scholarly journals Efficient Degradation of 2,4,6-Trichlorophenol Requires a Set of Catabolic Genes Related to tcp Genes from Ralstonia eutropha JMP134(pJP4)

2003 ◽  
Vol 69 (12) ◽  
pp. 7108-7115 ◽  
Author(s):  
V. Matus ◽  
M. A. Sánchez ◽  
M. Martínez ◽  
B. González

ABSTRACT 2,4,6-Trichlorophenol (2,4,6-TCP) is a hazardous pollutant. Several aerobic bacteria are known to degrade this compound. One of these, Ralstonia eutropha JMP134(pJP4), a well-known, versatile chloroaromatic compound degrader, is able to grow in 2,4,6-TCP by converting it to 2,6-dichlorohydroquinone, 6-chlorohydroxyquinol, 2-chloromaleylacetate, maleylacetate, and β-ketoadipate. Three enzyme activities encoded by tcp genes, 2,4,6-TCP monooxygenase (tcpA), 6-chlorohydroxyquinol 1,2-dioxygenase (tcpC), and maleylacetate reductase (tcpD), are involved in this catabolic pathway. Here we provide evidence that all these tcp genes are clustered in the R. eutropha JMP134(pJP4) chromosome, forming the putative catabolic operon tcpRXABCYD. We studied the presence of tcp-like gene sequences in several other 2,4,6-TCP-degrading bacterial strains and found two types of strains. One type includes strains belonging to the Ralstonia genus and possessing a set of tcp-like genes, which efficiently degrade 2,4,6-TCP and therefore grow in liquid cultures containing this chlorophenol as a sole carbon source. The other type includes strains belonging to the genera Pseudomonas, Sphingomonas, or Sphingopixis, which do not have tcp-like gene sequences and degrade this pollutant less efficiently and which therefore grow only as small colonies on plates with 2,4,6-TCP. Other than strain JMP134, none of the bacterial strains whose genomes have been sequenced possesses a full set of tcp-like gene sequences.

2000 ◽  
Vol 66 (4) ◽  
pp. 1602-1608 ◽  
Author(s):  
D. Pérez-Pantoja ◽  
L. Guzmán ◽  
M. Manzano ◽  
D. H. Pieper ◽  
B. González

ABSTRACT The enzymes chlorocatechol-1,2-dioxygenase, chloromuconate cycloisomerase, dienelactone hydrolase, and maleylacetate reductase allow Ralstonia eutropha JMP134(pJP4) to degrade chlorocatechols formed during growth in 2,4-dichlorophenoxyacetate or 3-chlorobenzoate (3-CB). There are two gene modules located in plasmid pJP4,tfdCIDIEIFI (module I) andtfdDIICIIEIIFII (module II), putatively encoding these enzymes. To assess the role of both tfd modules in the degradation of chloroaromatics, each module was cloned into the medium-copy-number plasmid vector pBBR1MCS-2 under the control of the tfdR regulatory gene. These constructs were introduced into R. eutropha JMP222 (a JMP134 derivative lacking pJP4) and Pseudomonas putidaKT2442, two strains able to transform 3-CB into chlorocatechols. Specific activities in cell extracts of chlorocatechol-1,2-dioxygenase (tfdC), chloromuconate cycloisomerase (tfdD), and dienelactone hydrolase (tfdE) were 2 to 50 times higher for microorganisms containing module I compared to those containing module II. In contrast, a significantly (50-fold) higher activity of maleylacetate reductase (tfdF) was observed in cell extracts of microorganisms containing module II compared to module I. The R. eutropha JMP222 derivative containingtfdR-tfdCIDIEIFI grew four times faster in liquid cultures with 3-CB as a sole carbon and energy source than in cultures containingtfdR-tfdDIICIIEIIFII . In the case of P. putida KT2442, only the derivative containing module I was able to grow in liquid cultures of 3-CB. These results indicate that efficient degradation of 3-CB by R. eutropha JMP134(pJP4) requires the two tfd modules such that TfdCDE is likely supplied primarily by module I, while TfdF is likely supplied by module II.


