• Title/Summary/Keyword: meta-pathway

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Genetic Structure of xyl Gene Cluster Responsible for Complete Degradation of (4-Chloro )Benzoate from Pseudomonas sp. S-47

  • Park, Dong-Woo;Lee, Kyoung;Chae, Jong-Chan;Kudo, Toshiaki;Kim, Chi-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.483-489
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    • 2004
  • Pseudomonas sp. S-47 is a bacterium capable of degrading benzoate as well as 4-chlorobenzoate (4CBA). Benzoate and 4CBA are known to be degraded via a meta-cleavage pathway characterized by a series of enzymes encoded by xyl genes. The meta-cleavage pathway operon in Pseudomonas sp. S-47 encodes a set of enzymes which transform benzoate and 4CBA into TCA cycle intermediates via the meta-cleavage of (4-chloro )catechol to produce pyruvate and acetyl-CoA. In the current study, the meta-pathway gene cluster was cloned from the chromosomal DNA of S-47 strain to obtain pCS1, which included the degradation activities for 4CBA and catechol. The genetic organization of the operon was then examined by cloning the meta-pathway genes into a pBluescript SKII(+) vector. As such, the meta-pathway operon from Pseudomonas sp. S-47 was found to contain 13 genes in the order of xylXYZLTEGFlQKIH. The two regulatory genes, xylS and xylR, that control the expression of the meta-pathway operon, were located adjacently downstream of the meta-pathway operon. The xyl genes from strain S-47 exhibited a high nucleoside sequence homology to those from Pseudomonas putida mt-2, except for the xylJQK genes, which were more homologous to the corresponding three genes from P. stutzeri AN10. One open reading frame was found between the xylH and xylS genes, which may playa role of a transposase. Accordingly, the current results suggest that the xyl gene cluster in Pseudomonas sp. S-47 responsible for the complete degradation of benzoate was recombined with the corresponding genes from P. putida mt-2 and P. stutzeri AN10.

Isolation and Characterization of Aniline Degrading Bacteria (아닐린 생분해 균주의 분리 및 특성)

  • 조경연;하일호;배경숙;고영희
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.421-426
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    • 1988
  • Twenty-nine bacterial isolates capable of growing on aniline as n sole source of carbon and nitrogen were obtained. Ten of these isolates were identified. Nine isolates were Identified as Pseudomonas spp. and one was Acinetobacter sp.. Five strains among 29 isolates had one or several plasmids. Four of these five strains utilized aniline through meta pathway and one through ortho pathway. Pseudomonas acidovorans 4A1 which utilized aniline through meta pathway harbored a plasmid of ca. 230 kilobases shown to be involved in aniline metabolism.

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Prediction and Identification of Biochemical Pathway of Acteoside from Whole Genome Sequences of Abeliophyllum Distichum Nakai, Cultivar Ok Hwang 1ho (미선나무 품종 옥황 1호의 유전체를 활용한 Acteoside 생화학 합성과정 예측 및 확인)

  • Park, Jaeho;Xi, Hong;Han, Jiyun;Lee, Jeongmin;Kim, Yongsung;Lee, Jun-mi;Son, Janghyuk;Ahn, Joungjwa;Jang, Taewon;Choi, Jisoo;Park, Jongsun
    • Journal of Convergence for Information Technology
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    • v.10 no.3
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    • pp.76-91
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    • 2020
  • Whole genome sequence of Abeliophyllum distichum Nakai (Oleaceae) cultivar Ok Hwang 1 Ho, which is Korean endemic species, was recently sequenced to understand its characteristics. Acteoside is one of major useful compounds presenting various activities, and its several proposed biochemical pathways were reviewed and integrated to make precise biochemical pathway. Utilizing MetaPre-AITM which was developed for predicting secondary metabolites based on whole genome with the precise biochemical pathway of acteoside and the InfoBoss Pathway Database, we successfully rescued all enzymes involved in this pathway from the genome sequences, presenting that A. distichum cultivar Ok Hwang 1 Ho may produce acteoside. High-performance liquid chromatography result displayed that callus of A. distichum cultivar Ok Hwang 1 Ho contained acteoside as well as isoacteoside which may be derived from acteoside. Taken together, we successfully showed that MetaPre-AITM can predict secondary metabolite from plant whole genomes. In addition, this method will be efficient to predict secondary metabolites of many plant species because DNA can be analyzed more stability than chemical compounds.

