• Title/Summary/Keyword: bphC

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Cloning and Phylogenetic Analysis of Two Different bphC Genes and bphD Gene From PCB-Degrading Bacterium, Pseudomonas sp. Strain SY5

  • Na, Kyung-Su;Kim, Seong-Jun;Kubo, Motoki;Chung, Seon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.11 no.4
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    • pp.668-676
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    • 2001
  • Pseudomonas sp. strain SY5 is a PCB-degrading bacterium [24] that includes two different enzymes (BphC1 and BphC2) encoding 2,3-dihdroxybiphenyl 1,2-dioxygenase and BphD encoding 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase. The bphC1 and bphC2 genes were found to consist of 897 based encoding 299 amino acids and 882 bases encoding 294 amino acids, respectively, whereas the bphD gene consisted of 861 bases encoding 287 amino acids. According to a homology search, a 50% and 39% similarity between the bphC1 and bphC2 genes at the nucleotide and amino acid level was shown, respectively. The bphC1 gene showed a 38% and 45% similarity at the amino acid level to Alcaligenes eutrophus A5 and Rhodococcus rhodochrous, respectively, whereas, bphC2 showed a 95% and 43% similarity, respectively. A comparison of the deduced amino acid sequence of the bphD product of Pseudomonas sp. SY5 with that of A. eutrophus A5, Pseudomons sp. KKS102, and LB400 showed a sequence identity of 92, 92, and 79%, respectively. Strain SY5 was originally isolated from municipal sewage containing recalcitrant organic compounds an found to have a high degradability of various aromatic compounds [23]. The current study found that strain SY5 had two extradiol-type dioxygenases, which did not hybridize with each other as they had a low similarity, yet a similar structure of evolutionarily conserved amino acids residues for catalytic activity between BphC1 and BphC2 was observed.

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Heat-Killed and Live Enterococcus faecalis Attenuates Enlarged Prostate in an Animal Model of Benign Prostatic Hyperplasia

  • Choi, Young-Jin;Fan, Meiqi;Tang, Yujiao;Iwasa, Masahiro;Han, Kwon-Il;Lee, Hongchan;Hwang, Ji-Young;Lee, Bokyung;Kim, Eun-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.31 no.8
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    • pp.1134-1143
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    • 2021
  • In the present study, we investigated the inhibitory effect of heat-killed Enterococcus faecalis (E. faecalis) and live E. faecalis on benign prostatic hyperplasia (BPH). The BPH rat model was established by administering male rats with testosterone propionate (TP, 5 mg/kg, in corn oil) via subcutaneous injections daily for four weeks after castration. The rats were divided into five groups: Con, corn oil-injected (s.c.) + DW administration; BPH, TP (5 mg/kg, s.c.) + DW administration; BPH+K_EF, TP (5 mg/kg, s.c.) + heat-killed E. faecalis (7.5 × 1012 CFU/g, 2.21 mg/kg) administration; BPH+L_EF, TP (5 mg/kg, s.c.) + live E. faecalis (1 × 1011 CFU/g, 166 mg/kg) administration; BPH+Fi, TP (5 mg/kg, s.c.) + finasteride (1 mg/kg) administration. In both of BPH+K_EF and BPH+L_EF groups, the prostate weight decreased and histological changes due to TP treatment recovered to the level of the Con group. Both of these groups also showed regulation of androgen-signaling factors, growth factors, and apoptosis-related factors in prostate tissue. E. faecalis exhibited an inhibitory effect on benign prostatic hyperplasia, and even heat-killed E. faecalis showed similar efficacy on the live cells in the BPH rat model. As the first investigation into the effect of heat-killed and live E. faecalis on BPH, our study suggests that heat-killed E. faecalis might be a food additive candidate for use in various foods, regardless of heat processing.

