Degradation of phenanthrene and naphthalene by aBurkholderiaspecies strain

2003 ◽  
Vol 49 (2) ◽  
pp. 139-144 ◽  
Author(s):  
H Kang ◽  
S Y Hwang ◽  
Y M Kim ◽  
E Kim ◽  
Y -S Kim ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Energy Source ◽  
Enzyme Assay ◽  
Gas Chromatography Mass Spectrometry ◽  
Ring Cleavage ◽  
Growth Substrate ◽  
Degrading Bacteria ◽  
Subsequent Degradation ◽  
Cleavage Enzyme ◽  
Uv Visible

Burkholderia sp. TNFYE-5 was isolated from soil for the ability to grow on phenanthrene as sole carbon and energy source. Unlike most other phenanthrene-degrading bacteria, TNFYE-5 was unable to grow on naphthalene. Growth substrate range experiments coupled with the ring-cleavage enzyme assay data suggest that TNFYE-5 initially metabolizes phenanthrene to 1-hydroxy-2-naphthoate with subsequent degradation through the phthalate and protocatechuate and β-ketoadipate pathway. A metabolite in the degradation of naphthalene by TNFYE-5 was isolated by high-pressure liquid chromatography (HPLC) and was identified as salicylate by UV-visible spectral and gas chromatography – mass spectrometry analyses. Thus, the inability to degrade salicylate is apparently one major reason for the incapability of TNFYE-5 to grow on naphthalene.Key words: Burkholderia, phenanthrene, naphthalene, phthalate, protocatechuate.

Growth of Frankia in the rhizosphere of Betula pendula, a nonhost tree species

1990 ◽  
Vol 36 (9) ◽  
pp. 649-656 ◽  
Author(s):  
A. Smolander ◽  
R. Rönkkö ◽  
E.-L. Nurmiaho-Lassila ◽  
K. Haahtela
Keyword(s):  
Electron Microscopy ◽  
Mass Spectrometry ◽  
Energy Source ◽  
Betula Pendula ◽  
Defined Medium ◽  
Gas Chromatography Mass Spectrometry ◽  
Favourable Effect ◽  
Culture Solution ◽  
Alnus Incana ◽  
Scanning Electron

The growth of Frankia in the rhizosphere of Betula pendula was studied to understand the favourable effect of this nonhost species on Alnus-nodulating Frankia in soil. Axenic Betula pendula and Alnus incana seedlings were inoculated with Frankia strains. The development of Frankia in the rhizosphere was examined by light and scanning electron microscopy. Some Frankia strains grew in the rhizosphere of Betula pendula seedlings without addition of a carbon and energy source. In addition to hyphae and sporangia, vesicles were also formed. Growth (if any) was more local and less abundant in the rhizosphere of Alnus incana seedlings. Frankia strains were grown in defined medium supplemented with tryptophan. Indole-3-ethanol was detected in the culture solution by gas chromatography – mass spectrometry. Key words: Alnus incana, Betula pendula, Frankia, indole-3-ethanol.

scholarly journals Co-production of polyhydroxybutyrate (PHB) and coenzyme Q10 (CoQ10) via no-sugar fermentation—a case by Methylobacterium sp. XJLW

2021 ◽  
Vol 71 (1) ◽  
Author(s):  
Peiwu Cui ◽  
Yunhai Shao ◽  
Yanxin Wang ◽  
Rui Zhao ◽  
Huihui Zhan ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Energy Source ◽  
Fermentation Process ◽  
Genetically Modify ◽  
Tween 80 ◽  
Batch Process ◽  
Gas Chromatography Mass Spectrometry ◽  
Fed Batch ◽  
Enhanced Production ◽  
Chromatography Mass Spectrometry

