• KSBB Journal
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• v.27 no.6
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• pp.352-360
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• 2012

Bacillus clausii I-52 which produced SDS- and $H_2O_2$-tolerant extracellular alkaline protease (BCAP) was isolated from heavily polluted tidal mud flat of West Sea in Incheon, Korea and stable strain (transformant C5) of B. clausii I-52 harboring another copy of BCAP gene in the chromosome was developed using the chromosome integration vector, pHPS9-fuBCAP. When investigated the production of BCAP using B. clausii transformant C5 through pilot-scale submerged fermentation (500 L) at $37^{\circ}C$ for 30 h with an aeration rate of 1 vvm and agitation rate of 250 rpm, protease yield of approximately 105,700 U/mL was achieved using an optimized medium (soybean meal 2%, wheat flour 1%, sodium citrate 0.5%, $K_2HPO_4$ 0.4%, $Na_2HPO_4$ 0.1%, NaCl 0.4%, $MgSO_4{\cdot}7H_2O$ 0.01%, $FeSO_4{\cdot}7H_2O$ 0.05%, liquid maltose 2.5%, $Na_2CO_3$ 0.6%). The enzyme stability of BCAP was increased by addition of polyols (10%, v/v) and also, the stabilities of BCAP towards not only the thermal-induced inactivation at $50^{\circ}C$ but also the SDS and $H_2O_2$-induced inactivation at $50^{\circ}C$ were enhanced. Among the polyols examined, the best result was obtained with propylene glycol (10%, v/v). The BCAP supplemented with propylene glycol exhibited extreme stability against not only the detergent components such as ${\alpha}$-orephin sulfonate (AOS) and zeolite but also the commercial detergent preparations. The granulized enzyme of BCAP was prepared with approximately 1,310,000 U/g of granule. Wash performance analysis using EMPA test fabrics revealed that BCAP granule exhibited high efficiency for removal of protein stains in the presence of anionic surfactants as well as bleaching agents. When compared to Savinase 6T$^{(R)}$ and Everlase 6T$^{(R)}$ manufactured by Novozymes, BCAP under this study probably showed similar or higher efficiency for the removal of protein stains. These results suggest that the alkaline protease produced from B. clausii transformant C5 showing high stability against detergents and high wash performance has significant potential and a promising candidate for use as a detergent additive.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.5 no.1
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• pp.1-9
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• 2000

Vertical temperature distributions in muddy intertidal sediments near the Jebu Island on the west coast of Korea were obtained during the period of ebb tide which occurred in day time. The observations of mud temperature were made with thermistor embedded probe at 2cm interval for 18cm-layer of sediment for five different months of the year. Temporal changes in the vertical profile of the sediment temperature are strongly depend on the air temperature, the previous time of flood tide and the time of ebb tide. Heat exchanges in the surface layer (0-2 cm) in terms of magnitude and direction are greater than and opposite to those in the deeper sediment layer (8-12 cm), respectively and do not show any significant seasonal variations. In general, the surface layer gains heat while the deeper layer loses the heat. By using the 1-D diffusion equation temporal vertical profiles of the sediment temperature were obtained and were compared with the observed ones. The results show that in the sediment layer below 4 cm-depth the heat transport is predominantly by molecular diffusion. The average magnitude of heat flux into the sediment layer (0-18 cm) during the ebb tide when the mudflats were exposed in the middle of the day were between 4.1 and $28.9\;W/m^2$.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.3 no.2
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• pp.94-102
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• 1998

