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1.
《Journal of Fermentation and Bioengineering》1989,67(4):291-296
We have developed a simple method for the detection of phthalate 4,5-dioxygenase and 4,5-dihydro-4,5-dihydroxyphthalate dehydrogenase activities in the initial step of phthalate degradation in bacteria. It was found that cells of a Pseudomonas putida strain adapted for phthalate could convert quinolinic acid to a hydroxylated product having λmax at 315 nm. The occurrence of this compound was visualized by reaction with diazotized p-nitroaniline with which a red compound having λmax at 512 nm was produced. In practice, if cells in colonies developed on an agar plate containing mineral salt medium supplemented with 0.4% of disodium phthalate and 0.1% of quinolinic acid are active with respect to the 4,5-dihydroxyphthalate pathway, then the colonies would be colored red immediately upon spraying with the diazotized p-nitroaniline reagent. The method was used to identify the phthalate degradative pathway for 27 phthalate-utilizing strains of the genera Pseudomonas (18 strains), Agrobacterium (3 strains), Alcaligenes (5 strains), and Micrococcus (1 strain). It was found that 24 of the 26 Gram-negative bacteria have the 4,5-dihydroxyphthalate pathway and that the remaining two strains of Pseudomonas sp. may metabolize via an unidentified pathway other than the dihydroxyphthalate pathways, and the Gram-positive strain of Micrococcus sp. metabolizes phthalate via the 3,4-dihydroxyphthalate pathway. 相似文献
2.
Utilization of phthalate esters by micrococci 总被引:5,自引:0,他引:5
Several strains of Micrococcus have been isolated by enrichment with one of several phthalate esters as sole carbon source. They have been separated into four groups by their esterase content and nutritional characteristics. The catabolic potential for phthalate utilization found in these strains provides further support for designation of the four groups. Pathways for phthalate utilization by 4,5-dihydroxyphthalate and/or 3,4-dihydroxyphthalate and protocatechuate and/or 2,3-dihydroxybenzoate are outlined, which suggests that micrococci possess substantial potential for the catabolism of aromatic compounds. 相似文献
3.
Pseudomonas aeruginosa PP4, Pseudomonas sp. PPD and Acinetobacter lwoffii ISP4 capable of utilizing phthalate isomers were isolated from the soil using enrichment culture technique. The strain ISP4 metabolizes isophthalate, while PPD and PP4 utilizes all three phthalate isomers (ortho-, iso- and tere-) as the sole carbon source. ISP4 utilizes isophthalate (0.1%) more rapidly (doubling time, 0.9 h) compared to PPD (4.64 h), PP4 (7.91 h) and other reported strains so far. The metabolic pathways in these isolates were initiated by dihydroxylation of phthalate isomers. Phthalate is hydroxylated to 3,4-dihydro-3,4-dihydroxyphthalate and 4,5-dihydro-4,5-dihydroxyphthalate in strains PP4 and PPD, respectively; while terephthalate is hydroxylated to 2-hydro-1,2-dihydroxyterephthalate. All three strains hydroxylate isophthalate to 4-hydro-3,4-dihydroxyisophthalate. The generated dihydroxyphthalates were subsequently metabolized to 3,4-dihydroxybenzoate (3,4-DHB) which was further metabolized by ortho ring-cleavage pathway. PP4 and PPD cells grown on phthalate, isophthalate or terephthalate showed respiration on respective phthalate isomer and the activity of corresponding ring-hydroxylating dioxygenase, suggesting the carbon source specific induction of three different ring-hydroxylating dioxygenases. We report, for the first time, the activity of isophthalate dioxygenase and its reductase component in the cell-free extracts. The enzyme showed maximum activity with reduced nicotinamide adenine dinucleotide (NADH) in the pH range 8–8.5. Cells grown on glucose failed to respire on phthalate isomers and 3,4-DHB and showed significantly low activities of the enzymes suggesting that the enzymes are inducible. 相似文献
4.