Abstract Purpose To explore a competitive PHB-producing fermentation process, this study evaluated the potential for Methylobacterium sp. XJLW to produce simultaneously PHB and coenzyme Q10 (CoQ10) using methanol as sole carbon and energy source. Methods The metabolic pathways of PHB and CoQ10 biosynthesis in Methylobacterium sp. XJLW were first mined based on the genomic and comparative transcriptomics information. Then, real-time fluorescence quantitative PCR (RT-qPCR) was employed for comparing the expression level of important genes involved in PHB and CoQ10 synthesis pathways’ response to methanol and glucose. Transmission electron microscope (TEM), gas chromatography/mass spectrometry (GC-MS), nuclear magnetic resonance (NMR), Fourier transformation infrared spectrum (FT-IR), and liquid chromatography/mass spectrometry (LC-MS) methods were used to elucidate the yield and structure of PHB and CoQ10, respectively. PHB and CoQ10 productivity of Methylobacterium sp. XJLW were evaluated in Erlenmeyer flask for medium optimization, and in a 5-L bioreactor for methanol fed-batch strategy according to dissolved oxygen (DO) and pH control. Results Comparative genomics analysis showed that the PHB and CoQ10 biosynthesis pathways co-exist in Methylobacterium sp. XJLW. Transcriptomics analysis showed that the transcription level of key genes in both pathways responding to methanol was significantly higher than that responding to glucose. Correspondingly, strain Methylobacterium sp. XJLW can produce PHB and CoQ10 simultaneously with higher yield using cheap and abundant methanol than using glucose as sole carbon and energy source. The isolated products showed the structure characteristics same to that of standard PHB and CoQ10. The optimal medium and cultural conditions for PHB and CoQ10 co-production by Methylobacterium sp. XJLW was in M3 medium containing 7.918 g L-1 methanol, 0.5 g L-1 of ammonium sulfate, 0.1% (v/v) of Tween 80, and 1.0 g L-1 of sodium chloride, under 30 °C and pH 7.0. In a 5-L bioreactor coupled with methanol fed-batch process, a maximum DCW value (46.31 g L-1) with the highest yields of PHB and CoQ10, reaching 6.94 g L-1 and 22.28 mg L-1, respectively. Conclusion Methylobacterium sp. XJLW is potential for efficiently co-producing PHB and CoQ10 employing methanol as sole carbon and energy source. However, it is still necessary to further optimize fermentation process, and genetically modify strain pathway, for enhanced production of PHB and CoQ10 simultaneously by Methylobacterium sp. XJLW. It also suggests a potential strategy to develop efficiently co-producing other high-value metabolites using methanol-based bioprocess.

scholarly journals Co-production of biopolymers and quinone via no-sugar fermentation--a case by Methylobacterium sp. XJLM

2021 ◽  
Author(s):  
Peiwu Cui ◽  
Yunhai Shao ◽  
Yanxin Wang ◽  
Rui Zhao ◽  
Huihui Zhan ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Energy Source ◽  
Fermentation Process ◽  
Genetically Modify ◽  
Tween 80 ◽  
Batch Process ◽  
Gas Chromatography Mass Spectrometry ◽  
Fed Batch ◽  
Enhanced Production ◽  
Chromatography Mass Spectrometry

Abstract Purpose To explore a competitive PHB producing fermentation process, this study evaluated the potential for Methylobacterium sp. XJLW to produce simultaneously PHB and coenzyme Q 10 (CoQ 10 ) using cheap and abundant methanol as sole carbon and energy source. Methods The metabolic pathways of PHB and CoQ 10 biosynthesis in XJLW strain were first mined based on the genomic and comparative transcriptomics information. Then, Real-time fluorescence quantitative PCR (RT-qPCR) was employed for comparing the expression level of important genes involved in PHB and CoQ10 synthesis pathways response to methanol and glucose. Transmission electron microscope (TEM), gas chromatography/mass spectrometry (GC-MS), nuclear magnetic resonance (NMR), Fourier transformation infrared spectrum (FT-IR), and liquid chromatography/mass spectrometry (LC-MS) methods, were used to elucidate the yield and structure of PHB and CoQ 10 , respectively. PHB and CoQ 10 productivity of XJLW strain were evaluated in flasks for medium optimization, and in a 5-L bioreactor for methanol fed-batch strategy according to dissolved oxygen (DO) and pH control. Results Comparative genomics and transcriptomics analysis showed that the PHB and CoQ 10 biosynthesis pathways coexist in XJLW strain, and the transcription level of key genes in both pathways response to methanol was significantly higher than that response to glucose. Correspondingly, strain XJLW can produce PHB and CoQ 10 simultaneously with higher yield using cheap and abundant methanol than using glucose as sole carbon and energy source. The isolated products showed the structure characteristics same to that of standard PHB and CoQ 10 . The optimal medium and cultural conditions for PHB and CoQ 10 co-production by XJLW strain was in M3 medium containing 1% (v/v) of methanol, 0.5 g/L of ammonium sulfate, 0.1% (v/v) of Tween 80, and 1.0 g/L of sodium chloride, under 30°C and pH 7.0. In a 5-L bioreactor coupled with methanol fed-batch process, a maximum DCW value (46.31 g/L) with the highest yields of PHB and CoQ 10 , reaching 6.94 g/L and 22.28 mg/L, respectively. Conclusion Methylobacterium sp. XJLW is potential for efficiently co-producing PHB and CoQ 10 employing methanol as sole carbon and energy source. However, it is still necessary to further optimize fermentation process, and genetically modify strain pathway, for enhanced production of PHB and CoQ 10 simultaneously by XJLW. It also suggests a potential strategy to develop efficiently co-producing other high value metabolites using methanol-based bio-process.