2001 ◽  
Vol 67 (1) ◽  
pp. 42-50 ◽  
Author(s):  
Dirk Springael ◽  
Annemie Ryngaert ◽  
Christophe Merlin ◽  
Ariane Toussaint ◽  
Max Mergeay

ABSTRACT Tn4371, a 55-kb transposable element involved in the degradation and biphenyl or 4-chlorobiphenyl identified inRalstonia eutropha A5, displays a modular structure including a phage-like integrase gene (int), aPseudomonas-like (chloro)biphenyl catabolic gene cluster (bph), and RP4- and Ti-plasmid-like transfer genes (trb) (C. Merlin, D. Springael, and A. Toussaint, Plasmid 41:40–54, 1999). Southern blot hybridization was used to examine the presence of different regions of Tn4371 in a collection of (chloro)biphenyl-degrading bacteria originating from different habitats and belonging to different bacterial genera. Tn4371-related sequences were never detected on endogenous plasmids. Although the gene probes containing only bph sequences hybridized to genomic DNA from most strains tested, a limited selection of strains, all β-proteobacteria, displayed hybridization patterns similar to the Tn4371 bph cluster. Homology between Tn4371 and DNA of two of those strains, originating from the same area as strain A5, extended outside the catabolic genes and covered the putative transfer region of Tn4371. On the other hand, none of the (chloro)biphenyl degraders hybridized with the outer left part of Tn4371 containing the int gene. Thebph catabolic determinant of the two strains displaying homology to the Tn4371 transfer genes and a third strain isolated from the A5 area could be mobilized to a R. eutropha recipient, after insertion into an endogenous or introduced IncP1 plasmid. The mobilized DNA of those strains included all Tn4371 homologous sequences previously identified in their genome. Our observations show that the bph genes present on Tn4371 are highly conserved between different (chloro)biphenyl-degrading hosts, isolated globally but belonging mainly to the β-proteobacteria. On the other hand, Tn4371-related mobile elements carrying bphgenes are apparently only found in isolates from the environment that provided the Tn4371-bearing isolate A5.


1997 ◽  
Vol 60 (6) ◽  
pp. 639-643 ◽  
Author(s):  
FUENG-LIN KUO ◽  
JOHN B. CAREY ◽  
STEVEN C. RICKE

The effects were investigated of 254-nm UV radiation on populations of Salmonella typhimurium, aerobes, and molds on the shells of eggs. In the first experiment, the CFU of attached S. typhimurium cells on unwashed clean shell eggs were determined after 0, 1, 3, 5, and 7 min of UV treatment (620 μW/cm2) on both ends of the egg. All UV treatments significantly reduced S. typhimurium CFU (P < .01). UVtreatment (620 μW/cm2) in 1-min alternating light and dark cycles for 5 min (three light and two dark) was compared to 0, 3, and 5 min of UV treatment. No significant differences in microbial populations were observed among light and dark cycles and the other UV treatments. In a subsequent experiment, the same UV treatments were utilized to evaluate photoreactivation. After UV exposure, eggs were exposed to 1 h of fluorescent light or I h of darkness or cultured immediately. S. typhimurium CFU were significantly (P < .01) reduced by the UV treatments. However, no significant differences between microbial populations exposed to UV treatment and UV radiation plus photoreactivation were detected. For studies of aerobic bacteria and molds, different UV treatment times (0, 15, and 30 min) at the intensity of 620 μW/cm2 and different intensities (620, 1350, and 1720 μW/cm2) for 15 min were evaluated. Mold CFU per egg were either 0 or 1 for all UV treatments and a 99% reduction of CFU of aerobic bacteria per egg were observed for all UV treatments. It appears from these studies that UV light can significantly reduce populations of S. typhimurium, aerobes, and molds on shell eggs.


Plant Disease ◽  
2015 ◽  
Vol 99 (2) ◽  
pp. 281-281 ◽  
Author(s):  
V. Stojšin ◽  
J. Balaž ◽  
D. Budakov ◽  
Slaviša Stanković ◽  
I. Nikolić ◽  
...  

