• Journal of Microbiology and Biotechnology
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• v.10 no.2
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• pp.258-263
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• 2000

2-Hydroxy-6-oxo-6phenylhexa-2,4-dienoate (HOPDA) hydrolase catalyzes the hydrolytic cleavage of HOPDA to bemzpate and 2-hydroxypenta-2, 4-dienoate (HPD) during microbial catabolism of biphenyl and polychlorinated biphenyls. A HOPDA hydrolase gene (pcbD) was isolated from the genomic library of Pseudomonas sp. P20 and designated as pCNUO1201; a 7.5-kb XbaI DNA fragment from Pseudomonas sp. P20 was inserted into the pBluescript SK(+) XbaI site. E. coli HB101 harboring pCNU1201 exhibited HOPDA hydrolase activity. The open reading frame (ORF) corresponding to the pcbD gene consisted of 855 base pairs with an ATG initiation codon and a TGA termination codon. The ORF was preceded by a rebosome-binding sequence of 5'-TGGAGC-3' and its G+C content was 55 mol%. The pcbD gene of Pseudomonas sp. P20 was located immedeately downstream of the pcbC gene encoding 2,3- dihydroxybiphenyl 1,2-dioxygenase, and approximately 4-kb upstream of the pcbE gene encoding HPD hydratase. The pcbK gene was able to encode a polypeptide with a molecular weight of 31,732 containing 284 amino acid residues. The deduced amino acid sequence of the HOPDA hydrolase of Pseudomonas sp. P20 exhibited high identity (62%) with those of the HOPDA hydrolases of P. putida KF715, P. pseudoalcaligenes KF707, and Burkholderia cepacia LB400, and also significant homology with those of other hydrolytic enzymes including esterase, transferase, and peptidase.

• The Journal of the Korean Society for Microbiology
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• v.16 no.1
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• pp.29-34
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• 1981

Species and antimicrobial susceptibility of nonfermenting gram-negative bacilli(NFB) isolated from clinical materials at Yonsei Medical Center during the period of September 1980 to August 1981 were analyzed and the following results were obtained. 1. NFB were isolated from 17.1% of sputum, 14.8% of pus, 5.0% of urine, 3.3% of throat and 1.4% of blood specimens. 2. Among the isolates 57.6% were Pseudomonas aeruginosa and 32.7% were Acinetobacter calcoaceticus. P. maltophilia and P. cepacia were 3.2% and 2.8% respectively. Other bacteria were rarely isolated. 3. The monthly isolation rates were high during the June to November period(8.8-12.0%), and low during the December to May period(4.2-8.4%). 4. Many of the isolates showed resistance to various antimicrobials. Although there were variations depending on the bacterial species, amikacin, colistin, gentamicin, tobramycin and co-trimoxazole showed more antibacterial activities than other antimicrobials.

• Korean Journal of Microbiology
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• v.51 no.3
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• pp.271-279
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• 2015

Lactoferrin (LF) is a multifunctional, iron-binding glycoprotein found in physiological secretions of mammals. LF shows antibacterial, antiviral and antifungal activities. In the present study, a gene encoding the N-terminal lobe of human lactoferrin (hLF) was isolated, cloned and expressed in methylotrophic yeast, Pichia pastoris. The recombinant hLF-N (rhLF-N) protein was secreted into the culture medium at the level of $458{\mu}g/ml$ in 3 L fermentor. The size of purified hLF-N was estimated as 35 kDa when analyzed by SDS-PAGE and western blotting. The rhLF-N was further confirmed by immunodiffusion using the anti-hLF polyclonal antibody. The expression profile analysis by qRT-PCR showed that the relative mRNA expression of rhLF-N was maximal after 2-3 days of methanol induction and reduced gradually at 4 days. The purified rhLF-N showed broad antibacterial activities against the pathogens such as Staphylococcus aureus, E. coli, Pseudomonas aeruginosa, Burkholderia cepacia, and Salmonella typhimurium. However, rhLF-N showed relatively lower activity when compared to peptides derived from LF. In spite of this weak activity, the rhLF-N expressed in P. pastoris might be more advantageous for the industrial application, because rhLF-N is secreted into the culture medium and the production can also be increased by optimization of culture conditions.

• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.376-383
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• 2005

Alcaligenes sp. JMP228 carrying 2,4­dichlorophenoxyacetic acid (2,4-D) degradative plasmid pJP4 was inoculated into natural soil, and transfer of the plasmid pJP4 to indigenous soil bacteria was investigated with and without 2,4-D amendment. Plasmid pJP4 transfer was enhanced in the soils treated with 2,4-D, compared to the soils not amended with 2,4-D. Several different transconjugants were isolated from the soils treated with 2,4-D, while no indigenous transconjugants were obtained from the unamended soils. Inoculation of the soils with both the donor Alcaligenes sp. JMP228/pJP4 and a recipient Burkholderia cepacia DBO 1 produced less diverse transconjugants than the soils inoculated with the donor alone. Repetitive extragenic palindromic-polymerase chain reaction (REP-PCR) analysis of the transconjugants exhibited seven distinct genomic DNA fingerprints. Analysis of 16S rDNA sequences indicated that the transconjugants were related to members of the genera Burkholderia and Pandoraea. Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA genes revealed that inoculation of the donor caused clear changes in the bacterial community structure of the 2,4-D­amended soils. The new 16S rRNA gene bands in the DGGE profile corresponded with the 16S rRNA genes of 2,4-D­degrading transconjugants isolated from the soil. The results indicate that introduction of the 2,4-D degradative plasmid as Alcaligenes sp. JMP228/pJP4 has a substantial impact on the bacterial community structure in the 2,4-D-amended soil.

• Journal of the Korean Society of Industry Convergence
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• v.2 no.1
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• pp.85-90
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• 1999

The possibility of employment of ceramics, known to be emitting far infrared waves, to keep the growth and the viability of the antagonistic microorganisms was examined. Among four kinds of commercial ceramics, the ceramic powders composed of soft ferrite as a main component has exhibited to grow Pseudomonas cepacia and Saccharomyces cerevisiae better. The ceramic powder in the growth medium has increased the number of cells of P. cepada about ten times and that of S. cerevisiae two to five times more than that in the control at $27^{\circ}C$. The viability of the microorganism at low temperature which was measured from the regrowth behavior at $27^{\circ}C$ after five days store at $4^{\circ}C$ has shown that the lag time of the two microorganisms reduced about three hours without any defect in the rate of logarithmic growth. These results demonstrated that the ceramic powders was available to the growth and viability at low temperature of antagonistic microorganisms.

• Microbiology and Biotechnology Letters
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• v.31 no.4
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• pp.311-321
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• 2003

Bacteria produce lipases, which can catalyze both the hydrolysis and the synthesis of long chain triglycerides. These reactions usually proceed with high regioselectivity and enantioselectivity, and, therefore, lipases have become very important biocatalysts used in organic chemistry. 3D lipase structures were solved from several bacterial lipases. They have an $\alpha/\beta$ hydrolase fold and a catalytic triad consisting of a nucleophilic serine, and an aspartate or glutamate residue that is hydrogen bonded to a histindine. Active sites are covered with $\alpha$-helical lid structure, of which movement is involved in the enzyme's activation at oil/water interface. Four substrate binding pockets were identified for triglycerides: an oxyanion hole and three pockets accommodating the fatty acids bound at positions sn-1, sn-2, and sn-3. These pockets determine the enantiopreference of a lipase. The understanding of structure-function relationships as well as the development of molecular evolution techniques will enable researchers to tailor new lipases for biotechnological applications.

• Korean Journal of Soil Science and Fertilizer
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• v.35 no.2
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• pp.118-126
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• 2002

To evaluate the correlations of microbial populations with soil healthiness and crop production and establish the criteria for microbial population of soil types. We analyzed the microbial community structure of 13 soils which were different in physical and chemical properties and cultivation methods. According to the analysis of microbial population suing the dilution plate method, the large differences of the microbial population structures among soil types were shown: aerobic bacteria $2-27{\times}10^6$, fluorescent Pseudomonas $1-1,364{\times}10^5$, Gram negative bacteria $1-126{\times}10^4$, and mesophilic Bacillus $1-110{\times}10^5$. The density of Gram negative bacteria was highest on red pepper cultivating soils (sample no. 4 and 6) of Umsung and Gesan, Chungbuk, and the density of the fluorescent Pseudomonas was highest on greenhouse soil (sample no. 7) of Jinju, Kyungnam. The crop productivity of three soils was high as compared with those of other soils. It was supposed that the density of fluorescent Pseudomonas and mesophilic Bacillus were correlated with the incresed crop production. By MIDI analysis, 579 strains isolated from 13 soils composed of a variety of microbes including 102 isolates of Agrobacterium, 112 isolates of Bacillus, 32 isolates of Pseudomonas, 44 isolates of Kocuria, and 34 isolates of Pseudomonas. Among the 624 isolates of Gram negative bacteria, Pseudomonas including P. putida and p. fluorescens occupied the highest density (51%), and Stenotrophomonas maltophilia and Burkholderia cepacia also appeared at high density. From RAPD analysis, the fluorescent Pseudomonas strains isolated from 13 soil types showed a high level of strain diversities and were grouped into 2 - 14 patterns according to soil types. Many of unknown bacteria were recovered from the paddy soil, and needed to be further characterized on the molecular basis.