Phthalate metabolism in Pseudomonas testosteroni: accumulation of 4,5-dihydroxyphthalate by a mutant strain. 总被引:6,自引:4,他引:2
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A mutant strain of Pseudomonas testosteroni blocked in phthalate catabolism converted phthalate into 4,5-dihydroxyphthalate. The latter compound was isolated, and its physical properties were determined. A stoichiometric conversion of the compound to protocatechuate was demonstrated spectrophotometrically with crude extracts of a protocatechuate 4,5-dioxygenase-deficient mutant. Therefore, phthalate is metabolized through 4,5-dihydroxyphthalate and protocatechuate, which is further degraded by protocatechuate 4,5-dioxygenase in P. testosteroni. By using several mutants blocked in phthalate catabolism, 4,5-dihydroxyphthalate decarboxylase was shown to be induced by phthalate. A simple spectrophotometric assay for the enzyme is also reported. 相似文献
5.
Toshio Omori Minoru Matsubara Susumu Masuda Tohru Kodama 《Applied microbiology and biotechnology》1991,35(4):431-435
Summary
Pseudomonas testosteroni M4-1, capable of using phthalate as the sole carbon and energy source, was isolated. Tn5 mutagenesis using pSUP2021 yielded mutant strains of M4-1 that are defective in phthalate metabolism and produce a dihydrodiol compound. The dihydrodiol compound produced by mutant strain M4-122 was isolated and identified as 4,5-dihydro-4,5-dihydroxyphthalate (DDP) by elementary analysis, mass analysis and nuclear magnetic resonance. Various conditions to increase the yield of DDP from phthalate were examined for mutant strain M4-122. With resting cells 6 g DDP/1 were produced. The additional of ethanol to the resting-cell reaction mixture enhanced DDP production and 10 g DDP/1 was produced from 8.3 g/1 of phthalate.
Offprint requests to: T. Omori 相似文献
6.
A. Suemori K. Nakajima R. Kurane Y. Nakamura 《Applied microbiology and biotechnology》1995,43(3):470-472
Gram-positive Rhodococcus erythropolis strain S1 formed enzymes for the degradation of phthalate when grown in a phthalate-containing minimal medium. The membrane
fraction prepared from phthalate-grown cells by ultrasonication converted phthalate to protocatechuate as the final product.
Using two membrane-bound enzymes, phthalate 3,4-dioxygenase (PO) and 3,4-dihydro-3,4-dihydroxyphthalate 3,4-dehydrogenase
(PH), prepared by solubilization of the membrane fraction, 3,4-dihydroxyphthalate was selectively obtained from phthalata.
Fe2+ and Mn2+ stimulated the formation of 3,4-dihydroxyphthalate by the membrane-bound PO and PH system.
Received: 27 April 1994/Received last revision: 19 August 1994/Accepted: 12 September 1994 相似文献
7.
The decarboxylation of phthalic acids was studied with Bacillus sp. strain FO, a marine mixed culture ON-7, and Pseudomonas testosteroni. The mixed culture ON-7, when grown anaerobically on phthalate but incubated aerobically with chloramphenicol, quantitatively converted phthalic acid to benzoic acid. Substituted phthalic acids were also decarboxylated: 4,5-dihydroxyphthalic acid to protocatechuic acid; 4-hydroxyphthalic and 4-chlorophthalic acids to 3-hydroxybenzoic and 3-chlorobenzoic acids, respectively; and 3-fluorophthalic acid to 2-and 3-fluorobenzoic acids. Bacillus sp. strain FO gave similar results except that 4,5-dihydroxyphthalic acid was not metabolized, and both 3- and 4-hydroxybenzoic acids were produced from 4-hydroxyphthalic acid. P. testosteroni decarboxylated 4-hydroxyphthalate (to 3-hydroxybenzoate) and 4,5-dihydroxyphthalate but not phthalic acid and halogenated phthalates. Thus, P. testosteroni and the mixed culture ON-7 possessed 4,5-dihydroxyphthalic acid decarboxylase, previously described in P. testosteroni, that metabolized 4,5-dihydroxyphthalic acid and specifically decarboxylated 4-hydroxyphthalic acid to 3-hydroxybenzoic acid. The mixed culture ON-7 and Bacillus sp. strain FO also possessed a novel decarboxylase that metabolized phthalic acid and halogenated phthalates, but not 4,5-dihydroxyphthalate, and randomly decarboxylated 4-hydroxyphthalic acid. The decarboxylation of phthalic acid is suggested to involve an initial reduction to 1,2-dihydrophthalic acid followed by oxidative decarboxylation to benzoic acid. 相似文献
8.
