• Microbiology and Biotechnology Letters
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• v.42 no.1
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• pp.18-24
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• 2014

This study was conducted to investigate optimum conditions for the production of cellulose-degrading crude enzymes by an isolated marine bacterium. A marine microorganism producing an extracellular cellulose-degrading enzyme was isolated from the red seaweed, Grateloupia elliptica Holmes. The isolated bacterium was identified as Cellulophaga lytica by 16S ribosomal RNA gene sequence analysis and physiological profiling and designated as Cellulophaga lytica PKA 1005. The optimum conditions for the growth of Cellulophaga lytica PKA 1005 were pH 7, 2% NaCl, and $30^{\circ}C$ with 36 h incubation time. To obtain the crude enzyme, the culture medium of the strain was centrifuged for 30 min at $12,000{\times}g$ and $4^{\circ}C$, and the supernatant was used as crude enzyme. The optimum conditions for the production of the cellulose-degrading crude enzyme were pH 8, $35^{\circ}C$, 8% carboxyl methyl cellulose, and 60 h reaction time.

• Microbiology and Biotechnology Letters
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• v.48 no.2
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• pp.215-222
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• 2020

The marine microorganism PX-1, which can hydrolyze xylan, was isolated from coastal sea water of Jeju Island, Korea. Based on the 16S rRNA gene sequence and chemotaxonomy analysis, PX-1 was identified as a species of the genus Aestuariibacter and named Aestuariibacter sp PX-1. From the culture broth of PX-1, an extracellular xylanase was purified to homogeneity through ammonium sulfate precipitation and subsequent adsorption chromatography using insoluble xylan. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration chromatography estimated the molecular weight of the purified putative xylanase (XylA) as approximately 64 kDa. XylA showed xylanase activity toward beechwood xylan, with a maximum enzymatic activity at pH 6.0 and 45℃. Through thin-layer chromatographic analysis of the xylan hydrolysate produced by XylA, it was confirmed that XylA is an endo-type xylanase that decomposes xylan into xylose and xyloligosaccharides of various lengths. The K_m and V_max values of XylA for beechwood xylan were 27.78 mM and 78.13 μM/min, respectively.

• Journal of Life Science
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• v.23 no.4
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• pp.542-553
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• 2013

The aim of this work was to establish the optimal conditions for the production of cellobiase by a marine bacterium, Cellulophaga lytica LBH-14, using response-surface methodology (RSM). The optimal conditions of rice bran, ammonium chloride, and the initial pH of the medium for cell growth were 100.0 g/l, 5.00 g/l, and 7.0, respectively, whereas those for the production of cellobiase were 91.1 g/l, 9.02 g/l, and 6.6, respectively. The optimal concentrations of $K_2HPO_4$, NaCl, $MgSO_4{\cdot}_{7H2}O$, and $(NH_4)_2SO_4$ for cell growth were 6.25, 0.62, 0.28, and 0.42 g/l, respectively, whereas those for the production of cellobiase were 4.46, 0.36, 0.27, and 0.73 g/l, respectively. The optimal temperatures for cell growth and for the production of cellobiase by C. lytica LBH-14 were 35 and $25^{\circ}C$, respectively. The maximal production of cellobiase in a 100 L bioreactor under optimized conditions in this study was 92.3 U/ml, which was 5.4 times higher than that before optimization. In this study, rice bran and ammonium chloride were developed as carbon and nitrogen sources for the production of cellobiase by C. lytica LBH-14. The time for the production of cellobiase by the marine bacterium with submerged fermentations was reduced from 7 to 3 days, which resulted in enhanced productivity of cellobiase and a decrease in its production cost. This study found that the optimal conditions for the production of cellobiase were different from those of CMCase by C. lytica LBH-14.

• Journal of Life Science
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• v.23 no.6
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• pp.757-769
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• 2013

A microorganism producing carboxymethylcellulase (CMCase) was isolated from seawater, identified as Bacillus velezensis by analyses of 16S rDNA and partial sequences of the gyrA, and designated as B. velezensis A-68. The optimal conditions for production of CMCase by B. velezensis A-68 were established using response surface methodology (RSM). The optimal concentrations of rice hulls and yeast extract, and initial pH of the medium for cell growth were 60.2 g/l, 7.38 g/l, and 7.18, respectively, whereas those for production of CMCase were 50.0 g/l, 5.00 g/l, and 7.30. The analysis of variance (ANOVA) implied that the most significant factor for cell growth as well as production of CMCase was yeast extract. The optimal concentrations of $K_2HPO_4$, NaCl, $MgSO_4{\cdot}7H_2O$, and $(NH_4)_2SO_4$ in the medium for cell growth were 7.50, 1.00, 0.10, and 0.80 g/l, respectively, which were the same as those for production of CMCase. The optimal temperatures for cell growth and production of CMCase were 30 and $35^{\circ}C$, respectively. The maximal production of CMCase under optimized conditions was 83.8 U/ml, which was 3.3 times higher than that before optimization. In this study, rice hulls, agro-byproduct, were developed as a substrate for production of CMCase and time for production of CMCase was reduced to 3 days using a newly isolated marine bacterium.

