• Fisheries and Aquatic Sciences
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• v.6 no.1
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• pp.27-33
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• 2003

A lactic acid bacterium showing antimicrobial activity against fish pathogen was isolated from gastrointestinal tract of flounder for the purpose of use as an aquaculture probiotics. From the analysis of morphological and physiological characteristics, the isolated strain was named as Lactococcus sp. HM58. Antimicrobial substance (AMS) from Lactococcus sp. HM58 showed strong growth inhibitory activity against Streptococcus sp., which is a fish pathogenic bacterium. AMS was presumed a proteinaceous compound with stability in heat and wide pH range from 2 to 10. It was started to produce in exponential growth phase and was not produced any more in stationary phase. It showed comparatively broad antimicrobial spectrum against most of gram positive bacteria used for this study. About $84\%$ of Lactococcus sp. HM58 was able to survive in the artificial gastric juice though it was low to the extent in the artificial bile juice. In the sensitivity test for various antibiotics, this strain was highly sensitive for doxycycline, erythromycin, amoxicillin clavu1anic acid and ampicillin.

• Proceedings of the Microbiological Society of Korea Conference
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• 2008.05a
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• pp.135-135
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• 2008

Various biosensors were evaluated for identifying and detecting foodborne pathogens in a rapid and effective manner. First, five strains of Escherichia coli and six strains of Salmonella were identified using Fourier transform infrared spectroscopy and a statistical program. For doing this, lipopolysaccharides (LPSs) and outer membrane proteins (OMPs) were extracted from a cell wall of each bacterial strain. As a result, each strain was identifed at the level of 97% for E. coli and 100% for Salmonella. Second, E. coli O157:H7, S. Enteritidis, and Listeria monocytogenes were identified by multiplex PCR products from four specific genes of each bacteria using a capillary electrophoresis (CE). Also, ground beef for E. coli O157:H7, lettuce for S. Enteritidis, and hot dog for L. monocytogenes were used to determine the possibility of detecting pathogens in foods. Foods inoculated with respective pathogen were cultivated for six hours and multiplex PCR products were obtained and assessed. The minimum detection levels of tested bacteria were <10 cells/g, <10 cells/g, and $10^4$ cells/g for E. coli O157:H7, S. Enteritidis, and L. monocytogenes, respectively. Third, it was possible to detect S. Typhimurium in a pure culture and lettuce by a bioluminescence-based detection assay using both recombinant bacteriophage P22::luxI and a bioluminescent bioreporter. In addition, bacteriophage T4 was quantitatively monitored using E. coli including luxCDABE genes.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.2
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• pp.105-111
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• 2002

The biological removal of 2,4,6-trinitrotoluene (TNT) was studied in a bench-scale bioreactor using a bacterial culture of strain OK-5 originally Isolated from soil samples contaminated with TNT. The TNT was completely removed within 4 days of incubation in a 2.5 L bench-scale bioreactor containing a newly developed medium. The TNT was catabolized in the presence of different supplemented carbons. Only minimal growth was observed in the killed controls and cultures that only received TNT during the incubation period. This catabolism was affected by the concentration ratio of the substrate to the biomass. The addition of various nitrogen sources produced a delayed effect for the TNT degradation. Tween 80 enhanced the degradation of TNT under these conditions. Two metabolic intermediates were detected and identified as 2-amino-4, 6-dinitrotoluene and 4-amino-2, 6-dinitrotoluene based on HPLC and GC-MS analyses, respectively. Strain OK-5 was characterized using the BIOLOG system and fatty acid profile produced by a microbial identification system equipped with a Hewlett Packard HP 5890 II gas chromatograph. As such, the bacterium was identified as a Stenotrophomonas species and designated as Stenotrophomonas sp. OK-5.