Metabolism of dibutylphthalate and phthalate by Micrococcus sp. strain 12B. 总被引:14,自引:10,他引:4
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Micrococcus sp. strain 12B was isolated by enriching for growth with dibutylphthalate as the sole carbon and energy source. A pathway for the metabolism of dibutylphthalate and phthalate by micrococcus sp. strain 12B is proposed: dibutylphthalate leads to monobutylphthalate leads to phthalate leads to 3,4-dihydro-3,4-dihydroxyphthalate leads to 3,4-dihydroxyphthalate leads to protocatechuate (3,4-dihdroxybenzoate). Protocatechuate is metabolized both by the meta-cleavage pathway through 4-carboxy-2-hydroxymuconic semialdehyde and 4-carboxy-2-hydroxymuconate to pyruvate and oxaloacetate and by the ortho-cleavage pathway to beta-ketoadipate. Dibutylphthalate- and phthalate-grown cells readily oxidized dibutylphthalate, phthalate, 3,4-dihydroxyphthalate, and protocatechuate. Extracts of cells grown with dibutylphthalate or phthalate contained the 3,4-dihydroxyphthalate decarboxylase and the enzymes of the protocatechuater 4,5-meta-cleavage pathway. Extracts of dibutylphthalate-grown cells also contained the protocatechuate ortho-cleavage pathway enzymes. The dibutylphthalate-hydrolyzing esterase and 3,4-dihydroxyphthalate decarboxylase were constitutively synthesized; phthalate-3,4-dioxygenase (and possibly the "dihydrodiol" dehydrogenase) was inducible by phthalate or a metabolite occurring before protocatechuate in the pathway; two protocatechuate oxygenases and subsequent enzymes were inducible by protocatechuate or a subsequent metabolic product. During growth at 37 degrees C, strain 12B gave clones at high frequency that had lost the ability to grow with phthalate esters. One of these nonrevertible mutants, strain 12B-Cl, lacked all of the enzymes required for the metabolism of dibutylphthalate through the protocatechuate meta-cleavage pathway. Enzymes for the metabolism of protocatechuate by the ortho-cleavage pathway were present in this strain grown with p-hydroxybenzoate or protocatechuate. 相似文献
9.
Tina L. T. Gresham Peter P. Sheridan Mary E. Watwood Yoshiko Fujita Frederick S. Colwell 《Geomicrobiology journal》2013,30(3-4):353-364
Polymerase chain reaction primers based on the ureC gene are described for use in detecting diverse groundwater urea-hydrolyzing bacteria. Six degenerate primers were designed and evaluated for their ability to detect the gene encoding the large catalytic subunit of urease, ureC. Five combinations of these primers were tested pair-wise and displayed an overlapping detection range for bacterial isolates. Pair L2F/L2R exhibited the greatest detection range for described bacterial species and for bacterial isolates from groundwater samples belonging to the bacterial divisions Firmicutes, Actinobacteria, and the α , β , and γ subdivisions of Proteobacteria. Primers L2F/L2R exhibited a greater detection range than previously described ureC-specific primers, and amplified novel ureC sequences from groundwater isolates in the genera Hydrogenophaga, Acidovorax, Janthinobacterium, and Arthrobacter. A comparative phylogenetic analysis of ureC and 16S rRNA genes was performed to determine the utility of groundwater ureC sequence information as a phylogenetic marker for ureolytic species. Our results were consistent with previous analyses of urease genes which demonstrated that the ureC gene has undergone lateral transfer and is not a robust phylogenetic marker. However, the ureC-specific primers, L2F/L2R, demonstrate a broad detection range for ureolytic species, and can serve to enhance functional diversity analyses of ureolytic bacteria. 相似文献
10.
Micrococcus strain 12B, grown with phthalate, transformed the phthalate analog, phthalaldehydate (2-formylbenzoate), to 3,4-dihydroxyphthalaldehydate which was isolated and identified as its lactol. An 18O2 incorporation experiment indicated that a dioxygenase mechanism was involved. It is proposed by analogy, that phthalate is metabolized through cis-3,4-dihydro-3,4-dihydroxyphthalate and 3,4-dihydroxyphthalate by this bacterium. 相似文献
11.