• Pediatric Infection and Vaccine
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• v.10 no.2
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• pp.167-177
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• 2003

Purpose : To know the trends of antimicrobial susceptibility is critical for antimicrobial treatment. We studied the organisms isolated from blood, urine, stool, and cerebrospinal fluid from 1997 to 2001 to reveal the trends of their antimicrobial susceptibility. Methods : We conducted a retrospective study with isolates obtained from 0~18 year old outpatients and inpatients from 1997 to 2001 at Department of Pediatrics, Hanil general hospital. We gathered the data through the laboratory test files and the origin of microorganisms cultured from blood, urine, stool and cerebrospinal fluid and their antimicrobial susceptibility. Results : Microorganisms were isolated from 226(3.3%) out of 6,974 blood cultures, 365 (8.0%) out of 4,549 urine cultures, 50(1.9%) out of 2,593 stool cultures and 9(1.4%) in 655 cerebrospinal fluid cultures. The most frequently isolated organisms from blood cultures was Staphylococcus epidermidis(33.5%) which was followed by Staphylococcus aureus(19.7%), Escherichia coli(13.8%), and Burkholderia cepacia(9.0%). Among the urine cultures, E. coli was the most common(74.7%) which was followed by Group D Enterococcus(11.3%), Klebsiella pneumoniae(7.1%) and Proteus mirabilis(2.5%). The positive stool cultures all yield Salmonella species. Group D Salmonella was obtained most frequently. Among the positive cerebrospinal fluid cultures, Group B Streptococcus was isolated most frequently. Among the 40 cases of S. aureus in blood cultures, 27 cases were methicillin-resistant. The rates of susceptibility for amikacin, ceftizoxime and ceftriaxone of E. coli isolated from blood cultures were 80%, 100% and 60% in 1997 and 60%, 80% and 60% in 2001. The rates of susceptibility for amikacin, ceftizoxime and ceftriaxone of K. pnumoniae isolated from urine cultures. were 80%, 100% and 80% in 1997 and 50%, 83% and 50% in 2001 Enterococcus was isolated from 6.7% to 15.8% and vancomycin-resistant Enterococcus was observed in 17% of Group D Enterococcus isolated from urine cultures. The rates of susceptibility for amikacin, ceftizoxime and ceftriaxone of Group D Salmonella were 96%, 96% and 92% during the study period. Conclusion : Among the blood cultures S. epidermidis, S. aureus, E. coli and B. cepacia were isolated in order of frequency and among the urine cultures E. coli, Group D Enterococcus, K. pneumoniae and P. mirabilis were isolated in order of frequency. During the study period there was no big difference in major organisms isolated from blood and urine. The methicillin-resistant S. aureus was observed in 67% of S. aureus isolated from blood cultures but vancomycin-reistant S. aureus or vancomycin intermediate resistant S. aureus was not observed. The rates of susceptibility to amikacin and the third generation cephalosporin of E. coli isolated from blood cultures and K. pneumoniae from urine cultures have decreased. The isolation rates of Group D Enterococcus and vancomycin resistant Enterococcus have increased.