• Korean Journal of Microbiology
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• v.55 no.2
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• pp.174-176
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• 2019

A novel bacterium, designated strain RR4-38 (= KCTC 52651 = DSM 108068), belonging to the family Flavobacteriaceae was isolated from a biofilter in the seawater recirculating aquaculture system in South Korea. A single complete genome contig which is 3,182,272 bp with 41.9% G+C content was generated using PacBio RS II platform. The genome includes 2,829 protein-coding genes, 6 rRNA genes, 38 tRNA genes, 4 non-coding RNA genes, and 9 pseudogenes. The results will provide insights for understanding microbial activity in the seawater recirculating aquaculture system.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.40-44
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• 2003

Soil-borne infectious disease including Pythium aphanidermatum and Rhizoctonia solani causes severe damage to plants, such as cucumber. This soil-borne infectious disease was not controlled effectively by chemical pesticide. Since these diseases spread through the soil, chemical agents are usually ineffective. Instead, biological control, including antagonistic microbe can be used as a preferred control method. An efficient method was developed to select an antagonistic strain to be used as a biological control agent strain. In this new method, surface tension reduction potential of an isolate was included in the ‘decision factor’ in addition to the other factors, such as growth rate, and pathogen inhibition rate. Considering these 3 decision factors by a statistical method, an isolate from soil was selected and was identified as Bacillus sp. GB16. In the pot test, this strain showed the best performance among the isolated strains. The lowest disease incidence rate and fastest seed growth was observed when Bacillus sp. GB16 was used. Therefore this strain was considered as plant growth promoting rhizobacteria (PGPR). The action of surface tension reducing component was deduced as the enhancement of wetting, spreading, and residing of antagonistic strain in the rhizosphere. This result showed that new selection method was significantly effective in selecting the best antagonistic strain for biological control of soil-borne infectious plant pathogen. The antifungal substances against P. aphanidermatum and R. solani were partially purified from the culture filtrates of Bacillus sp. GB16. In this study, lipopeptide possessing antifungal activity was isolated from Bacillus sp. GB16 cultures by various purification procedures and was identified as a surfactin-like lipopeptide based on the Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), high performance liquid chromatography mass spectroscopy (HPLC-MS), and quadrupole time-of-flight (Q-TOF) ESI-MS/MS data. The lipopeptide, named GB16-BS, completely inhibited the growth of Pythium aphanidermatum, Rhizoctonia solani, Penicillium sp., and Botrytis cineria at concentrations of 10 and 50 mg/L, respectively. A novel method to prevent the foaming and to provide oxygen was developed. During the production of surface active agent, such as lipopeptide (surfactin), large amount of foam was produced by aeration. This resulted in the carryover of cells to the outside of the fermentor, which leads to the significant loss of cells. Instead of using cell-toxic antifoaming agents, low amount of hydrogen peroxide was added. Catalase produced by cells converted hydrogen peroxide into oxygen and water. Also addition of corn oil as an oxygen vector as well as antifoaming agent was attempted. In addition, Ca-stearate, a metal soap, was added to enhance the antifoam activity of com oil. These methods could prevent the foaming significantly and maintained high dissolved oxygen in spite of lower aeration and agitation. Using these methods, high cell density, could be achieved with increased lipopeptide productivity. In conclusion to produce an effective biological control agent for soil-borne infectious disease, following strategies were attempted i) effective screening of antagonist by including surface tension as an important decision factor ii) identification of antifungal compound produced from the isolated strain iii) novel oxygenation by $H_2O_2-catalase$ with vegetable oil for antifungal lipopeptide production.