• Journal of Food Hygiene and Safety
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• v.25 no.4
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• pp.367-375
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• 2010

The bacteriological and physiochemical analysis of sea water and sediments in Tongyeong harbor was conducted to evaluate sanitary conditions. The samples were collected at 8 stations established once a month from June, 2008 to May, 2009. During the study period, the range of temperature was from 6.7 to $25.2^{\circ}C$, transparency ranged from 1.2 to 2.6 m, chemical oxygen demand ranged from 1.90 to 2.92 mg/L, dissolved oxygen ranged from 6.2 to 10.5 mg/L, dissolved nitrogen ranged from 0.052 to 0.098 mg/L, phosphate ranged from 0.044 to 0.065 mg/L, respectively. Seafood, if eaten raw, carries the risk of food poisoning. Seafood poisoning is often cause by pathogenic microorganism originating from fecal contamination, such as Salmonella sp., Shigella sp. and norovirus. Fecal coliforms are an important indicator of fecal contamination. Therefore, data on fecal coliform are very important for evaluating the safety of fisheries in coastal areas. So, we investigated the sanitary indicate bacteria. The coliform group and fecal coliform MPN's of sea water in Tongyeong harbor were ranged from < 1.8~22,000/100 mL (GM 164.9 MPN/100 mL) and < 1.8~7,900 MPN/100 mL (GM 33.7 MPN/100 mL), respectively. Total coliform were detected 97.0% in 96 of samples and 68.9% of total coliforms were fecal coliforms. These results similar to another seawater detection ratio of total coloforms and fecal coliforms. The Vibrios was isolated and identified with VITEK system. Four hundred eighty strains that were obtained from sea water samples in Tongyeong harbor Detection ratio Vibrio alginolyticus, 34.2%, Vibrio parahaemolyticus, 13.8%, Vibrio vulnificus 10.0%, and V. mimicus 12.5% respectively. Vibrio cholerae O1, was not detected. During the study period, the ranges of water content, ignition loss, COD, and acid volatile sulfates in sediments in Tongyeoung harbor were 41.0~57.4%, 7.8~10.5%, 6.51~9.30 mg/g, 0.04~0.09 mg/g, respectively. Heavy metals in sediment of Tongyeoung harbor were Cd, $0.10{\pm}0.05$; Cu, $4.79{\pm}8.20$; As, $1.95{\pm}0.17$; Hg, $0.10{\pm}0.07$; $Cr^{6+}$, $0.34{\pm}0.12$; Zn, $125.33{\pm}16.40$; Ni, $16.43{\pm}1.93$ mg/kg.

• Korean Journal of Food Science and Technology
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• v.25 no.5
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• pp.449-455
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• 1993

Vibrio vulnificus is a recentry recognized halophilic organism that may cause human infections. Patients infected with Vibrio vulnificus often have a history of exposure to the sea, suggesting that the organism may be common inhabitant of marine environment. In this studies 314 samples were collected from 9 sites of Chonnam coastal area, from April to Jun, 1993. Isolation rates of V. vulnificus were 7.1% in total samples, 33.3% in sea water, 55.5% in sediment, 1.7% in fish and shellfish, 6.3% in kitchen environment. In each areas of Chunnam V. vunificus were isolated from 3.6% to 11.4%. The isolation rates of V. vulnificus was correlated positively with organic matter and COD in sea warer and sediment. In sea water, Microorganism population were $4.5{\times}10^3{\sim}3.5{\times}10^5\;CFU/ml$, Vibrio strains were $1.8{\times}10^l4.6{\times}10^3\;CFU/ml$, Coliform bacteria were $1.9{\times}10^1{\times}3.7{\times}10^2\;CFU/100ml$. In sediment, Microorganism population were $4.8{\times}10^3{\sim}5.2{\times}10^5\;CFU/ml$. Vibrio strains were $1.9{\times}10^2{\sim}8.4{\times}10^4\;CFU/ml$, Coliform bacteria were $7.2{\times}10^1{\sim}9.9{\times}10^2\;CFU/100ml$.