Temporal changes, density, calcimass, mode of occurrence, size-frequency histogram and survivorship curves were studied for the molluscan death assemblages on a macrotidal flat, Inchon, Korea. The living and death assemblages were compared on the basis of the taxonomic compositions and their numerical abundances. A total of 28 species (16 taxa in gastropods and 12 in bivalves) were identified. Most of the dead shells were the species inhabiting that intertidal mud flat. Species diversity was higher in gastropods than in bivalves. Seasonal variation of the calcimass was influenced by the mactrid bivalve, Mactra veneriformis. Shell-boring naticid gastropods preferred selecting the umbo areas of various valves. Size-frequency distribution and size-specific survivorship curves were analyzed for the dominant species. Survivorship curve of the tellinid bivalve, Morella rutila was convex-up in shape, which is congruent with the expected equilibrium condition but indicates higher mortality in winter. Whereas the survivorship curves of Mactra veneriformis and Reticunassa festiva were semi convex-up with dual modes, suggesting a disequilibrium due to the changes in recruitment and seasonal mortality. This study suggests that the dead shell assemblages may be useful in getting population information like live molluscan assemblages, if solved for several problems related to taphonomic processes.

• Journal of Life Science
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• v.25 no.2
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• pp.189-196
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• 2015

Bacterial diversity was studied in the rhizosphere of Suaeda japonica Makino, which is native to Suncheon Bay in South Korea. Soil samples from several sites were diluted serially, and pure isolation was performed by subculture using marine agar and tryptic soy agar media. Genomic DNA was extracted from 29 pure, isolated bacterial strains, after which their 16S rDNA sequences were amplified and analyzed. Phylogenetic analysis was performed to confirm their genetic relationship. The 29 bacterial strains were classified into five groups: phylum Firmicutes (44.8%), Gamma proteobacteria group (27.6%), Alpha proteobacteria group (10.3%), phylum Bacteriodetes (10.3%), and phylum Actinobacteria (6.8%). The most widely distributed genera were Bacillus (phylum Firmicutes), and Marinobacterium, Halomonas, and Vibrio (Gamma proteobacteria group). To confirm the bacterial diversity in rhizospheres of S. japonica, the diversity index was used at the genus level. The results show that bacterial diversity differed at each of the sampling sites. These 29 bacterial strains are thought to play a major role in material cycling at Suncheon Bay, in overcoming the sea/mud flat-specific environmental stress. Furthermore, some strains are assumed to be involved in a positive interaction with the halophyte S. japonica, as rhizospheric flora, with induction of growth promotion and plant defense mechanism.

• Microbiology and Biotechnology Letters
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• v.42 no.4
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• pp.324-330
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• 2014

A marine bacterial strain producing agar-degrading enzymes was isolated from a mud flat in Jeboo-do (Korea) using a selective artificial sea water (ASW) agar plate containing agar as the sole carbon source. The isolate, designated as SH-1, was gram-negative, aerobic, and motile with single polar flagellum. 16S rRNA gene sequence similarity analysis showed the isolate SH-1 had the highest homology (96.5%) to marine bacterium Neiella marina J221. Cells could grow at $28-37^{\circ}C$ but not at $42^{\circ}C$, and the agarase activity of the cell culture supernatant was higher when grown at $28^{\circ}C$ than when grown at $37^{\circ}C$. Cells could grow when concentrations of 1-5% (w/v) NaCl were added to the growth media with the best growth observed at 3% NaCl, and the agardegrading enzyme activity of the cell culture supernatant was best when grown at 3% NaCl-containing growth media under the conditions we examined. The crude enzyme prepared from 48-h culture broth of strain SH-1 exhibited an optimum pH and temperature for agar-degrading activity at 7.0 and $40^{\circ}C$, respectively. Zymogram analysis of the crude supernatant and cell extract showed that strain SH-1 produced at least 3 agar-degrading enzymes with molecular weights of 15, 35, and 52 KD. Thinlayer chromatography (TLC) analysis also suggested that HS-1 produces ${\beta}$-agarase to degrade agarose to neoagarooligosaccharides.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.8 no.4
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• pp.380-391
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• 2003