A severe bacterial leaf spot was observed during June and July 2013 on commercial cultivars of sugar beet (Beta vulgaris var. saccharifera) in the Vojvodina Province of Serbia. Serbia is a major sugar beet production area in southeastern Europe, with 62,895 ha and 3 million tons of sugar beet yield in 2013. A foliar leaf spot observed in 25 commercial sugar beet fields surveyed ranged from 0.1 to 40% severity. Symptoms were characterized as circular or irregular, 5- to 20-mm diameter, white to light brown necrotic spots, each with a dark margin. Diseased leaves were rinsed in sterilized, distilled water (SDW) and dried at room temperature, and leaf sections taken from the margin of necrotic tissue were macerated in SDW. Isolations from 48 symptomatic leaves onto nutrient agar with 5% (w/v) sucrose (NAS) produced bacterial colonies that were whitish, circular, dome-shaped, and Levan-positive. Representative isolates (n = 105) were Gram negative; aerobic; positive for catalase, fluorescence on King's medium B, and tobacco hypersensitivity; and negative for oxidase, potato rot, and arginine dehydrolase. These reactions corresponded to LOPAT group Ia, which includes Pseudomonas syringae pathovars (2). Repetitive extragenic palindromic sequence (rep)-PCR was used for genetic fingerprinting the isolates using the REP, ERIC, and BOX primers. Twenty-five different profiles were obtained among the strains. From each profile group, one representative strain was sequenced for the gyrB gene (1). Four heterogenic groups were observed, and representative gyrB gene sequences of each group were deposited in the NCBI GenBank (Accession Nos. KJ950024 to KJ950027). The sequences were compared with those of pathotype strain P. syringae pv. aptata CFBP 1617 deposited in the PAMDB database; one strain was 100% homologous, and the other three were 99% homologous. To fulfill identification of the Serbian sugar beet isolates, gltA and rpoD partial gene sequences were determined (1), and the sequences were deposited as Accession Nos. KM386838 to KM386841 for gltA and KM386830 to KM38683033 for rpoD. The sequences were 100% homologous with those of pathotype strain CFBP 1617. Pathogenicity of each of four representative bacterial strains was tested on 3-week-old plants of the sugar beet cultivars Marinela, Serenada, and Jasmina (KWS, Belgrade, Serbia) and Lara (NS Seme, Novi Sad, Serbia) by atomizing a bacterial suspension of ~106 CFU/ml of the appropriate isolate onto the abaxial leaf surface of three plants per cultivar until water-soaking of the leaf surface was observed. Three plants of each cultivar atomized similarly with P. syringae pv. aptata CFBP 2473 and SDW served as positive and negative control treatments, respectively. Inoculated plants were kept in a clear plastic box at 80 to 100% RH and 17 ± 1°C and examined for symptom development over 3 weeks. For all test isolates and the control strain, inoculated leaves first developed water-soaked lesions 7 days after inoculation (DAI). By 10 to 14 DAI, lesions were necrotic and infection had spread to the petioles. By 21 DAI, wilting was observed on more than 50% of inoculated plants. Negative control plants were symptomless. Bacteria re-isolated onto NAS from inoculated leaves had the same colony morphology, LOPAT results, and gyrB partial gene sequences as described for the test strains. No bacteria were re-isolated from negative control plants. Based on these tests, the pathogen causing leaf spot on sugar beet in Serbia was identified as P. syringae pv. aptata. References: (1) P. Ferrente and M. Scortichini. Plant Pathol. 59:954, 2010. (2) R. A. Lelliott et al. J. Appl. Bacteriol. 29:470, 1966.


2001 ◽  
Vol 44 (4) ◽  
pp. 373-381 ◽  
Author(s):  
José Américo de Sousa ◽  
Ângela Teresa Silva-Souza