• Journal of Life Science
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• v.22 no.10
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• pp.1295-1306
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• 2012

A microorganism producing carboxymethylcellulase (CMCase) was isolated from seawater and identified as Bacillus atrophaeus. This species was designated as B. atrophaeus LBH-18 based on its evolutionary distance and the phylogenetic tree resulting from 16S rDNA sequencing and the neighbor-joining method. The optimal conditions for rice bran (68.1 g/l), peptone (9.1 g/l), and initial pH (7.0) of the medium for cell growth was determined by Design Expert Software based on the response surface method; conditions for production of CMCase were 55.2 g/l, 6.6 g/l, and 7.1, respectively. The optimal temperature for cell growth and the production of CMCase by B. atrophaeus LBH-18 was $30^{\circ}C$. The optimal conditions of agitation speed and aeration rate for cell growth in a 7-l bioreactor were 324 rpm and 0.9 vvm, respectively, whereas those for production of CMCase were 343 rpm and 0.6 vvm, respectively. The optimal inner pressure for cell growth and production of CMCase in a 100-l bioreactor was 0.06 MPa. Maximal production of CMCase under optimal conditions in a 100-l bioreactor was 127.5 U/ml, which was 1.32 times higher than that without an inner pressure. In this study, rice bran was developed as a carbon source for industrial scale production of CMCase by B. atrophaeus LBH-18. Reduced time for the production of CMCase from 7 to 10 days to 3 days by using a bacterial strain with submerged fermentation also resulted in increased productivity of CMCase and a decrease in its production cost.

• Clean Technology
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• v.22 no.3
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• pp.175-180
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• 2016

Tyrosinases catalyze the hydroxylation of a monophenol (monophenolase activity) and the conversion of an o-diphenol to o-quinone (diphenolase activity), which are mainly involved in the modification of tyrosine residues into 3,4-dihydroxyphenyl-alanine (DOPA) and DOPA/DOPAquinone-derived intermolecular cross-linking. Previously, we obtained a slightly acidic and cold-active tyrosinase, tyrosinase-CNK, by our recombinant protein approach. The enzyme showed optimal activity at pH 6.0 and 20 ℃ with an abnormally high monophenolase/diphenolase activity ratio and still had approximately 50% activity compared with the highest activity even in ice water. Here, we investigated reaction stability of the recombinant tyrosinase-CNK as a psychrophilic enzyme. The enzyme showed remarkable thermal stability at 0 ℃ and the activity was well conserved in repeated freeze-thaw cycles. Although water-miscible organic solvent as reaction media caused the activity decrease of tyrosinase-CNK as expected, the enzyme activity was not additionally decreased with increased concentration in organic solvents such as ethanol and acetonitrile. Also, the enzyme showed high salt tolerance in chaotropic salts. It was remarkably considered that 2⁺ metal ions might inhibit the incorporation of Cu²⁺ into the active site. We expect that these results could be used to design tyrosinase-mediated enzymatic reaction at low temperature for the production of catechols through minimizing unwanted self-oxidation and enzyme inactivation.

• Proceedings of KOSOMES biannual meeting
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• 2007.11a
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• pp.185-185
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• 2007

Starting at the beginning q the 20th century, increasing amounts of tetrach1oroethene (PCE) and trichloroethene (TCE)were manufactured due to the extensive use of these compounds in industry, in the military, and in private households, mainly as nonflammable solvents. This widespread use, along with careless handling and storage, are among the most serious contaminants of soil, sediment and groundwater. Highly chlorinated ethenes are typically not degraded through oxygenation by aerobic bacteria Since complete reductive dechlorination of PCE and TCE to ethene (ETH) has been observed in anaerobic enrichment culture, anaerobic dehalorespiring bacteria have received increased attention in the last decade. Under anaerobic conditions, these compounds con be reductively dehalogenated to less-chlorinated ethenes or innocuous ethene by microorganism through dehalorespiration. We have been studying anaerobic enrichment culture which used lactate as the electron donor for reductive dechlorination of PCE to ETH the anaerobic mixed microbial culture was enriched from the sediment sample taken from site contaminated with PCE. PCE was consistently and completely converted to ethene. In addition, the accumulation of intermediate products such as 1,2-ds-dichloroethene (cis-DCE) and vinyl chloride (VC) was observed in the anaerobic mixed microbial culture. the established dechlorinating enrichment culture was analyzed by DGGE using primers specific to DefrJ1ococcoides 16S rRNA gene sequences. In conclusion, we established the PCE dechlorinating enrichment culture and confirmed the existence of Dehalococcoides in an enrichment culture.

• Korean Chemical Engineering Research
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• v.49 no.6
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• pp.840-845
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• 2011

Degradation products of agarose are biologically active and thus used as an ingredient in pharmaceuticals or functional cosmetics. Furthermore, it has been strongly considered as a substrate for bio-ethanol fermentation. Recently, we isolated new agarase-producing microorganism, Pseudoalteromonas sp. from south sea of Korea. In this study, we aimed to separate and purify the agarase from culture broth of this strain. Separation of agarase was performed by ion- exchange chromatography on DEAE-Sepharose resin. Equilibrium pH and volume ratio of resin to the amount of protein were optimized for the efficient adsorption of protein. 410 ${\mu}g$ of protein was completely adsorbed to 3 mL of resin at pH 7.5. The total amount of eluted protein increased as NaCl concentration increased to 400 mM at isocratic elution. Agarase was separated by linear gradient elution of NaCl (0~1,000 mM). Three major protein peaks were observed and the presence or absence of agarase in these eluted proteins was measured by Lugol's staining technique. Only six eluted protein fractions showed strong agarase activity.