• Korean Journal of Microbiology
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• v.32 no.4
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• pp.285-290
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• 1994

About 500 bacterial and fungal strains from a wide variety of natural habitats were screened for a new type II restriction endonuclease. Among the 500 species, we selected one species that produced a new restriction endonuclease. This strain has an optimum temperature of $30^{circ}C$ for growth. Morphological, cultural, and physiological characteristics were examined for identification of the isolated strain J-482. This strain was found to belong to the genus Alcaligenes. The restriction endonuclease was named as AspJI and partially purified from Alcaligenes sp. J-482 by DEAE-Sephadex A-50 column chromatography and gel filtration. Most of other nucleases were removed by the purification steps. The AspJI has a substrate specificity to ${lambda}$ DNA, pBR322 and Adenovirus-2 DNA. For its maximal activity, the isolated enzyme requires $MgCl_2$, which should be at least 12.5 mM and it does not need any other cofactors. It is maximally active in the absence of NaCl and is completely inactivated at 100 mM NaCl. The pH and temperature optima for activity were pH 7.5 and $37^{circ}C$, respectively. The DNA fragments generated by digesting ${lambda}$ DNA, pBR322, and Adenovirus-2 DNA with AspJI were the same as that produced by AatII. This suggests that AspJI is an isoschizomer of AatII.

• Korean Journal of Soil Science and Fertilizer
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• v.42 no.1
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• pp.29-36
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• 2009

Fourteen bacterial strains were isolated from crop rhizosphere and identified as phosphate solubilizing bacteria (PSB) by 16S rRNA analysis. Only 3 strains exhibited a strong ability to solubilize insoluble phosphate in agar medium containing a hydroxyapatite. The rates of P solubilization by isolates were ranged from 200 and $2300\;mg\;L^{-1}$, which are inversely correlated with pH in culture medium. Furthermore, HPLC analyses reveal the production of organic acid from the culture filtrates of PSB. Among these, strain Acinetobacter sp. released only gluconic acid, Pseudomonas orientalis produced gluconic acid which was subsequently converted into 2-ketogluconic acid, and Enterobacter asburiae released acetic acid and succinic acid. On the other hand, P. orientalis and E. asburiae released $372\;mg\;L^{-1}$ and $191\;mg\;L^{-1}$ of IAA into broth culture, respectively, while Acinetobacter sp. did not produce IAA. Furthermore, in vivo study showed that plant growth promoting effect by bacteria generally seemed to be increased IAA production and phosphate solubilization.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.4
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• pp.339-347
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• 2000

An algicidal bacterium, Micrococcus sp. LG-5 against the harmful dinoflagellate, Cochlodinium polykrikoides was isolated. The optimal conditions for the highest algicidal activity of bacterial culture filtrate showed in the range of $20{\~}30^{\circ}C$, at pH 7.0 and $3.0{\%}$ of NaCl concentration. In addition, $IC_(50)(mean of 50{\%} inhibitory concentration)$ of the culture filtrate against C. polykrikoides after incubation of 5 days was $0.482{\%}$. To investigate heat and pH stability of the culture filtrate of Micrococcus sp. LG-5, the culture filtrate ($pore size, 0.1 {\mu}m$) was heated to $121^{\circ}C for 15 min$ and adjusted pH from 2.0 to 10.0. There were no significant changes in algicidal activity by heat treatment and the pH change between pH from 5.0 to 10.0. The algicidal substances produced from Micrococcus sp. LG-5 were mainly detected in the fraction of $10,000{\~}1,000$ MWCO (molecular weight cut-off). The culture filtrate of Micrococous sp. LG-5 showed antimicrobial activity against Enterococcus faecalis, Escheiichia coli, Uebsiella pneunioniae and Vibrio altinolyticus, but did not show against Pseudomonas aeminosa, P. Buorescens, Salmonella typhi, Staphylococcus aureus, V. cholerae and V parahaemolyicus. The culture filtrate of Micrococcus sp. LG-5 was examined against 16 phytoplankton species and showed the algicidal activity against Ajexandzium tuarense, Eutreptiella Drnnastin, Gymnodinium catenatum, G. mikimotoi, G. sanguineum, eyodinium impuaicum, Heterocapsa triquetra, Heterosipa akashiwo, Prorocentrum micans and Pyraminonas sp.. However no algicidal effects of Micrococcus sp. LG-5 were observed against Chlamydomonas sp., Cylindrotheoa closterium, P. mininum, P. triestimum, Pseudonieschia sp. and Sczipuiella trochoidea. On the other hand, algicidal activity on the tested marinelivefood was not detected except for Isochrysis galbana. In addition, physiological responses of cultured olive flounder (Paralichthys oliraceus) exposed to $1 and 10{\%}$ of the culture filtrate of Micrococcus sp. LG-5 were measured. There were no clear changes in AST, GGT, creatinine, urea, total cholesterol, total protein, albumine, $Mg^(+2), Ca^(+2), Na^+, K^+, and Cl^-$. These results indicate that olive flounders were not affected when they were exposed to the culture filtrate of Micrococcus sp. LG-5.