A bacteriological study was conducted on fish and water from Congonhas River, Sertaneja (22º58’ S; 50º58’ W), Paraná State, Brazil. From 44% of the analysed fish, bacteria belonging to Pseudomonas, Acinetobacter, Aeromonas, Enterobacteriaceae, Micrococcus, Bacillus and Lactobacillus were isolated. The group most frequently isolated from fish was Aeromonas. In the water, the bacterial groups detected were Pseudomonas, Acinetobacter, Aeromonas, Enterobacteriaceae, Bacillus and Flavobacterium, from which Flavobacterium and Acinetobacter were the most abundant. The numbers of Colony Forming Units per millilitre of water varied from 3.1x10² to 1.0 x 10³. Although a clear pattern was not detected in the susceptibilities/resistances of the isolated strains to nine antimicrobial substances, Gram negative aerobic bacteria were more resistant than the other strains. A simultaneous resistance to furazolidone, oxolinic acid and norfloxacin, particularly in the bacteria isolated from fish, as well as in the aerobic strains isolated from water was observed. The antimicrobial substances to which less resistances were found were oxytetracycline in the strains isolated from water, and trimethoprim-sulphamethoxazole, oxytetracycline and chloramphenicol in those isolated from fish.


2022 ◽  
Vol 10 (1) ◽  
pp. 184
Author(s):  
Izumi Orita ◽  
Gento Unno ◽  
Risa Kato ◽  
Toshiaki Fukui

Methylorubrum extorquens AM1 is the attractive platform for the production of value-added products from methanol. We previously demonstrated that M. extorquens equipped with PHA synthase with broad substrate specificity synthesized polyhydroxyalkanoates (PHAs) composed of (R)-3-hydroxybutyrate and small fraction of (R)-3-hydroxyvalerate (3HV) and (R)-3-hydroxyhexanoate (3HHx) units on methanol. This study further engineered M. extorquens for biosynthesis of PHAs with higher 3HV and 3HHx composition focusing on the EMC pathway involved in C1 assimilation. The introduction of ethylmalonyl-CoA decarboxylase, catalyzing a backward reaction in the EMC pathway, aiming to increase intracellular propionyl/butyryl-CoA precursors did not affect PHA composition. Reverse b-oxidation pathway and subsequent (R)-specific hydration of 2-enoyl-CoA were then enhanced by heterologous expression of four genes derived from Ralstonia eutropha for the conversion of propionyl/butyryl-CoAs to the corresponding (R)-3-hydroxyacyl-CoA monomers. The resulting strains produced PHAs with higher 3HV and 3HHx compositions, while the methylotrophic growth was severely impaired. This growth impairment was interestingly restored by the addition of La3+ without a negative impact on PHA biosynthesis, suggesting the activation of the EMC pathway by La3+. The engineered M. extorquens synthesized PHA terpolymer composed of 5.4 mol% 3HV and 0.9% of 3HHx with 41% content from methanol as a sole carbon source in the presence of La3+.


2021 ◽  
Vol 9 (10) ◽  
pp. 2039
Author(s):  
Yura Choi ◽  
Shambhunath Bose ◽  
Jaegu Seo ◽  
Joo-Hyun Shin ◽  
Dokyung Lee ◽  
...  

Akkermansia muciniphila (A. muciniphila) is a promising probiotic candidate owing to its health-promoting properties. A previous study reported that the pasteurized form of A. muciniphila strains isolated from human stool samples had a beneficial impact on high-fat diet-induced obese mice. On the other hand, the differences in the probiotic effects between live and pasteurized A. muciniphila on the metabolism and immune system of the host are still inconclusive. This study examines the differences between the live and pasteurized forms of A. muciniphila strains on the lipid and glucose metabolism and on regulating the inflammatory immune responses using a HFD-fed obese mouse model. The animals were administered the live and pasteurized forms of two A. muciniphila strains five times per week for the entire study period of 12 weeks. Both forms of the bacterial strains improved the HFD-induced obesity and metabolic dysregulation in the mice by preventing body-weight gains after one week. In addition, they cause a decrease in the weights of the major adipose tissues, adipogenesis/lipogenesis and serum TC levels, improvement in glucose homeostasis and suppression of inflammatory insults. Furthermore, these treatments restored the damaged gut architecture and integrity and improved the hepatic structure and function in HFD-induced animals. On the other hand, for both bacterial strains, the pasteurized form was more potent in improving glucose tolerance than the live form. Moreover, specific A. muciniphila preparations with either live or pasteurized bacteria decreased the number and population (%) of splenic Treg cells (CD4+ Foxp3+) significantly in the HFD-fed animals, further supporting the anti-inflammatory properties of these bacteria.