• Fisheries and Aquatic Sciences
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• v.21 no.2
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• pp.6.1-6.10
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• 2018

The intention of this study was to identify the bacterial pathogens infecting Oreochromis niloticus (Nile tilapia) and Clarias gariepinus (African catfish), and to establish the antibiotic susceptibility of fish bacteria in Uganda. A total of 288 fish samples from 40 fish farms (ponds, cages, and tanks) and 8 wild water sites were aseptically collected and bacteria isolated from the head kidney, liver, brain and spleen. The isolates were identified by their morphological characteristics, conventional biochemical tests and Analytical Profile Index test kits. Antibiotic susceptibility of selected bacteria was determined by the Kirby-Bauer disc diffusion method. The following well-known fish pathogens were identified at a farm prevalence of; Aeromonas hydrophila (43.8%), Aeromonas sobria (20.8%), Edwardsiella tarda (8.3%), Flavobacterium spp. (4.2%) and Streptococcus spp. (6.3%). Other bacteria with varying significance as fish pathogens were also identified including Plesiomonas shigelloides (25.0%), Chryseobacterium indoligenes (12.5%), Pseudomonas fluorescens (10.4%), Pseudomonas aeruginosa (4.2%), Pseudomonas stutzeri (2.1%), Vibrio cholerae (10.4%), Proteus spp. (6.3%), Citrobacter spp. (4.2%), Klebsiella spp. (4.2%) Serratia marcescens (4.2%), Burkholderia cepacia (2.1%), Comamonas testosteroni (8.3%) and Ralstonia picketti (2.1%). Aeromonas spp., Edwardsiella tarda and Streptococcus spp. were commonly isolated from diseased fish. Aeromonas spp. (n = 82) and Plesiomonas shigelloides (n = 73) were evaluated for antibiotic susceptibility. All isolates tested were susceptible to at-least ten (10) of the fourteen antibiotics evaluated. High levels of resistance were however expressed by all isolates to penicillin, oxacillin and ampicillin. This observed resistance is most probably intrinsic to those bacteria, suggesting minimal levels of acquired antibiotic resistance in fish bacteria from the study area. To our knowledge, this is the first study to establish the occurrence of several bacteria species infecting fish; and to determine antibiotic susceptibility of fish bacteria in Uganda. The current study provides baseline information for future reference and fish disease management in the country.

• Food Science of Animal Resources
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• v.26 no.3
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• pp.402-409
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• 2006

Although microorganisms are, in fact, the most diverse and abundant type of organism on Earth, the ecological functions of microbial populations remains poorly understood. A variety of bacteria including marine Vibrios encounter numerous ecological challenges, such as UV light, predation, competition, and seasonal variations in seawater including pH, salinity, nutrient levels, temperature and so forth. In order to survive and proliferate under variable conditions, they have to develop elaborate means of communication to meet the challenges to which they are exposed. In bacteria, a range of biological functions have recently been found to be regulated by a population density-dependent cell-cell signaling mechanism known as quorum-sensing (QS). In other words, bacterial cells sense population density by monitoring the presence of self-produced extracellular autoinducers (AI). N-acylhomoserine lactone (AHL)-dependent quorum-sensing was first discovered in two luminescent marine bacteria, Vibrio fischeri and Vibrio harveyi. The LuxI/R system of V. fischeriis the paradigm of Gram-negative quorum-sensing systems. At high population density, the accumulated signalstrigger the expression of target genes and thereby initiate a new set of biological activities. Several QS systems have been identified so far. Among them, an AHL-dependent QS system has been found to control biofilm formation in several bacterial species, including Pseudomonas aeruginosa, Aeromonas hydrophila, Burkholderia cepacia, and Serratia liquefaciens. Bacterial biofilm is a structured community of bacterial cells enclosed in a self-produced polymeric matrix that adheres to an inert or living surface. Extracellular signal molecules have been implicated in biofilm formation. Agrobacterium tumefaciens strain NT1(traR, tra::lacZ749) and Chromobacterium violaceum strain CV026 are used as biosensors to detect AHL signals. Quorum sensing in lactic acid bacteria involves peptides that are directly sensed by membrane-located histidine kinases, after which the signal is transmitted to an intracellular regulator. In the nisin autoregulation process in Lactococcus lactis, the NisK protein acts as the sensor for nisin, and NisR protein as the response regulator activatingthe transcription of target genes. For control over growth and survival in bacterial communities, various strategies need to be developed by which receptors of the signal molecules are interfered with or the synthesis and release of the molecules is controlled. However, much is still unknown about the metabolic processes involved in such signal transduction and whether or not various foods and food ingredients may affect communication between spoilage or pathogenic bacteria. In five to ten years, we will be able to discover new signal molecules, some of which may have applications in food preservation to inhibit the growth of pathogens on foods.