Shinohara H Yoshida S Enya J Watanabe Y Tsukiboshi T Negishi H Tsushima S 《Folia microbiologica》2011,56(6):505-517
Culturable bacterial communities on rice plants were investigated from 2001 to 2003. In total, 1,394 bacterial isolates were
obtained from the uppermost leaf sheaths at 1 month before heading time and from leaf sheaths and panicles at heading time.
The average culturable bacterial population on the leaf sheaths was larger at heading time than at 1 month previously. Furthermore,
the population was significantly larger on panicles than on leaf sheaths, suggesting that the bacterial population is influenced
by the organs of rice plants. Larger proportions of bacteria were obtained from the macerates of leaf sheaths after washing
with phosphate buffer, and most culturable bacteria were verified to inhabit the inside or inner surface, rather than the
outer surface, of the tissues. Verification of the bacterial composition based on 16S rRNA gene sequences revealed that genera
of Sphingomonas, Microbacterium, Methylobacterium, and Acidovorax tended to be dominant colonizers on leaf sheaths, whereas Pseudomonas and Pantoea were isolated mainly from the panicles, indicating that leaf sheaths and panicles harbor distinct communities. Furthermore,
the richness of bacterial genera was less on both leaf sheaths and panicles at heading time compared with that observed 1 month
before heading time. Phylogenetic analyses using bacterial isolates belonging to the four dominant genera inhabiting leaf
sheaths at heading time revealed that particular bacterial groups in each genus colonized the leaf sheaths. 相似文献
12.
Joshi AA Kanekar PP Kelkar AS Shouche YS Vani AA Borgave SB Sarnaik SS 《Microbial ecology》2008,55(2):163-172
Aerobic, alkaliphilic bacteria were isolated and characterized from water and sediment samples collected in the winter season,
January 2002 from alkaline Lonar lake, India, having pH 10.5. The total number of microorganisms in the sediment and water
samples was found to be 102–106 cfu g−1 and 102–104 cfu ml−1, respectively. One hundred and ninety-six strains were isolated using different enrichment media. To study the bacterial
diversity of Lonar lake and to select the bacterial strains for further characterization, screening was done on the basis
of pH and salt tolerance of the isolates. Sixty-four isolates were subjected to phenotypic, biochemical characterization and
16S rRNA sequencing. Out of 64, 31 bacterial isolates were selected on the basis of their enzyme profile and further subjected
to phylogenetic analysis. Phylogenetic analysis indicated that most of the Lonar lake isolates were related to the phylum
Firmicutes, containing Low G+C, Gram-positive bacteria, with different genera: Bacillus, Paenibacillus, Alkalibacillus, Exiguobacterium, Planococcus, Enterococcus and Vagococcus. Seven strains constituted a Gram-negative bacterial group, with different genera: Halomonas, Stenotrophomonas and Providencia affiliated to γ-Proteobacteria, Alcaligenes to β-Proteobacteria and Paracoccus to α-Proteobacteria. Only five isolates were High G+C, Gram-positive bacteria associated with phylum Actinobacteria, with
various genera: Cellulosimicrobium, Dietzia, Arthrobacter and Micrococcus. Despite the alkaline pH of the Lonar lake, most of the strains were alkalitolerant and only two strains were obligate alkaliphilic.
Most of the isolates produced biotechnologically important enzymes at alkaline pH, while only two isolates (ARI 351 and ARI
341) showed the presence of polyhydroxyalkcanoate (PHA) and exopolysaccharide (EPS), respectively. 相似文献
13.
Ryuichiro Kurane Tomoo Suzuki Yoshimasa Takahara Kazuo Komagata 《Bioscience, biotechnology, and biochemistry》2013,77(6):1031-1038
Seventeen bacterial capable of utilizing phthalate esters isolated from natural sources were identified. Based on morphological and biochemical characteristics, type of cell division, GC content in DNA, principal amino acid in the cell wall and cellular fatty acid composition, 10 isolates were identified as Nocardia erythropolis, one isolate as Pseudomonas acidovorans, another as Pseudomonas cepacia and four as members of the genus of Corynebacterium. 相似文献
14.