1961 ◽  
Vol 155 (959) ◽  
pp. 195-201 ◽  

The lag preceding growth of Bact. lactis aerogenes (Aerobacter aerogenes) after a first transfer to a medium containing D-arabinose as sole carbon source increases with the age and decreases with the size of the inoculum. During the long lag phase the β -galactosidase activity declines steeply. In contrast with this (and with a control ageing in a glucose medium) the D-ribulose isomerase activity is maintained, although no detectable consumption of D-arabinose occurs. If the long lag of unadapted cells in D-arabinose is divided into parts by intermediate passages in glucose or lactose media, the sum of the partial lags is nearly constant and equal to that observed when there is no interruption. But the periodic passages in the other media increase the rate at which growth eventually occurs in the D-arabinose. It is concluded that during the lag a decay of the enzymes in general occurs concomitantly with the development of the specific mechanisms concerned in the utilization of the new substrate. The balance of these processes (together with varying loss or retention of diffusible metabolites) is largely responsible for the observed variations in lag and mean generation time.


2016 ◽  
Vol 27 (1) ◽  
pp. 44-47 ◽  
Author(s):  
Mihaela Carmen Eremia ◽  
Irina Lupescu ◽  
Mariana Vladu ◽  
Maria Petrescu ◽  
Gabriela Savoiu ◽  
...  

Abstract Polyhydroxyalcanoates (PHAs) are specifically produced by a wide variety of bacteria, as an intracellular energy reserve in the form of homo- and copolymers of [R]-β-hydroxyalkanoic acids, depending on the C source used for microorganism growth, when the cells are grown under stressing conditions. In this paper we present microbiological accumulation of poly-3-hydroxyoctanoate (PHO) by using a consortium of bacterial strains, Pseudomonas putida and Bacillus subtilis, in a rate of 3:1, grown on a fermentation medium based on sodium octanoate as the sole carbon source. The experiments performed in the above mentioned conditions led to the following results: from 18.70 g sodium octanoate (7.72 g/L in the fermentation medium) used up during the bioprocess, 3.93-3.96 g/L dry bacterial biomass and 1.834 - 1.884 g/L PHA, containing 85.83 - 86.8% PHO, were obtained.


Author(s):  
Hiroyuki Sawada ◽  
Takashi Fujikawa ◽  
Shigeru Osada ◽  
Mamoru Satou

Five phytopathogenic bacterial strains, MAFF 301449T, MAFF 301450, MAFF 301451, MAFF 301452, and MAFF 301453, which were isolated from bud blight lesions of cyclamen (Cyclamen persicum Mill.) in Miyagi, Japan, were subjected to polyphasic taxonomic characterisation. The cells were Gram-reaction-negative, aerobic, non-spore-forming, motile with one to five polar flagella, and rod-shaped. Analysis of 16S rRNA gene sequences showed that they belong to the genus Pseudomonas , with Pseudomonas extremaustralis 14-3T (99.79 % sequence similarity), Pseudomonas trivialis DSM 14937T (99.79 %), Pseudomonas poae DSM 14936T (99.72 %), and Pseudomonas antarctica CMS 35T (99.72 %) as their relatives. The genomic DNA G+C content was 60.3 mol% and the major fatty acids (>5 % of the total fatty acids) were C16 : 0, summed feature 3 (C16 : 1 ω7c/C16 : 1 ω6c), summed feature 8 (C18 : 1 ω7c/C18 : 1 ω6c), and C17 : 0 cyclo. Phylogenetic analysis using the rpoD gene sequences and phylogenomic analyses based on the whole genome sequences demonstrated that the strains are members of the Pseudomonas fluorescens subgroup, but form a monophyletic and robust clade separated from their relatives. Average nucleotide identity and digital DNA–DNA hybridisation analyses with the closely related Pseudomonas species corroborated their novel species status. The strains were differentiated from their relatives by phenotypic characteristics, pathogenicity towards cyclamen, cellular fatty acid composition, and whole-cell MALDI-TOF mass spectrometry profiles. Based on the phenotypic, chemotaxonomic, and genotypic data obtained, we conclude that the strains represent a novel Pseudomonas species, for which we propose the name Pseudomonas cyclaminis sp. nov.; the type strain is MAFF 301449T (=ICMP 23720T).


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