• Title/Summary/Keyword: Photobacterium

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Expression of the Genes Involved in the Synthesis of Riboflavin from Photobacterium species of Bioluminescent Marine Bacteria (해양 발광 박테리아 Photobacterium Species의 Riboflavin 생합성에 관여하는 유전자들의 발현)

  • 이찬용
    • Korean Journal of Microbiology
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    • v.36 no.1
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    • pp.1-7
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    • 2000
  • The genes involved in riboflavin synthesis (ribI, II, III, and IV) were found immediately downstream of luxG in the lux operon from Photobacterium species. The single stranded DNA containing the intergenic region of lux genes and rib genes from Photobacterium phosphoreum was fully protected by P. phosphoreum mRNA from the S1 nuclease mapping assay suggesting that a transcriptional terminator was not present in the region. In addition, the levels of riboflavin synthase activity in P. phosphoreum was increased during the development of bacterial bioluminescence in the same fashion as the luciferase and fatty acid reductase activities. Insertion of the Photobacterium leiognathi DNA extending from luxB to ribII, between a strong lux promoter and a reporter gene (chloramphenicol acetyltransferase, CAT) and transferred by conjugation into P. leiognathi, did not affect expression of reporter gene. Moreover the CAT gene was not expressed in an analogous construct missing the lux promoter indicating that a promoter was not present in this region. Based on the data here, it can be concluded that the lux genes and rib genes in Photobacterium species are under common regulation.

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Isolation and Identification of Urease-Positive Photobacterium sp. Strain HA-2 from Sea Water (해수에서 urease 양성 Photobacterium sp. Strain HA-2의 분리 및 동정)

  • Kim, Kang-Jin;No, A-Reum;Park, Kwon-Sam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.42 no.6
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    • pp.531-536
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    • 2009
  • A urease-positive bacterium isolated from sea water was identified as Photobacterium sp. by morphological, biochemical, and 16s rRNA gene analyses and named Photobacterium sp. strain HA-2. 2.0-fold increase enzyme activity was observed in LB medium containing 3% NaCl and 0.1% urea or not and the enzyme activity was 16.0-fold lower compared to urease-positive Vibrio parahaemolyticus AQ4673 strain when grown in the LB medium containing 3% NaCl with 0.1% urea. The cloning and sequencing of Photobacterium sp. strain HA-2 urease gene cluster is currently being analyzed in our laboratory.

Genetic Characterization of the Urease Gene Cluster in Photobacterium sp. Strain HA-2 Isolated from Seawater (해수에서 분리한 Photobacterium sp. Strain HA-2가 보유하는 요소분해효소 유전자의 유전적 특징)

  • Kim, Tae Ok;Park, Kwon Sam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.5
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    • pp.639-643
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    • 2015
  • In this study, we cloned and sequenced the 15,204-bp DNA region containing the gene cluster for urease production from the chromosome of the environmental Photobacterium sp. strain HA-2. We identified 15 open reading frames (ORFs) and the G+C content was 40.3%. The urease gene cluster of Photobacterium sp. strain HA-2 consisted of seven genes, namely, ureDABCEF and ureG. There were five ORFs of urease genes in the opposite direction, which were homologous to the nickel transport operons (nik) of Vibrio parahaemolyticus and Escherichia coli. The genetic organization and sequences of the urease genes of Photobacterium sp. strain HA-2 resembled those found in Vibrio fischeri and V. parahaemolyticus.

Characterization of Photobacterium sp. YW2207 isolated from rainbow trout (Oncorhynchus mykiss) raised in a fresh water farm in South Korea (국내 양식 무지개송어(Oncorhynchus mykiss)에서 분리된 Photobacterium sp. YW2207의 특성)