• Journal of the Korea Organic Resources Recycling Association
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• v.11 no.2
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• pp.55-65
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• 2003

The combined effect of bioaugmentation of dechlorinating bacterial cultures and addition of iron powder($Fe^0$ on reductive dechlorination of tetrachloroethylene(PCE) and other chlorinated ethylenes in a artificially contaminated soil slurry(60micromoles PCE/kg soil). Two different anaerobic bacterial cultures, a pure bacterial culture of Desulfitobacterium sp. strain Y-51 capable of dechlorinating PCE to cis-1,2-dechloroethylene(cis-DCE) and the other enrichment culture PE-1 capable of dechlorinating PCE completely to ethylene, were used for the bioaugmentation test. Both treatments introduced with the strain Y-51 and PE-1 culture (3mg dry cell weight/kg soil) showed conversion of PCE to cis-DCE within 40days. The treatments added with $Fe^0$(0.1-1.0%) alone to the soil slurry resulted in extended PCE dechlorination to ethylene and ethane and the dechlorination rate depended on the amount of $Fe^0$ added. The combined use of the bacterial cultures with $Fe^0$(0.1-1.0%)) showed the higher PCE dechlorination rate than the separated application and the pattern of PCE dechlorination and end-product formation was different from those of the separated application. When 0.1% of $Fe^0$ was added with the cultures, the treatments with the strain Y-51 and $Fe^0$ resulted in cis-DCE accumulation from PCE dechlorination, but the treatment with the enrichment culture and $Fe^0$ showed the more extended dechlorination via cis-DCE. These results suggested that the combined application of and the bactrial culture, specially the complete dechlorinating enrichment culture, is practically effective for bioremediation of PCE contaminated soil.

• Journal of Microbiology and Biotechnology
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• v.19 no.4
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• pp.339-345
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• 2009

We evaluated the activity and abundance of the crude-oil-degrading bacterium Nocardia sp. H17-1 during bioremediation of oil-contaminated soil, using real-time PCR. The total petroleum hydrocarbon(TPH) degradation rate constants(k) of the soils treated with and without H17-1 were $0.103\;d^{-1}$ and $0.028\;d^{-1}$ respectively. The degradation rate constant was 3.6 times higher in the soil with H17-1 than in the soil without H17-1. In order to detect and quantify the Nocardia sp. H17-1 in soil samples, we quantified the genes encoding 16S ribosomal RNA(16S rRNA), alkane monooxygenase(alkB4), and catechol 2,3-dioxygenase(23CAT) with real-time PCR using SYBR green. The amounts of H17-1 16S rRNA and alkB4 detected increased rapidly up to 1,000-folds for the first 10 days, and then continued to increase only slightly or leveled off. However, the abundance of the 23CAT gene detected in H17-1-treated soil, where H17-1 had neither the 23CAT gene for the degradation of aromatic hydrocarbons nor the catechol 2,3-dioxygenase activity, did not differ significantly from that of the untreated soil($\alpha$=0.05,p>0.22). These results indicated that H17-1 is a potential candidate for the bioaugmentation of alkane-contaminated soil. Overall, we evaluated the abundance and metabolic activity of the bioremediation strain H17-1 using real-time PCR, independent of cultivation.