Surface sediments were collected from Gwangyang and Yeosu Bays to evaluate their sedimentological characteristics and geochemical aspects of both the benthic environment and heavy metal pollution. The grain size distribution includes both sandy and muddy sediments. Sand-rich sediments occur mainly near the POSCO and the channel between Namhedo and Yeosu Bando, while elsewhere mud-dominated sediments are present. TOC content ranges from 0.2 to 2.1 % and C/N ratios indicate that the range arises from the mix of organic matter. The C/S ratios of this organic matter show that parts of the study area are anoxic or have sub-anoxic bottom conditions. The hydrogen sulfide content of the sediment has a range of 0.7 to 301 ppm, with a high content occurring inshore of Myodo Island, where it indicates a polluted environment. The enrichment factor (Ef) and index of accumulation rate (Igeo) of ten heavy metals (Co, Ni, Cu, Cd, Pb, Li, Zn, V, Cr, Ba) show that parts of the study area contain from one to seven times more Pb and Ba, and from 0.8 to 3.5 times more of the other elements than the mean sediment value. The Igeo values of V and Cd show that different parts of the area can be classified as heavily polluted, heavily to moderately polluted, or more or less unpolluted. Those areas that have both high levels of enrichment and high accumulation rates of heavy metals contain predominantly fine sediments with a high organic matter and hydrogen sulfide content.

• Journal of agriculture & life science
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• v.45 no.6
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• pp.201-212
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• 2011

The alkaline protease gene was cloned from a halo-tolerant alkalophilic Bacillus clausii I-52 isolated from the heavily polluted tidal mud flat of West Sea in Inchon Korea, which produced a strong extracellular alkaline protease (BCAP). Based on the full genome sequence of Bacillus subtilis, PCR primers were designed to allow for the amplification and cloning of the intact pro-BCAP gene including promoter region. The full-length gene consists of 1,143 bp and encodes 381 amino acids, which includes 29 residues of a putative signal peptide and an additional 77-amino-acid propeptide at its N-terminus. The mature BCAP deduced from the nucleotide sequence consists of 275 amino acids with a N-terminal amino acid of Ala, and a relative molecular weight and pI value was 27698.7 Da and 6.3, respectively. The amino acid sequence shares the highest similarity (99%) to the nattokinase precursor from B. subtilis and subtilisin E precursor from B. subtilis BSn5. The substrate specificity indicated that the recombinant BCAP could hydrolyze efficiently the synthetic substrate, N-Suc-Ala-Ala-Pro-Phe-pNA,and did not hydrolyze the substrates with basic amino acids at the P1 site. The recombinant BCAP was strongly inhibited by typical serine protease inhibitor, PMSF, indicating that BCAP is a member of the serine proteases.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.28 no.4
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• pp.143-157
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• 2023

This study confirmed the characteristics of macrobenthic community due to anthropogenic environmental changes in the Boryeong Jugyo tidal flat, where the habitat of manila clam (Ruditapes philippinarum) and mud shrimp (Upogebia major) is separated. The total number of occurring species was 55 during the study period with an average habitat density of 338 ind./m² and a biomass of 212.2 gWWt/m². The number of occuring species increased from 27 species at the upper flat to 37 species at the lower flat, and the dominant species differed by tide levels (Upper: Leonnates persica, Middle: Heteromastus filiformis, Lower: R. philippinarum). The macro-benthic community sturctures of the top 10 species using cluster analysis and nMDS were divided into two groups, focusing on Manila clam culture farm of lower flats and middle flats with high habitat density, reflecting the influence of specific species. The sediment composition of the U. major habitat space fluctuated highly, but it was maintained annually, and the sorting coefficient was 2.1 𝜑, and the proportion of the same particle size was increased. In particular, because the middle flat has a dense anthropogenic impact, a dominant species, H. filiformis dominated and revealed a relationship with the density of burrow holes of U. major, which is considered to be a biological interaction between these two macrofauna in this tidal flat.