《Bioscience, biotechnology, and biochemistry》2013,77(9):1482-1483
Phthalate oxygenase was induced in Rhodococcus erythropolis S-1, a Gram-positive bacterium, when this bacterium was cultured in a medium containing phthalate as a sole carbon source. The enzyme was purified 118-fold with 4.7% activity yield. The purified enzyme appeared homogenous on native PAGE. This enzyme is a large protein (213 kDa), a tetramer of identical 56kDa monomers, and a flavoprotein containing FAD with NADH-dependent dioxygenase activity. The enzyme is specific for phthalate and other closely related aromatic compounds. Optimum pH and temperature were 6.5 and 40°C. The Km for phthalate and NADH were 0.040 mM and 0.069 mM. The enzyme catalyzes dihydroxylation of phthalate to form 3,4-dihydro-3,4-dihydroxyphthalate with consumption of NADH and oxygen. 相似文献
15.
Tri Joko Ahmad Subandi Nanda Kusumandari Arif Wibowo Achmadi Priyatmojo 《Archives Of Phytopathology And Plant Protection》2013,46(10):1239-1250
Soft rot by bacterial pathogens is one of the most widespread and destructive diseases on various plants including orchids throughout the world. The pathogenicity of the pathogens is reported to be mainly determined by massive production of plant cell wall-degrading enzymes (PCDE). In the previous work, we have isolated 20 isolates of bacterial soft rot from orchids collected in Yogyakarta Special Region and West Java province, Indonesia. In this study, we further confirmed them as pathogens by hypersensitive reaction assay on tobacco leaves followed by pathogenicity test on Phalaenopsis sp. The production of four major PCDE by qualitative plate assays including pectate lyase, polygalacturonase, cellulase and protease was also evaluated. Even though all the isolates were able to initiate soft rot symptom, our results showed two distinct groups which clustered as producing and non-producing PCDE. The 16S rDNA analysis revealed that the isolates belonged to the genera Pectobacterium, Klebsiella, Serratia, Enterobacter, Citrobacter, Providencia and Pseudomonas. 相似文献
16.
The bacteriumPseudomonas sp. strain RW31 isolated from the river Elbe utilized the ammonium salt of 4-sulfophthalate (4SPA) as sole source of carbon, sulfur, nitrogen, and energy and grew also with phthalate (PA) and several other aromatic compounds as sole carbon and energy source. The xenobiotic sulfo group of 4SPA was eliminated as sulfite, which transiently accumulated in the culture supernatant up to about 10 µM and was slowly oxidized to the stoichiometrical amount of sulfate. Biodegradation routes of 4SPA as well as of PA converged into the protocatechuate pathway and from found activities for the decarboxylation of 4,5-dihydroxyphthalate we deduce this compound the first rearomaticized intermediate after initial dioxygenation. Protocatechuate then underwentmeta-cleavage mediated by a protocatechuate 4,5-dioxygenase activity which was competitively inhibited by the structurally related compound 3,4,5-trihydroxybenzoate; protocatechuate accumulated in the medium up to an about 2 mM concentration. Indications for the presence of selective transport systems are presented. 相似文献
17.
Renxing Liang Xueling Wu Qinyun Dai Decai Jin Yangyang Wang 《Antonie van Leeuwenhoek》2010,97(1):79-89
Thirty-two strains of phthalic acid ester (PAEs)-degrading bacteria were isolated from thirteen geographically diverse sites
by enrichment using mixtures of PAEs as the sole source of carbon and energy. Sequence analyses of the 16S rRNA gene indicated
that these isolates were from six genera (Arthrobacter, Gordonia, Rhodococcus, Acinetobacter, Pseudomonas, and Delftia). To evaluate the genetic diversity among them, the molecular typing method rep-PCR with primers based on enterobacterial
repetitive intergenic consensus, repetitive extragenic palindromes, and BOXAIR sequences was performed. Strain-specific and
unique genotypic fingerprints were distinguished for most of these isolates. In addition, utilization of various PAEs and
the central intermediate phthalic acid by representative isolates suggested inter-isolate differences in the substrate utilization
and degradation pathways. Furthermore, HPLC analysis showed that the rate of dimethyl phthalate degradation varied from 48.32
to 100% between strains. These results suggest a high level of genetic diversity among PAEs-degrading bacteria in the natural
environment and their great potential to clean up phthalates-contaminated environments. 相似文献
18.