• Food Science and Biotechnology
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• v.17 no.5
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• pp.971-975
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• 2008

This study was to examine the potential disinfection efficiencies of 10 compounds by determining their antimicrobial capacity and ichthyotoxicity. Antimicrobial effects against Vibrio sp., Edwadsiella tarda, Streptococcus sp., and Staphylococcus sp. were tested using 10 different disinfectants; hydrogen peroxide, sodium hypochlorite, chlorine dioxide, povidon iodine, formaldehyde, glutaraldehyde, quaternary ammonium compounds (QACs), didecyl dimethyl ammonium chloride (DDAC), ortho-dichlorobenzen, and copper sulfate. Chlorine dioxide ($ClO_2$) containing 5% $ClO_2$ and copper sulfate had no effects on bactericidal activity, while the other disinfectants resulted in 99.99% bactericidal activity against 4 strains of fish pathogenic bacteria. The ichthyotoxicity of the 10 disinfectants was investigated using 3 kinds of fish species; flounder (Paralichthys olivaceus), rockfish (Sebastes pachycephalus), and black sea bream (Acanthopagrus schlegelii). Median lethal concentration ($LC_{50}$) values of the 10 disinfectants were estimated to determine toxicity ranges of the doses within 24 hr. Among test disinfectant solutions, hydrogen peroxide showed the highest $LC_50$ in flounder (201.3), rockfish (269.7), and black sea bream (139.3 ppm). DDAC revealed the lowest $LC_{50}$ in flounder (2.1), rockfish (1.0), and black sea bream (1.5 ppm). These results suggest that DDAC, quaternary ammonium compounds, glutaraldehyde, and sodium hypochlorite are effective disinfectants for fish and bacterial species examined in this study.

• Journal of the Korean Society of Food Culture
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• v.27 no.6
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• pp.743-750
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• 2012

A tissue cultured wild ginseng (TCWG) suspension was inoculated with lactic acid bacteria and fermented to improve the functionality of TCWG. The utilization of TCWG was increased directly using the freeze-dried powder. The optimal ratio of TCWG powder and water for fermentation was 1:19 (5%), which was selected by measuring the fluidity and viable cell count according to concentration. The effects on ADH activation and immune cell activation by each ferments with 10 kinds of Lactobacillus sp. strains were examined. The ferments with the Lactobacillus casei KFRI 692 strain showed 5.4 times higher ADH activity and 1.3 times higher ALDH activity than the non-fermented TCWG powder (control). The level of NO production and cytotoxicity was also measured by Raw 264.7 cells. The ferment with the Lac. casei KFRI 692 strain showed the highest level of NO production and lower cytotoxicity than the others. Therefore, the Lac. casei KFRI 692 strain was selected as a strain for fermentation of a TCWG suspension to maximize its functionality. To identify the optimal fermentation time of the selected Lac. casei KFRI 692 strain on the 5% TCWG suspension, the viable cell count of lactic acid bacterial and the changes in pH were observed for 72 hours. 24-hrs was found to be the optimal fermentation time. In this way, fermented TCWG with lactic acid bacteria showed higher ADH activation efficacy and immune cell activation than non-fermented TCWG.