Biological Response of Resistant Genes to Korean Brown Planthopper, Nilaparvata lugens Stål (벼멸구 저항성 유전자에 대한 국내 벼멸구의 생물적 반응 연구)

  • Choi, Nak Jung;Kim, Gwang-Ho;Baik, Chai-Hun;Lee, Bong-Choon
    • Journal of Life Science
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    • v.29 no.2
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    • pp.202-208
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    • 2019
  • Brown planthopper (BPH), Nilaparvata lugens Stål (Hemiptera: Delphacidae), is one of the most important migratory pests damaging rice in Korea. It invades annually from tropical and subtropical areas via continental air streams. It is necessary to determine the resistance levels of rice varieties in order to control efficiency. The honeydew excretion, development, and reproduction of the migratory BPH were studied by region in a laboratory at $25{\pm}2^{\circ}C$ and $65{\pm}5%\;RH$ and a 16L: 8D photoperiodism conducted on three BPH resistant genes: Bph1, Bph2, and Bph18. The information obtained was reported using the jackknife method, and we created life table statistics accordingly. The feeding amount of Bph1 resistant gene was lower than that of resistant genes. The developmental periods of immature stages ranged from $13.7{\pm}0.10d$ on Bph2 (Namhae, 2015) to $18.5{\pm}1.06d$ on Bph2 (Sacheon, 2016). Reproductive period and female longevity were longest on the non-resistant genes, Bph2 and Bph18 (except 1980s), and the highest fecundity of N. lugens was observed on the two BPH resistant genes. Highest net reproductive rates ($R_0$) were calculated on Bph2 by region. Intrinsic rates of population increase ($r_m$) showed a difference in resistant genes by region. These population parameters showed that migratory regions and biological characteristics of N. lugens vary annually.

Molecular Mapping of Resistant Genes to Brown Planthopper, Bphl and bph2, in Rice

  • Cha, Young-Soon;Cho, Yong-Gu;Shin, Kyeong-Og;Yeo, Un-Sang;Choi, Jae-Eul;Eun, Moo-Young
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.4
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    • pp.345-349
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    • 1999
  • This study was carried out to map Bphl and bph2 gene in Mudgo and Sangju13 (Oryza sativa L.) respectively conferring resistance to brown plan-thopper (BPH) and to establish the marker-assisted selection (MAS) system. Bulked seedling (grown for 20 days) test was conducted with the 73 F4 lines derived from a cross between Nagdongbyeo and Mudgo for Bphl and with 53 BC3F5 lines derived from the Milyang95/Sangju13 cross for bph2. Bph1 was mapped between RG413 and RG901 on chromo-some 12 at a distance of 7.5 cM from RG413 and 8.4 cM from RG90l. A recessive gene bph2 was located near RZ76 on chromosome 12 at a distance of 14.4 cM. Bphl and bph2 were linked to each other with a distance of about 30 cM. An RFLP marker, RG413 linked to Bphl, was converted to an STS marker to facilitate the marker-assisted selection. BPH resistant genotypes could be selected with 92% accuracy in a population derived from a line of NM47-B-B.

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Production of Bacteriophytochrome Specific Antibodies of Deinococcus radiodurans (Deinococcus radiodurans 박테리오피토크롬 특이 항체들의 생산)

  • Kim, Tae-Lim;Hahn, Tae-Ryong;Bhoo, Seong-Hee
    • Journal of Applied Biological Chemistry
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    • v.53 no.2
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    • pp.112-115
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    • 2010
  • To analyze the surface properties of bacteriophytochrome (BphP), five (2B8, 2C11, 3B2, 3D2, 3H7) anti-BphP monoclonal antibodies were produced by using full-length of BphP of Deinococcus radiodurnas. 2B8 and 2C11 preferentially recognized the epitopes at N-terminal region of BphP, whereas 3B2, 3D2 and 3H7 showed preferential affinities to the epitopes of C-terminal region of BphP.

Monitoring Expression of bphC Gene from Ralstonia eutropha H85O Induced by Plant Terpenes in Soil

