp) with other Vibrionaceae genera revealed that 4 strains (GNGc16.1, YYGc16.1, YYGc16.2, GSGc18.1) were identified as Aliivibrio logei and one strain (GSGc16.1) as A. wodanis. One strain (GSGc17.1) was tentatively identified as A. logei, but needs further analysis because it did not belong to the same clade with A. logei type strain. 6 strains (GSGc17.2, GNGc16.2, GSGc16.2, GSGc17.3, GSGc18.2, GSGc17.4) need further investigation as potential novel species. Either phenotypic characterization or 16S rDNA sequence alone did not provide enough information for identification of Aliivibrio strains at the species level. A. logei and A. wodanis are generally known as non-pathogenic bacteria, but also known as opportunistic or secondary pathogens of cold water fishes. Cares should be taken to prevent potential outbreaks due to these bacteria, although there was no outbreaks during the sampling period.

• Korean journal of food and cookery science
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• v.9 no.4
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• pp.323-328
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• 1993

Benzo(a)pyrene [B(a)P], one of the polycyclic aromatic hydrocarbons (PAHs) is known as a potent carcinogen. As lipid consumption increases recently, the toxic effect of overheated lipid foods and fats ＆ oils were reported increasingly. In this study, the contents of B(a)p, other PAHs and rancidities of soybean oil were determined, and then the proper heating temperature, time and frequency were recommended, The work was carried out using soybean oil heated at $180\pm$5^{\circ}C$$, $200\pm$5^{\circ}C$$, and $300\pm$5^{\circ}C$$,for 50 hours. Acid Value(AV) and Conjugated Diene Value of samples were determined. The contents of B(a)P and other PAHs contents of all samples were masured by HPLC/UV method. The results obtained were as follows; Each content of PAHs in the fresh soybean oil was: Pyr 1.093, B(a)A 0.986, Ch 1.147, DMBA 1.082, B(e)P 0.664, Per 1.135, B(a)P 0.146, DBA 1.053, 3-MC 0.05 rg/kg. When the soybean oil was heated at $180\pm$5^{\circ}C$$, for 10, 20, 30, and 50 hours, B(a)P conterlts in heated soybean oils were 0.391, 0.692, 0.451, and 0.372 $\mu\textrm{g}$/kg respectively. Acid value of them were 0.26, 0.26, 0.29, and 0.33, and conjugated diene value was 0.67, 0.76, 0.99, and 1.04, respectively. When the soybean oil was heated at $200\pm$5^{\circ}C$$,and $300\pm$5^{\circ}C$$,for 10, 20, 30, and 50 hours, B(a)P coiltents in soybean oil heated at $200\pm$5^{\circ}C$$,were 0.844, 0.512, 0.479 and 0.247 Ig/kg respectively, Acid value 0.22, 0.21, 0.23 and 0.51 and CDNV 0.39, 0.49, 3.27, and 3.89. B(a)P contents in soybean oil heated at $300\pm$5^{\circ}C$$,were 0.466, 0.706, 0.607 and 0.247$\mu\textrm{g}$/kg respectively, Acid value 0.47, 1.57, 3.90, and 6.42 and CDNV 0.65, 2.15, 3.00, and 3.88.

• Journal of Life Science
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• v.22 no.1
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• pp.98-109
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• 2012

The properties of lactate dehydrogenase (EC 1.1.1.27, LDH) and expression of monocarboxylate transporters (MCTs) 1, 2, and 4 were studied in tissues from Micropterus salmoides. Native-PAGE revealed that the LDH $A_4$ isozyme was predominantly located in skeletal muscle. The LDH $A_4$, $A_2B_2$, and $B_4$ isozymes were detected in heart, liver, eye, and brain tissues, while eye-specific $C_4$ isozyme was detected in eye tissue. In September, strong LDH $B_4$ isozyme activity was detected in heart tissue. High $A_4$ isozyme activity was noted in all other tissues except heart tissue. However, in November, strong $A_4$ isozyme activity was detected in heart tissue. The LDH/CS (Citrate synthase, EC 4.1.3.7) ratio in skeletal muscle and heart tissues indicated that anaerobic metabolism was high in those tissues. Native-PAGE after immunoprecipitation showed that eye-specific $C_4$ isozyme was more similar to the $A_4$ than the $B_4$ isozyme. The LDH $A_4$ isozyme was purified by affinity chromatography. The molecular weight of subunit A was 37,200. The LDH activity in tissues was consistently 11.05~28.32% due to inhibition by 10 mM pyruvate. The $K_m^{PYR}$ of LDH in eye tissue was very low. The optimum pH for LDH in tissues was pH 7.5~8.0. The LDH $A_4$ isozyme was detected in mitochondria of skeletal muscle, whereas the $B_4$ and $A_2B_2$ isozymes were detected in heart tissue mitochondria. Western blot analysis indicated that MCTs 1, 2, and 4 were located in the plasma membrane and mitochondria of skeletal muscle and heart tissues. The sizes of MCTs 1, 2, and 4 in skeletal muscle were 60, 54~38, and 63 kDa, while those in heart tissue were 57, 54~38, and 55.5 kDa, respectively. In conclusion, M. salmoides appears to use anaerobic metabolism predominantly when adapted to a hypoxic environment. In highly activated skeletal muscle and heart tissue, energy production is controlled by inward and outward flows of pyruvate and lactate through MCTs 1, 2, and 4 in the plasma membrane and mitochondria, with effective adjustment by LDH isozymes.

• Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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• v.2 no.1
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• pp.57-68
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• 1998

Polycyclic aromatic hydrocarbons (PAMs) are ubiquitous contaminants in marine environments. PAMs enter estuarine and nearshore marine environment via several routes such as combustion of fossil fuels, domestic and industrial effluents and oil spills. PAHs have been the focus of numerous studies in the world because they are potentially carcinogenic, mutagenic, and teratogenic to aquatic organisms and humans from consuming contaminated food. However, one can hardly find any available data on PAM content in marine organisms in Korea. The present study was carried out in order to determine PAM content in oysters from the intertidal and subtidal zones of Chinhae Bay, which is located in near urban communities and an industrial complex, and the bay is considered to be a major repositories of PAHs. 16 PAHs were analyzed by High Performance Liquid Chromatography (HPLC) with uv/vis and fluorescence detectors in oysters: they are naphthalene (NPTHL), acenaphthylene (ANCPL), acenaphthylene (ACNPN), fluorene (FLURN), phenanthrene (PHEN), anthracene (ANTHR), fluoranthene (FLRTH), pyrene (PYR), benzo(a)anthracene (BaA), chrysene (CHRY), benzo(b)fluoranthene (BbF), benzo(k)fluoranthene (BkF), benzo(a)pyrene (BaP), dibenz(a,h)anthracene(DahA), benzo(g,h,i)peryne (BghiP) and indeno(1,2,3,-cd)pyrene (I123cdP). The PAM contents in oysters from the intertidal and subtidal zones of Chinhae Bay ranged from < 0.1 to 992.0 ug/kg (mean $69.8\pm9.8$ ug/kg).