scholarly journals CARACTERIZACIÓN QUÍMICA Y FUNCIONAL DE METABOLITOS ANTIMICROBIANOS AISLADOS DE ASCIDIAN RHOPALAEA BIRKELANDI

2011 ◽  
Vol 3 ◽  
pp. 111
Author(s):  
Maribel Cordero ◽  
Henry Borbón ◽  
Félix R. Román ◽  
Luis Morell ◽  
Rigoberto Víquez ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Costa Rica ◽  
Magnetic Resonance ◽  
13C Nmr ◽  
Solid Phase ◽  
Proton Nuclear Magnetic Resonance ◽  
Gas Chromatography Mass Spectrometry ◽  
Spectroscopic Techniques ◽  
Ft Ir ◽  
Uv Visible

Marine ecosystems have a very large diversity of resources, most of them still partially unknown, and a few others exploited for development of new industrial and toxicological products. The objective of this investigation was to determine whether the acetone extract of the ascidia R. birkelandi from the Pacific coast of Costa Rica showed qualitative antimicrobial activity against the S. aureus bacteria and the G. candidum fungus, and to verify their main secondary metabolites in the active extract using chromatographic and spectroscopic techniques. Ascidians were collected at Tambor, Guanacaste, Costa Rica, between December 2007 and March 2008. Activity against the Gram positive bacteria and fungi was evaluated using ethanolic (95%) and acetonic extracts. Both extracts showed activity against G. candidum; however, only the acetonic extract showed activity against S. aureus. A coumarin and a hydroxyanthraquinone were isolated from a crude extract of R. birkelandi as metabolites present in the active fraction. Purification and isolation were performed by chromatographic techniques and solid phase extraction. Structural information was obtained by spectroscopic analyses: Ultraviolet (UV-Visible), Fourier Transform Infrared (FT-IR), Liquid Chromatography Tandem Mass Spectrometry (LC/MS/MS), Gas Chromatography Mass Spectrometry (GC/MS), Proton Nuclear Magnetic Resonance (1HNMR), and Carbon-13 Magnetic Resonance (13C-NMR). Further studies are recommended for characterization and quantification of the active components of this extract and the possible elucidation of the mechanisms of action. Los ecosistemas marinos poseen una gran cantidad de recursos, muchos de ellos todavía desconocidos y muy pocos de ellos explotados para el desarrollo de nuevos productos industriales y toxicológicos. Los objetivos de esta investigación fueron determinar si el extracto acetónico de la ascidia R. birkelandi de la costa pacífica de Costa Rica presenta actividad antimicrobial cualitativa contra la bacteria S. aureus y el hongo G. candidum, y determinar por técnicas cromatográficas y espectroscópicas los metabolitos secundarios principales presentes en el extracto activo. Se recolectó la ascidia en la zona de Tambor en la provincia de Guanacaste, Costa Rica, entre los meses de diciembre del 2007 y marzo del 2008. Se realizaron extractos crudos en acetona y etanol al 95% para evaluar la actividad contra una bacteria Gram positiva y un hongo. Ambos extractos presentaron actividad contra G. candidum, pero solo el extracto acetónico mostró actividad contra S. aureus. Se logró aislar y caracterizar una cumarina y una hidroxiantraquinona del extracto crudo de R. birkelandi como metabolitos presentes en la fracción responsable de la actividad biológica presentada. La purificación y el aislamiento fueron llevados a cabo con técnicas cromatográficas y de extracción en fase sólida. La información estructural fue obtenida por análisis espectroscópicos: Ultravioleta (UV-Visible), Infrarrojo con Transformada de Fourier (FT-IR), Cromatografía Líquida acoplada a Espectrometría de Masas (LC/MS/MS), Cromatografía de Gases acoplada a Masas (GC-MS), Resonancia Magnética Nuclear de Protón (1H-NMR) y Resonancia Magnética de Carbono 13 (13C-NMR). Se recomiendan estudios futuros para la caracterización y la cuantificación de los principios activos de este extracto y una posible elucidación de sus mecanismos de acción.

scholarly journals The Metabolic Pathway of 4-Aminophenol in Burkholderia sp. Strain AK-5 Differs from That of Aniline and Aniline with C-4 Substituents

2003 ◽  
Vol 69 (9) ◽  
pp. 5410-5413 ◽  
Author(s):  
Shinji Takenaka ◽  
Susumu Okugawa ◽  
Maho Kadowaki ◽  
Shuichiro Murakami ◽  
Kenji Aoki
Keyword(s):  
Mass Spectrometry ◽  
Energy Source ◽  
Gas Chromatography ◽  
Benzene Ring ◽  
Metabolic Pathway ◽  
Mass Spectrometry Analysis ◽  
Gas Chromatography Mass Spectrometry ◽  
Spectrometry Analysis ◽  
Chromatography Mass Spectrometry ◽  
Dioxygenase Activity