Alcaligenes eutrophus 균주의 성장과 Poly-Beta-hydroxybutyrate 합성에 미치는 포도당과 암모늄농도의 영향

  • 이용우;유영제
    • Microbiology and Biotechnology Letters
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    • v.18 no.6
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    • pp.607-612
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    • 1990
  • The biodegradation of Aroclor 1242 was investigated by the mixed cultivation of the natural bacterial isolates and a genetically engineered microorganism (GEM). The natural strain of MS-1003 degraded the Aroclor 1242 through the ortho-cleavage pathway, while the other strains through the meta-cleavage pathway. When the MS-1003 strain was additionally inoculated into the 1 day culture of the DJ-26 strain and then cultivated for 2 days, the Aroclor was degraded up to 86% and resulted in increase of the meta-cleavage product. But in the MS-1003 culture inoculated with the DJ-26, degradation of the Aroclor was limited to the level of each pure culture. By the mixed cultivation of the DJ-26 strain together with the DJ-12 or its GEM strain of DF-10, which degrades the Aroclor through the meta-cleavage pathway, degradation of the Aroclor as well as production of the meta-cleavage compound were lower than those of each pure culture. The degradation of Aroclor 1242 by the GEM strain was not improved over the parental strain. Therefore, a form of cometaboiism of Aroclor 1242 was found in the mixed culture of the DJ-26 and MS-1003 strains which degrade the Aroclor through the different metabolic pathway, but in the mixed culture of the DJ-26 and DJ-12 strains degrading Aroclor 1242 through the same pathway, a kind of competetion for the substrate was observed.

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Analysis of Enzymes of Stenotrophomonas maltophilia LK-24 Associated with Phenol Degradation (Stenotrophomonas maltophilia LK-24의 페놀분해 관련 효소)

  • Kim, Jeong-Dong;Kang, Kook-Hee
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.37-46
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    • 2004
  • The analysis of enzymes associated with metabolism of phenolics by Stenotrophomonas maltophilia LK-24 was conducted. To identify metabolites of phenol and phenol compound, we investigated enzymes of S. maltophilia LK-24 associated with degradation of phenolics. We found that phenol hydrolase, catechol-2.3-dioxygenase, 2-hydroxymuconic semialdehyde dehydrogenase, 2-hydroxymuconic semialdehyde hydroxylase and acetaldehyde dehydrogenase were activated. The results showed that phenolics were gone through the meta-pathway ring cleavage. The results will contribute greatly to understand metabolic pathways of phenol and it is possible to make some assessment of the feasibility of using S. maltophilia LK-24 for the treatments of phenolic-contaminated waste streams.

Roles of the meta- and the ortho-Cleavage Pathways for the Efficient Utilization of Aromatic Hydrocarbons by Sphingomonas yanoikuyae Bl

  • 송정민;김영민;Gerben J. Zylstra;김응빈
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.245-245
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    • 2002
  • Catabolic pathways for the degradation of various aromatics by Sphingomonas yanoikuyae Bl are intertwined, joining at the level of substituted benzoates, which are further degraded vita ring cleavage reactions. The mutant strain EK497, which was constructed by deleting a large DNA region containing most of the genes for biphenyl, naphthalene, m-xylene, and m-toluate degradation, was unable to grow on all of the aromatics tested except for benzoate as the sole source of carbon and energy.S. yanoikuyae EK497 was found to possess only catechol ortho-ring cleavage activity due to deletion of the genes for the meta-cleavage pathway. Wild-type S. yanoikuyae Bl grown on benzoate has both catechol orthoand meta-cleavage activity. However, m-xylene and m-toluate, which are metabolized through methylbenzoate, and biphenyl, which is metabolized through benzoate, induce only the meta-cleavage pathway, suggesting the presence of a substrate-dependent induction mechanism.

Roles of the meta- and the ortho-Cleavage Pathways for the Efficient Utilization of Aromatic Hydrocarbons by Sphingomonas yanoikuyae Bl

  • Jeongmin Song;Junghee Sung;Kim, Young-Min;Gerben J. Zylstra;Kim, Eungbin
    • Journal of Microbiology
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    • v.38 no.4
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    • pp.245-249
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    • 2000
  • Catabolic pathways for the degradation of various aromatics by Sphingomonas yanoikuyae Bl are intertwined, joining at the level of substituted benzoates, which are further degraded vita ring cleavage reactions. The mutant strain EK497, which was constructed by deleting a large DNA region containing most of the genes for biphenyl, naphthalene, m-xylene, and m-toluate degradation, was unable to grow on all of the aromatics tested except for benzoate as the sole source of carbon and energy.S. yanoikuyae EK497 was found to possess only catechol ortho-ring cleavage activity due to deletion of the genes for the meta-cleavage pathway. Wild-type S. yanoikuyae Bl grown on benzoate has both catechol orthoand meta-cleavage activity. However, m-xylene and m-toluate, which are metabolized through methylbenzoate, and biphenyl, which is metabolized through benzoate, induce only the meta-cleavage pathway, suggesting the presence of a substrate-dependent induction mechanism.