For a long time it was thought that Bursaphelenchus xylophilus was the only agent of the pine wilt disease. Recently, it was discovered that there are bacteria associated with the nematodes that contribute to the pathogenesis of this disease, mainly through the release of toxins that promote the death of the pines. Among the species most commonly found, are bacteria belonging to the Bacillus, Pantoea, Pseudomonas and Xanthomonas genera.The main objective of this work was to study the effect of inoculation of maritime pine (Pinus pinaster) with four different nematode isolates, in the bacterial population of nematodes and trees, at different stages of disease progression. The monitoring of progression of disease symptoms was also recorded. Also, the identification of bacteria isolated from the xylem of trees and the surface of nematodes was performed by classical identification methods, by the API20E identification system and by sequencing of bacterial DNA.The results showed that for the symptoms progression, the most striking difference was observed for the pines inoculated with the avirulent isolate, C14-5, which led to a slower and less severe aggravation of symptoms than in pines inoculated with the virulent isolates. In general, it was found that bacterial population, inside the tree, increased with disease progression. A superior bacterial quantity was isolated from pines inoculated with the nematode isolates HF and 20, and, comparatively, few bacteria were isolated from pines inoculated with the avirulent isolate. The identification system API20E was insufficient in the identification of bacterial species; Enterobacter cloacae species was identified in 79% of the isolated bacterial colonies and seven of these colonies could not be identified by this method. Molecular identification methods, through bacterial DNA sequencing, allowed a more reliable identification: eleven different bacterial species within the Bacillus, Citrobacter, Enterobacter, Escherichia, Klebsiella, Paenibacillus, Pantoea and Terribacillus genera were identified. General bacterial diversity increased with the progression of the disease. Bacillus spp. were predominant at the earlier stage of disease progression and Klebsiella oxytoca at the later stages. Furthermore, bacterial species isolated from the surface of nematodes were similar to those isolated from the xylem of pines.In the present work new bacterial species were identified which have never been reported before in this type of study and may be associated with their geographical origin (Portugal). P. pinaster, the pine species used in this study, was different from those commonly grown in Japan and China. Furthermore, it was the first time that bacteria were isolated and identified from an avirulent pine wood nematode isolate. 相似文献
19.
Yoshiaki Maejima Koya Kushimoto Yusuke Muraguchi Kohei Fukuda Takamasa Miura Atsushi Yamazoe 《Bioscience, biotechnology, and biochemistry》2018,82(7):1260-1263
141 filterable bacteria that passed through a 0.22 μm pore size filter were isolated from Lake Sanaru in Hamamatsu, Japan. These belonged to Proteobacteria, Bacteroidetes, Firmicutes, or Actinobacteria among which the first two phyla comprised the majority of the isolates. 48 isolates (12 taxa) are candidates assignable to new bacterial species or genera of Proteobacteria or Bacteroidetes. 相似文献
20.
Using protocols designed for the isolation of Shigella from environmental freshwater samples from different regions of Bangladesh, 11 bacterial strains giving rise to Shigella-like colonies on selective agar plates and showing serological cross-reaction with Shigella-specific antisera were isolated. Phylogenetic analyses revealed that three of the isolates were most closely related to Escherichia coli, four to Enterobacter sp., two to Stenotrophomonas, and two isolates belonged to the Gram-positive genus Aerococcus. The isolates cross-reacted with six different serotypes of Shigella and were, in each case, highly type-specific. Two of the isolates belonging to the Enterobacter and Escherichia genera gave extremely strong cross-reactivity with Shigella dysenteriae and Shigella boydii antisera, respectively. The Aerococcus isolates gave relatively weak but significant cross-reactions with S. dysenteriae. Western blot analysis revealed that a number of antigens from the isolates cross-react with Shigella spp. The results indicate that important Shigella spp. surface antigens are shared by a number of environmental bacteria, which have implications for the use of serological
methods in attempts for the detection and recovery of Shigella from aquatic environments. 相似文献