  • Hyunwoo Kim;Eunsup Lee;Sung Jun Lee;Haneul Kim;So-Ra Han;Tae-Jin Oh;Myoung Sug Kim;Soo-Jin Kim;Se Ryun Kwon
    • Journal of fish pathology
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    • v.36 no.2
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    • pp.251-261
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    • 2023
  • Photobacterium sp. YW2207 was isolated from rainbow trout raised in a fish farm located in Yeongwol-gun, Gangwon Province, South Korea. Based on 16S rRNA sequence analysis and phylogenetic analysis, it was confirmed that Photobacterium sp. YW2207 showed 100% similarity with Photobacterium piscicola and Photobacterium phosphoreum, and 94.6% similarity with P. damselae subsp. damselae. Biochemical analysis revealed that Photobacterium sp. YW2207 is a Gram-negative, motile bacterium with a cell size of 1.5~3×3~5 ㎛. The bacteria were cultured on nutrient agar, brain heart infusion agar, Muller-Hinton agar, tryptic soy agar, and thiosulfate citrate bile sucrose agar with NaCl concentrations ranging from 0 to 2.5%. The API50CHE and API20E tests indicated lower utilization capabilities compared to the P. damselae strains provided in the API database. Furthermore, unlike most Photobacterium species, Photobacterium sp. YW2207 presented negative for catalase test. Results from the flow cytometric measurement indicated that Photobacterium sp. YW2207 exhibited a more diverse distribution of cell sizes and had larger cell sizes compared with P. damselae subsp. damselae. Minimum inhibitory concentration tests showed that Photobacterium sp. YW2207 had low susceptibility to β-Lactam and aminoglycoside antibiotics, while having high susceptibility to tetracycline, doxycycline, and quinolone antibiotics. Pathogenicity on rainbow trout revealed that an immersion of 1×105 CFU/ml did not cause mortality or clinical symptoms.

Characterization of Photobacterium damselae subsp. piscicida isolated from cultured starry flounder, Platichthys stellatus in Korea (우리나라 양식 강도다리, Platichthys stellatus에서 분리된 Photobacterium damselae subsp. piscicida의 특성)

  • Cho, Young Ah;Han, Hyun-Ja;Mun, Hee Eun;Jung, Sung Hee;Park, Myoung Ae;Kim, Jin Woo
    • Journal of fish pathology
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    • v.26 no.2
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    • pp.77-88
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    • 2013
  • Starry flounder, Platichthys stellatus (body length $4.4{\pm}0.51cm$) that became sick during an outbreak of disease at mariculture facilities at Ulsan, Korea in August of 2012, were examined to identify the cause of the disease. Diseased fish didn't show a unique sign, but the oxidase-positive and gram negative rod was isolated from moribund fish. The bacterium was revealed as Photobacterium damselae subsp. piscicida by biochemical analysis and sequence analysis of the 16S rRNA and capsular polysaccharide (CPS) genes. The isolates (AD5) was carrying susceptible to ofloxacin and gentamycin and showed high growth value at $18^{\circ}C$ and $25^{\circ}C$ compared to four other P. damsela strains.

Studies on the Culture Media and the Optimal Storage Conditions of Bioluminescent Bacteria Photobacterium phosphoreum (생체발광균주 Photobacterium phosphoreum의 배양배지 및 최적 저장조건에 관한 연구)

  • 조동욱;전억한;김병용;김은기;함영태
    • Korean Journal of Microbiology
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    • v.36 no.1
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    • pp.74-78
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    • 2000
  • Vibrio, Photobacterium, Alteromonas and Xenorhabdus species are capable of emitting light, called bioluminescence. They exist in marine, freshwater and terrestrial environments. Bacterial bioluminescent reaction is that reduced riboflavin phosphates and a long-chain aldehyde are oxidized in the presence of molecular oxygen and enzyme luciferase. This experiment aims to develop the proper culture media and to optimize the storage condition for the recovery of bioluminescent activity in Photobacterium phosphoreum. The Luria broth (LB) medium was modified for cultivation of Photobacterium phophoreum, called as modified LB(mLB) medium. The mLB medium is LB fortified with 3% glycerol and 1.5% NaCl. In mLB medium. bacterial growth and bioluminescent activity are 25% higher than those in a Nutrient broth medium. When the cell stocks were stored at $-20^{\circ}C$, $-70^{\circ}C$ and LN2 for 3 months, cell growth and bioluminescent activity of culture after stored at $-20^{\circ}C$ were better than those of other treatments. The highest bioluminescent activity obtained at the late exponential phase in all treatments. When the cell stock was freeze-dried with 5% adonitol as a cryoprotectant, the recovery of cell was better than those of control and freeze-dried cell stock without addition of cryoprotectant.