• International Journal of Industrial Entomology and Biomaterials
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• v.25 no.2
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• pp.195-203
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• 2012

In reared populations of Allomyrina dichotoma, commercial insects, the skin of last instar larvae was changed softer with opaque white, and infested grubs eventually died. To clarify the cause of the symptom, we collected the larvae of A. dichotoma from five farms and examined their intestinal bacterial florae using pyrosequencing technique. From those results, a member of Paenibacillus was found only in the larvae showing the symptom of disease. Through PCR analysis using a Paenibacillus specific primer set, we obtained the partial 16S rRNA gene sequence and confirmed the microbe as Paenibacillus sp. For clear identification, a whole guts was extracted from each larva showing the sign of the disease and incubated at $70^{\circ}C$ for 15 min to isolate spore forming bacteria. After then, each content of guts was cultured on $MYPGP_{NAL}$ agar medium($12.5{\mu}g/ml$ of nalidixic acid) at $30^{\circ}C$. The 16S rRNA gene sequence analysis for the isolated bacteria showed that they were closely related to P. rigui(97.9% similarity), to P. chinjuensis(96.1% similarity), and to P. soli(95.3% similarity). Additional tests including API test and cellular fatty acid composition analysis were performed, but the strain couldn't be identified at species level, suggesting it may represent novel species of the genus Paenibacillus.

• Journal of Microbiology and Biotechnology
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• v.24 no.5
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• pp.585-591
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• 2014

To evaluate the potential utility of pretreatment of raw biomass with a complex microbial system, we investigated the degradation of rice straw by BMC-9, a lignocellulose decomposition strain obtained from a biogas slurry compost environment. The degradation characteristics and corresponding changes in the bacterial community were assessed. The results showed that rapid degradation occurred from day 0 to day 9, with a peak total biomass bacterium concentration of $3.3{\times}10^8$ copies/ml on day 1. The pH of the fermentation broth declined initially and then increased, and the mass of rice straw decreased steadily. The highest concentrations of volatile fatty acid contents (0.291 mg/l lactic acid, 0.31 mg/l formic acid, 1.93 mg/l acetic acid, and 0.73 mg/l propionic acid) as well as the highest xylanse activity (1.79 U/ml) and carboxymethyl cellulase activity (0.37 U/ml) occurred on day 9. The greatest diversity among the microbial community also occurred on day 9, with the presence of bacteria belonging to Clostridium sp., Bacillus sp., and Geobacillus sp. Together, our results indicate that BMC-9 has a strong ability to rapidly degrade the lignocelluloses of rice straw under relatively inexpensive conditions, and the optimum fermentation time is 9 days.

• Journal of Microbiology and Biotechnology
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• v.18 no.6
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• pp.1011-1015
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• 2008

A Gram-negative, strictly aerobic, motile bacterial strain, designated Gsoil $124^T$, was isolated from a soil sample taken from a ginseng field in Pocheon Province (South Korea). The isolate contained Q-10 as the predominant lipoquinone, plus $C_{18:1}\;{\omega}7c$ and summed feature 4 ($C_{16:1}\;{\omega}6c$ and/or iso-$C_{15:0}$ 2-OH) as the major fatty acids. The G+C content of the genomic DNA was 68.1 mol%, and the major polar lipids consisted of sphingoglycolipid, phosphatidylglycerol, phosphatidylcholine, and phosphatidylethanolamine. A comparative 16S rRNA gene sequence analysis showed that strain Gsoil $124^T$ was most closely related to Sphingopyxis chilensis (98.7%), Sphingopyxis alaskensis (98.2%), Sphingopyxis witflariensis (98.2%), Sphingopyxis taejonensis (98.0%), and Sphingopyxis macrogoltabida (97.6%). However, the DNA-DNA relatedness between strain Gsoil $124^T$ and its phylogenetically closest neighbors was less than 22%. Thus, on the basis of its phenotypic properties and phylogenetic distinctiveness, strain Gsoil $124^T$ should be classified as representing a novel species in the genus Sphingopyxis, for which the name Sphingopyxis panaciterrae sp. nov. is proposed. The type strain is Gsoil $124^T$ (=KCTC $12580^T$=LMG $24003^T$).