ABSTRACT Burkholderia sp. strain AK-5 utilized 4-aminophenol as the sole carbon, nitrogen, and energy source. A pathway for the metabolism of 4-aminophenol in strain AK-5 was proposed based on the identification of three key metabolites by gas chromatography-mass spectrometry analysis. Strain AK-5 converted 4-aminophenol to 1,2,4-trihydroxybenzene via 1,4-benzenediol. 1,2,4-Trihydroxybenzene 1,2-dioxygenase cleaved the benzene ring of 1,2,4-trihydroxybenzene to form maleylacetic acid. The enzyme showed a high dioxygenase activity only for 1,2,4-trihydroxybenzene, with Km and V max values of 9.6 μM and 6.8 μmol min−1 mg of protein−1, respectively.

Isolation and Characterization of a Dibenzofuran-Degrading Yeast: Identification of Oxidation and Ring Cleavage Products

1998 ◽  
Vol 64 (6) ◽  
pp. 2215-2219 ◽  
Author(s):  
Elke Hammer ◽  
Dirk Krowas ◽  
Annett Schäfer ◽  
Michael Specht ◽  
Wittko Francke ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Performance ◽  
Analytical Data ◽  
Gas Chromatography Mass Spectrometry ◽  
Ring Cleavage ◽  
Resonance Spectroscopy ◽  
Yeast Identification ◽  
Isolation And Characterization ◽  
Incubation Experiments

ABSTRACT We characterized the ability of a yeast to cleave the aromatic structure of the dioxin-like compound dibenzofuran. The yeast strain was isolated from a dioxin-contaminated soil sample and identified asTrichosporon mucoides. During incubation of glucose-pregrown cells with dibenzofuran, six major metabolites were detected by high-performance liquid chromatography. The formation of four different monohydroxylated dibenzofurans was proven by comparison of analytical data (gas chromatography-mass spectrometry) with that for authentic standards. Further oxidation produced 2,3-dihydroxydibenzofuran and its ring cleavage product 2-(1-carboxy methylidene)-2,3-dihydrobenzo[b]furanylidene glycolic acid, which were characterized by mass spectrometry and 1H nuclear magnetic resonance spectroscopy. These two metabolites are derived from 2-hydroxydibenzofuran and 3-hydroxydibenzofuran, as shown by incubation experiments using these monohydroxylated dibenzofurans as substrates.

scholarly journals A Polyomic Approach To Elucidate the Fluoranthene-Degradative Pathway in Mycobacterium vanbaalenii PYR-1

2007 ◽  
Vol 189 (13) ◽  
pp. 4635-4647 ◽  
Author(s):  
Ohgew Kweon ◽  
Seong-Jae Kim ◽  
Richard C. Jones ◽  
James P. Freeman ◽  
Michael D. Adjei ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
High Performance ◽  
Culture Medium ◽  
Gas Chromatography Mass Spectrometry ◽  
Visible Absorption ◽  
Wide Range ◽  
Liquid Chromatography Tandem Mass ◽  
Polycyclic Aromatic ◽  
Uv Visible

ABSTRACT Mycobacterium vanbaalenii PYR-1 is capable of degrading a wide range of high-molecular-weight polycyclic aromatic hydrocarbons (PAHs), including fluoranthene. We used a combination of metabolomic, genomic, and proteomic technologies to investigate fluoranthene degradation in this strain. Thirty-seven fluoranthene metabolites including potential isomers were isolated from the culture medium and analyzed by high-performance liquid chromatography, gas chromatography-mass spectrometry, and UV-visible absorption. Total proteins were separated by one-dimensional gel and analyzed by liquid chromatography-tandem mass spectrometry in conjunction with the M. vanbaalenii PYR-1 genome sequence (http://jgi.doe.gov ), which resulted in the identification of 1,122 proteins. Among them, 53 enzymes were determined to be likely involved in fluoranthene degradation. We integrated the metabolic information with the genomic and proteomic results and proposed pathways for the degradation of fluoranthene. According to our hypothesis, the oxidation of fluoranthene is initiated by dioxygenation at the C-1,2, C-2,3, and C-7,8 positions. The C-1,2 and C-2,3 dioxygenation routes degrade fluoranthene via fluorene-type metabolites, whereas the C-7,8 routes oxidize fluoranthene via acenaphthylene-type metabolites. The major site of dioxygenation is the C-2,3 dioxygenation route, which consists of 18 enzymatic steps via 9-fluorenone-1-carboxylic acid and phthalate with the initial ring-hydroxylating oxygenase, NidA3B3, oxidizing fluoranthene to fluoranthene cis-2,3-dihydrodiol. Nonspecific monooxygenation of fluoranthene with subsequent O methylation of dihydroxyfluoranthene also occurs as a detoxification reaction.

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