• Applied Biological Chemistry
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• v.13 no.2
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• pp.111-129
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• 1970

As a series of studies on the nucleic acids and their related substances 210 samples were collected from 76 places such as farm soil, compost of heap, nuruk and meju to obtain microbial strains which produce 5'-phosphodiesterase. From these samples total of 758 strains were isolated by the use of dilution pour plate method. For all isolated strains primary screening of the productivity of RNA depolymerase was performed and useful strains with regard to 5'-phosphodiesterase productivities were identified. For these useful strains optimum condition, the effect of various compounds on the activity of 5'-phosphodiesterase, and the optimum condition for enzyme reaction were discussed. The quantitative of 5'-mononucleotides produced by the action of 5'-phosphodiesterase was performed using anion-exchange column chromatography and their identified was done by paper chromatography, thinlayer chromatography, ultra violet spectrophotometry, and characteristic color reaction using carbazole and schiff's reagent. (1) Penicillium citreo-viride PO 2-11 and Streptomyces aureus SOA 4-21 from soil were identified as a potent 5'-phosphodiesterase producing strains. (2) Optimum culture conditions for Penicillium citreo-viride PO 2-11 strain isolated were found to be pH 5.0 and $30^{\circ}C$, and the optimum conditions for enzyme action of 5'-phosphodiesterase were pH 4.2 and $60^{\circ}C$. Best carbon source for the production of 5'-phosphodiesterase was found to be sucrose and ammonium nitrate for nitrogen source. Addition of 0.01% corn steep liquor or yeast extract exhibited 20% increase in the amount of 5'-phosphodiesterase production compared to the control. 5'-phosphodiesterase produced by this strain was activated by $Mg^{++},\;Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by EDTA, citrate, $Cu^{++},\;CO^{++}$. 5'-phosphodiesterase produced 5'-mononucleotide from RNA at a rate of 65.81%, and among the 5'-mononucleotides accumulated 5'-GMP only was found to have flavorous and the strain was also found lack of 5'-AMP deaminase. Productivity of flavorous 5'-GMP was found to be 186.7mg per gram of RNA. (3) Optimum culture canditions for the isolated Streptomyces aureus SOA 4-21 strain were pH 7.0 and $28^{\circ}C$, and the optimum conditions for the action of 5'-phosphodiesterase were pH 7.3 and $50^{\circ}C$. The best carbon source for 5'-phosphodiesterase production was found to be glucose and that of nitrogen was asparagine. Addition of 0.01% yeast extract exhibited increased productivity of 5'-phosphodiesterase by 40% compared to the non-added control. 5'-phosphodiesterase produced by this strain was activated by $Ca^{++},\;Zn^{++},\;Mn^{++}$ and was inhibited by citrate, EDTA, $Cu^{++}$. It was also found that the strain produce 5'-AMP deaminase in addition to 5'-phosphodiesterase. For this reason although decomposition rate was 63.58% the accumulation of 5'-AMP, 5'-CMP, 5'-GMP and 5'-UMP occurred by the breakdown of RNA. In the course of these reaction 5'-AMP deaminase converted 60% of 5'-AMP thus produced into 5'-IMP and flavorous 5'-mono nucleotide production was significantly increased by this strain over the above mentioned one. Production rates were found to be 171.8mg per grain of RNA for 5'-IMP and 148.2mg per gram of RNA for 5'-GMP, respectively.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.95.2-96
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• 2003

In order to develop a new microbial fungicide for the control of vegetable diseases using endophytic bacteria, a total of 260 bacterial strains were isolated from fresh tissues of 5 plant species. After they were cultured in broth media, their antifungal activities were screened by in vivo bioassays against Botrytis cinerea(tomato gray mold), Pythium ultimum(cucumber damping-off), Phytopkhora infestans(tomato late blight), Colletotrichum orbiculare(cucumber anthracnose), and Blumeria graminis f. sp. hordei(barley powdery mildew). As the results of screening, 38 bacterial strains showed potent antifungal activities against at least one of 5 plant pathogens. A bacterial strain EB072 displayed potent disease control activities against 3 plant diseases. Among the bacterial strains with a potent antifungal activity against cucunlber anthracnose, three bacterial strains, EB054, EB151 and EB215, also displayed a potent in vitro antifungal activity against C. acutatum, a fungal agent causing pepper anthracnose. A bacterial strain EB215 obtained from roots of cucumber was identified as Burkholderia cepacia based on its physiological and biochemical characteristics and 165 rRNA gene sequence. An antifungal substance was isolated from the liquid cultures of B. cepacia EB215 strain by ethyl acetate partitioning, repeated silica gel column chromatography, and invitro bioassay, Its structural determination is in progress by various instrumental analyses.