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The Functions of the Riboflavin Genes in the lux Operon from Photobacterium Species (Photobacterium Species의 lux 오페론에서 발견된 Riboflavin 생합성 유전자들의 기능)

  • 이찬용;임종호
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.173-179
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    • 2002
  • The functions of riboflavin synthesis genes ( ribI,II,III and IV) found immediately downstream of luxG in the lux operon from Photobacterium species were identified using the biochemical and genetical analysis. The ribI-III gene codes for protein corresponding to that coded by the second (riboflavin synthase), third (3,4-dihydroxy 2-butanone 4-phosphate synthase/GTP cyclohydrolase II) and fourth (lumazine synthase) gene, respectively, of Bacillus subtilis rib operon with the respective gene procuct sharing 41-50% amino acid sequence identity. Unexpectedly, the sequence of the ribIV product of Photobacterium phosphoreum does not correspond in sequence to the protein encoded by the fifth rib gene of Bacillus subtilis. Instead the gene (ribIV) codes for a polypeptide similar in sequence to GTP cyclohydrolase II of Escherichia coli and the carboxy terminal domain of the third rib gene from Bacillus subtilis. Complementation of Escherichia coli riboflavin auxotrophs showed that the function of the gene products of ribII and ribIV are DHBP synthase and GTP cyclohydrolase II, respectively. In addition the experiment, showing that increase in thermal stability of riboflavin synthase coded by ribIon coexpression with ribIII, provided indirect evidence that the latter gene codes for lumazine synthase.

Generation and Expression of Amino-Terminal Domain of the Gene Coding for the Lumazine Protein from Photobacterium phosphoreum (발광 박테리아 Photobacterium phosphoreum의 Lumazine Protein을 코드 하는 유전자의 염기 서열 분석 및 발현)

  • Woo Young-Eun;Kim So-Young;Lee Chan-Yong
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.306-311
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    • 2005
  • In this study, the amino-terminal half truncated lump and the whole lump genes from Photobacterium phosphoreum coding for the lumazine protein were cloned by polymerase chain reaction and expressed in Escherichia coli. To identifiy of the binding site of the ligand or substrate, the amino acid identities from the sequences of the lumazine protein, yellow fluorescent protein, and riboflavin synthase from different organisms were also compared and analyzed.

The Lux Genes and Riboflavin Genes in Bioluminescent System of Photobacterium leiognathi Are under Common Regulation

  • Sung, Nack-Do;Lee, Chan-Yong
    • Journal of Photoscience
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    • v.11 no.1
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    • pp.41-45
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    • 2004
  • The key riboflavin synthesis genes are located immediately downstream of luxG in the lux operon from Photobacterium leiognathi. It is of interest that a site capable of forming a rho-independent terminator does not appear to be present between luxG and ribE in our previous data. These results raise the question of whether the transcription of lux and rib genes is integrated or not. In order to answer the question, in vivo transcriptional assay and Southern blot were examined. These studies demonstrate that neither transcriptional terminator nor promoter site is present in the intergenic region between of lux and rib genes as well as that the riboflavin genes are single copy in a chromosome of Photobacterium leiognathi.

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Studies on the Response of Photobacterium phosphoreum to the Volatile Substances (휘발성 물질에 대한 Photobacterium phosphoreum의 Bioluminescence의 변화)

  • 이은수;이용제;김은기;이정건;전억한
    • Microbiology and Biotechnology Letters
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    • v.27 no.6
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    • pp.484-490
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    • 1999
  • Various materials including sodium alginate, k-carrageenan, collagen and polyacrylamide were studied in order to maintain the stability of bioluminescence of Photobacterium for the monitoring of volatile toxic substances. Kinetic parameters of specific rate($\mu$), and gamma(${\gamma}$) value were determined for the relationship between bioluminescence of immobilized P. phosphoreum and toxic substances. The bioluminescence intensity was found to be proportional to the concentration of toxic substances and the free cells were shown to be more sensitive than immobilized cells when volatile substances were exposed to the cells. Bioluminescence increased slightly after several minutes, which was due to the volatility of toxic compounds. Furthermore, P. phosphoreum immobilized on strontium alginate was better than cells immobilized on sodium alginate for the response to substances used.

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