  • Jung, Kyung-Ja;Kim, Byung-Hyuk;Kim, Eungbin;So, Jae-Seong;Koh, Sung-Cheol
    • Journal of Microbiology
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    • v.40 no.4
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    • pp.340-343
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    • 2002
  • A PCB degrader, Ralstonia eutropha H850 was shown to induce bphC gene encoding 2,3-dihydroxy-biphenyl-1,2-dioxygenase in a carvone-amended pure culture in our previous study (Park et al.,1999). The present study was carried out to examine how plant terpenes, as natural substrates, would cause an expression of a PCB degradative gene in soil that was amended with terpenes. The population of Ralstonia eutropha H850 was maintained at least around 10$\^$8/ (CFU/g fresh soil) in the soil amended with carvone or limonene in the presence of succinate as a growth substrate at 50 th day. The gene expression was monitored by RT-PCR using total RNA directly extracted from each soil and bphC gene primers. The bphC gene expression of the seeded strain H850 was observed in the soil amended with biphenyl (4 days) but not with succinate, carvone and limonene. These results indicate that terpenes widely distributed in nature could be a potential inducing substrate for effective PCB biodegration in the soil but their bioavailability and specific induction behavior should be taken into account before PCB bioremediation implementation.

Fine Mapping of the Rice Bph1 Gene, which Confers Resistance to the Brown Planthopper (Nilaparvata lugens Stal), and Development of STS Markers for Marker-assisted Selection

  • Cha, Young-Soon;Ji, Hyeonso;Yun, Doh-Won;Ahn, Byoung-Ohg;Lee, Myung Chul;Suh, Seok-Cheol;Lee, Chun Seok;Ahn, Eok Keun;Jeon, Yong-Hee;Jin, Il-Doo;Sohn, Jae-Keun;Koh, Hee-Jong;Eun, Moo-Young
    • Molecules and Cells
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    • v.26 no.2
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    • pp.146-151
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    • 2008
  • The brown planthopper (BPH) is a major insect pest in rice, and damages these plants by sucking phloem-sap and transmitting viral diseases. Many BPH resistance genes have been identified in indica varieties and wild rice accessions, but none has yet been cloned. In the present study we report fine mapping of the region containing the Bph1 locus, which enabled us to perform marker-aided selection (MAS). We used 273 F8 recombinant inbred lines (RILs) derived from a cross between Cheongcheongbyeo, an indica type variety harboring Bph1 from Mudgo, and Hwayeongbyeo, a BPH susceptible japonica variety. By random amplification of polymorphic DNA (RAPD) analysis using 656 random 10-mer primers, three RAPD markers (OPH09, OPA10 and OPA15) linked to Bph1 were identified and converted to SCAR (sequence characterized amplified region) markers. These markers were found to be contained in two BAC clones derived from chromosome 12: OPH09 on OSJNBa0011B18, and both OPA10 and OPA15 on OSJNBa0040E10. By sequence analysis of ten additional BAC clones evenly distributed between OSJNBa0011B18 and OSJNBa0040E10, we developed 15 STS markers. Of these, pBPH4 and pBPH14 flanked Bph1 at distances of 0.2 cM and 0.8 cM, respectively. The STS markers pBPH9, pBPH19, pBPH20, and pBPH21 co-segregated with Bph1. These markers were shown to be very useful for marker-assisted selection (MAS) in breeding populations of 32 F6 RILs from a cross between Andabyeo and IR71190, and 32 F5 RILs from a cross between Andabyeo and Suwon452.

Identification of the bphC Gene for meta-Cleavage of Aromatic Pollutants from a Metagenomic Library Derived from Lake Waters

  • Moon Mi-Sook;Lee Dong-Hun;Kim Chi-Kyung
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.5
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    • pp.393-399
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    • 2004
  • Useful genes can be Screened from various environments by construction of metagenomic DNA libraries. In this study, water samples were collected from several lakes in mid Korea, and analyzed by T-RFLP to examine diversities of the microbial communities. The crude DNAs r were extracted by the SDS-based freezing-thawing method, and then further purified using an $UltraClean^{TM}$ kit (MoBio, USA). The metagenomic libraries were constructed with the DNAs partially digested with EcoR I, BamH I, and Sac II in Escherichia coli DH 10B using the pBACe3.6 vector. About 44.0 Mb of metagenomic libraries were obtained with average inserts 13-15 kb in size. The bphC genes responsible for degradation of aromatic hydrocarbons via mets-cleavage were identified from the metagenomic libraries by colony hybridization using the bphC specific sequence as a probe. The 2,3-dihydroxybiphenyl (2, 3-DHBP) dioxygenase gene (bphC ), capable of degradation of 2,3-DHBP, was cloned and its nucleotide Sequences analyzed. The genes consisted of 966 and 897 base pairs with an ATG initiation codon and a TGA termination codon. The activity of the 2,3-DHBP dioxygenase was highly expressed to 2,3-DHBP and Showed a broad substrate range to 2,3-DHBP, catechol, 3-methylcatechol and 4-methylcatechol. These results in-dicated that the bphC gene identified from the metagenomes derived from lake water might be useful in the development of a potent strain for degradation of aromatic pollutants.