• Economic and Environmental Geology
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• v.30 no.4
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• pp.303-312
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• 1997

The massive, fractured and porous-type of glauconite, which is subdivided by surface morphology, occur in subtidal sand and semiconsolidated intertidal sand/mud from continental shelf of the southeastern Yellow Sea. This area is presumed to be a part of Holocene transgressive tidal systems tract. The glauconite, pellet-like grains with diameter of 0.1 to 1 mm, is scattered in surface sand sediments. Results of X-ray diffraction data of the minerals are monoclinic with $a=5.242{\AA}$, $b=9.059{\AA}$, $c=10.163{\AA}$, ${\beta}=100.5^{\circ}$, $V=474.53{\AA}^3$. Thermal treatments on the oriented glauconite increase the X-ray diffraction intensity near $10{\AA}$ (001), suggesting the presence of some expandable layers. Specific gravity of the glauconite is $2.60{\pm}0.45gm/cc$ on the basis of chemical composition and unit-cell dimensions. Based on $O_{10}(OH)_2$, chemical composition of glauconites, octahedral Fe content ranges from 1.19 to 2.06 atoms, corresponding octahedral AI is 0.18 to 0.76 atoms, which progressively substitute Fe for AI with increasing from porous to massive-type. The Mg content ranges from 0.35 to 0.54 atoms, and shows higher with increasing Al contents. A systematic increase of interlayer K from 0.34 to 0.71 is also observed with apparent increases from porous to massive-type, and related to a proportion of expandable layers. The clay preserved in glauconite, which is recognized as ordered/disordered (massive to fractured-type). The interstratified illite/smectite (porous-type), contains 7 to 27 % expandable layers. The glauconite seems to originate from post depositional authigenic growth in reducing environments promoted by the dissolution of clay minerals and biogenic debris.

• Applied Biological Chemistry
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• v.41 no.7
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• pp.533-538
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• 1998

In order to gain fundamental information on the utilization of lipids as energy source in mud-skipper, muscle lipids and their fatty acid composition were investigated with respect to life cycle-maturation (Aug), before-hibernation (Nov), and after-hibernation (Apr). Crude and neutral lipid were found to decrease from 1.2, 68.3% (Aug) via 0.7, 53.8% (Nov) to 0.4, 42.6% (Apr), respectively, whilst phospholipids and glycolipids increased from 29.5 and 2.2% (Aug) to 52.1 and 5.3% (Apr), respectively. In neutral lipids, TG contents gradually decreased from 53.8% (Aug) via 33.6% (Nov) to 23.1% (Apr), while FFA and sterol contents increased from 13.5 and 14.2% (Aug) to 22.3 and 24.5% (Apr), respectively. In phospholipids, PC content decreased from 61.2% (Aug) to 50.6% (Apr), while changes in PS and PE contents, as a whole, showed the opposite trends. In neutral lipids, the levels of some fatty acids such as 16:0, 16:1, 18:0, 18:1 and 20:5 $({\omega}3)$ were analyzed to be high, with the 20 : 5 being predominant, and the levels of saturated and monoene-acids gradually decreased, while polyene-acids increased in before and after hibernation. In before hibernation, 16:0, 16:1, 18:1 mainly decreased but 18:2, 18:3 $({\omega}3)$ and most of saturated and monoene acids such as 14:0, 14:1, 16:1, 18:0, 18:1 slightly decreased in after hibernation. From these findings, it was suggested that those fatty acids decreased during hibernation were used as a energy source, particularly 16:1 and 18:1 being most preferentially used. In phospholipids, the levels of 16:0, 18:0, 20:5 $({\omega}3)$, 22:5 $({\omega}3)$ and 22:6 $({\omega}3)$ were found to be high. Throughout the life cycle, the levels of monoene-acids in phospholipids stayed constant, whilst those of 18:2, 18:3 $({\omega}3)$ and saturated acids such as 16:0, 18:0 were found to be decreased gradually in before and after hibernation, whereas those for the high degree of polyene-acids such as 20:4 $({\omega}6)$, 20:5 $({\omega}3)$, 22:5 $({\omega}3)$, 22:6 $({\omega}3)$ increased, particularly 20:4 $({\omega}6)$, 20:5 $({\omega}3)$ being most increased.