• Applied Biological Chemistry
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• v.46 no.1
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• pp.1-6
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• 2003

Abstract: In order to improve the system for biological nitrogen oxidizing process in sewage and wastewater, a bacterium having high abilities to oxidize of nitrogen was isolated from wastewater and polluted soils. The strain was identified to Bacillus sp. CH-N, based on the physiological and biochemical properties. Characteristics and oxidizing ability of both ammonia and nitrite were examined for the strain, Bacillus sp. CH-N. The strain showed the oxidizing rate about 80% to 90% on the sewage and wastewater after 48 h culture. The nitrogen oxidizing rate was increased in proportion to the initial concentration of glucose. The microorganism, Bacillus sp. CH-N cell immobilized on ceramic carrier were evaluated for the oxidation of ammonia in culture media.

• Journal of Microbiology and Biotechnology
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• v.15 no.4
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• pp.716-721
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• 2005

The role of an effective microbial species is critical to the successful application of biological processes to remove sulfur compounds. A bacterial strain was isolated from the soil of a malodorous site and identified as Burkholderia spp. This isolate was able to oxidize thiosulfate to sulfate, with simultaneous pH decrease and accumulation of elemental sulfur. The specific growth rate and the sulfate oxidation rate using the thiosulfate basal medium were $0.003 h^{-1}\;and\;3.7 h^{-1}$, respectively. The isolated strain was mixotrophic, and supplementation of $0.2\%$ (w/v) of yeast extract to the thiosulfate-basal medium increased the specific growth rate by 50-fold. However, the rate of sulfate oxidation was more than ten times higher without yeast extract. The isolate grew best at pH 7.0 and $30^{\circ}C$, and the sulfate oxidation rate was the highest at 0.12 M sodium thiosulfate. In an upflow biofilter, the isolated strain was able to degrade $H_2S\;with\;88\%$ efficiency at 8 ppm and 121/h of incoming gas concentration and flow rate, respectively. The cell density at the bottom of the column reached $3.2{\times}10^8$ CFU/(g bead) at a gas flow rate of 121/h.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.457-460
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• 2002

In an effort to isolate novel strains expressing a thermostable esterase that hydrolyzed the rac-ketoprofen ethyl ester to ketoprofen in the stereospecific manner, we screened various soils and composts from broad ecological niches in which the activity was expected to be found. Three hundreds of microbial strains were tested to determine their ester-hydrolyzing activity by using an agar plate containing insoluble tributyrin as an indicative substrate, and then further screened by activity on the (R,S)-ketoprofen ethyl ester. Twenty-six strains were screened primarily at high growth and incubation temperature and further compared the ability to ethyl ester-hydrolyzing activity in terms of conversion yield and chiral specificity. Consequently, a strain JYl44 was isolated as a novel strain that produced a (R)-stereospecific esterase with high stability and systematically identified as a Bacillus stearothermophilus JY144. The enzyme indeed stables at a broad range of temperature, upto 65 $^{\circ}C$, and pH ranging from 6.0 to 10.0. The optimal temperature and pH for enzymatic conversion were 50 $^{\circ}C$ and 9.0, respectively. Based on the observations that resulted a poor cell growth, and enzyme expression in wild type strain, we further attempted the gene cloning into a general host Escherichia coli and determined its primary structure, concomitantly resulting a high level expression of the enzyme. The cloned gene had an open reading frame (250 amino acids) with a calculated molecular mass of 27.4 kDa, and its primary structure showed a relative high homology (45-52 %) to the esterases from Streptomyces and Bacillus strains. The recombinant whole cell enzyme could efficiently convert the rac-ketoprofen ethyl ester to (R)-ketoprofen, with optical purity of 99 % and yield of 49 %.