The Predatory Behavior of Green Mirid Bug, Cytorhinus lividipennis Reuter, on Brown Planthopper Eggs in Different Temperature Conditions (온도에 따른 등검은황록장님노린재 (Cyrtorhinus lividipennis Reuter)의 벼멸구 알에 대한 포식습성)

  • ;;;K.L. Heong
    • Korean journal of applied entomology
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    • v.34 no.3
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    • pp.234-242
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    • 1995
  • The functional responses of the female Cyrtorhinus lividipennis on brown planthopper (BPH), Nilaparvata lugens, eggs and their predation behavior were investigated at six temperature conditions; 20, 23, 26, 29, 32, and $35^{\circ}C$. C. lividipennis was found to prefer young BPH eggs, especially 3-day-old eggs the most. The functional responses for female C. lividipennis on BPH eggs fitted the Holling's Type-I true predator-prey Rogers' (1972) model at all temperatures tested except $35^{\circ}C$ at which the negative Th value was produced. With the temperature increased up to 32$^{\circ}C$, the instantaneous attacking rate(a) increased from 0.1923 at $20^{\circ}C$ to 0.5085 at $32^{\circ}C$, while the handling time (Th) was gradually decreased as low as 0.0151 at $32^{\circ}C$. C. lividipenis preferred the BPH eggs laid on the upper part of rice stem when the BPH egg density was high, but there was no significant difference in the preference when the egg density was low. The preference was more obvious in high temperature conditions such as above 29$^{\circ}C$.

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Versatile Catabolic Properties of Tn4371-encoded bph Pathway in Comamonas testosteroni (Formerly Pseudomonas sp.) NCIMB 10643

  • Kim, Jong-Soo;Kim, Ji-Hyun;Ryu, Eun-Kyeong;Kim, Jin-Kyoo;Kim, Chi-Kyung;Hwang, In-Gyu;Lee, Kyoung
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.302-311
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    • 2004
  • Comamonas testosteroni (formerly Pseudomonas sp.) NCIMB 10643 can grow on biphenyl and alkylbenzenes $(C_2-C_7)$ via 3-substituted catechols. Thus, to identify the genes encoding the degradation, transposon-mutagenesis was carried out using pAG408, a promoter-probe mini-transposon with a green fluorescent protein (GFP), as a reporter. A mutant, NT-1, which was unable to grow on alkylbenzenes and biphenyl, accumulated catechols and exhibited an enhanced expression of GFP upon exposure to these substrates, indicating that the gfp had been inserted in a gene encoding a broad substrate range catechol 2,3-dioxygenase. The genes (2,826 bp) flanking the gfp cloned from an SphI-digested fragment contained three complete open reading frames that were designated bphCDorfl. The deduced amino acid sequences of bphCDorfl were identical to 2,3-dihydroxybiphenyl 1,2-dioxygenase (BphC), 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase (BphD), and OrfI, respectively, that are all involved in the degradation of biphenyl/4-chlorobiphenyl (bph) by Ralstonia oxalatica A5. The deduced amino acid sequence of the orfl revealed a similarity to those of outer membrane proteins belonging to the OmpW family. The introduction of the bphCDorfl genes enabled the NT-l mutant to grow on aromatic hydrocarbons. In addition, PCR analysis indicated that the DNA sequence and gene organization of the bph operon were closely related to those in the bph operon from Tn4371 identified in strain A5. Furthermore, strain A5 was also able to grow on a similar set of alkylbenzenes as strain NCIMB 10643, demonstrating that, among the identified aromatic hydrocarbon degradation pathways, the bph degradation pathway related to Tn4371 was the most versatile in catabolizing a variety of aromatic hydrocarbons of mono- and bicyclic benzenes.