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Characterization of the BTEX-degrading pathway genes in Ralstonia sp. strain PHS1

  • Lee, Sun-Bok;Lee, Sung-Kuk
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2001.06a
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    • pp.116-117
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    • 2001
  • A thermotolerant bacterium, designated as PHS1, was isolated from a hot spring in Pohang, Korea, on the basis of its ability to grow on BTEX as a sole carbon source. We cloned and sequenced the entire BTEX-degrading pathway genes of PHS1 and found that two multicomponent mono-oxygenases together with meta-pathway genes are responsible for the BTEX biodegradation.

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Induction by Carvone of the Polychlorinated Biphenyl (PCB)-Degradative Pathway in Alcaligenes eutrophus H850 and Its Molecular Monitoring

  • Park, Young-In;So, Jae-Seong;Koh, Sung-Cheol
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.804-810
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    • 1999
  • There is a possibility that carvone, a monoterpene from spearmint (Mentha spicata), could induce the bph degradative pathway and genes in Alcaligenes eutrophus H850, which is a known Gram-negative PCB degrader with a broad substrate specificity that was thoroughly investigated with Arthrobacter sp. BIB, a Gram-positive PCB degrader. The strains BIB and H850 were unable to utilize and grow on the plant terpene [(R)-(-)-carvone] (50ppm) to be recognized as a sole carbon source. Nevertheless, the carvone did induce 2,3-dihydroxybiphenyl 1,2-dioxygenase (encoded by bphC) in the strain B lB, as observed by a resting cell assay that monitors accumulation of a yellow meta ring fission product from 4,4'-dichlorobiphenyl (DCBp). The monoterpene, however, did not appear to induce the meta cleavage pathway in the strain H850. Instead, an assumption was made that the strain might be using an alternative pathway, probably the ortho-cleavage pathway. A reverse transcription (RT)-PCR system, utilizing primers designed from a conserved region of the bphC gene of Arthrobacter sp. M5, was employed to verify the occurrence of the alternative pathway. A successful amplification (182bp) of mRNA transcribed from the N-terminal region of the bphC gene was accomplished in H850 cells induced by carvone (50ppm) as well as in biphenyl-growth cells. It is, therefore, likely that H850 possesses a specific PCB degradation pathway and hence a different substrate specificity compared with B1B. This study will contribute to an elucidation of the dynamic aspects of PCB bioremediation in terms of roles played by PCB degraders and plant terpenes as natural inducer substrates that are ubiquitous and environmentally compatible.

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Catabolic Degradation of 4-Chlorobiphenyl by Pseudomonas sp. DJ-12 via Consecutive Reaction of meta-Cleavage and Hydrolytic Dechlorination

  • Chae, Jong-Chan;Kim, Eunheui;Park, Sang-Ho;Kim, Chi-Kyung
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.6
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    • pp.449-455
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    • 2000
  • Pseudomonas sp. strain DJ-12 is a bacterial isolate capable of degrading 4-chlorobiphenyl (4CBP) as a carbon and energy source. The catabolic degradation of 4CBP by the strain DJ-12 was studied along with the genetic organization of the genes responsible for the crucial steps of the catabolic degradation. The catabolic pathway was characterized as being conducted by consecutive reactions of the meta-cleavage of 4CBP, hydrolytic dechlorination of 4-chlorobenzoate (4CBA), hydroxylation of 4-hydroxybenzoate, and meta-cleavage of protocatechuate. The pcbC gene responsible for the meta-cleavage of 4CBP only showed a 30 to 40% homology in its deduced amino acid sequence compared to those of the corresponding genes from other strains. The amino acid sequence of 4CBA-CoA dechlorinase showed an 86% homology with that of Pseudomonas sp. CBS3, yet only a 50% homology with that of Arthrobacter spp. However, the fcb genes for the hydrolytic dechlorination of 4CBA in Pseudomonas sp. DJ-12 showed an uniquely different organization from those of CBS3 and other reported strains. Accordingly, these results indicate that strain DJ-12 can degrade 4CBA completely via meta-cleavage and hydrolytic dechlorination using enzymes that are uniquely different in their amino acid sequences from those of other bacterial strains with the same degradation activities.

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