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1.
Isolation of a malachite green-degrading Pseudomonas sp. MDB-1 strain and cloning of the tmr2 gene 总被引:1,自引:0,他引:1
Lian-tai Li Qing Hong Xin Yan Gui-hua Fang Shinawar Waseem Ali Shun-peng Li 《Biodegradation》2009,20(6):769-776
The release of malachite green, a commonly used triphenylmethane dye, into the environment is causing increasing concern due
to its toxicity, mutagenicity, and carcinogenicity. A bacterial strain that could degrade malachite green was isolated from
the water of an aquatic hatchery. It was identified as a Pseudomonas sp. based on the morphological, physiological, and biochemical characteristics, as well as the analysis of 16S rRNA gene
sequence and designated as MDB-1. This strain was capable of degrading both malachite green and leucomalachite green, as well
as other triphenylmethane dyes including Crystal Violet and Basic Fuchsin. The gene tmr2, encoding the triphenylmethane reductase from MDB-1, was cloned, sequenced and effectively expressed in E. coli. These results highlight the potential of this bacterium for the bioremediation of aquatic environments contaminated by malachite
green. 相似文献
2.
Huan-Mei Li Lian-Tai Yan Cai-Fang Sun Jin-Jin Yuan-Gao Qing Hong Li Shun-Peng 《World journal of microbiology & biotechnology》2011,27(6):1323-1329
A bacterial strain K9 capable of degrading malachite green was isolated from the sludge of the wastewater treatment system
of a chemical plant. It was identified preliminarily as Pseudomonas sp. Strain K9 was also able to degrade other triphenylmethane dyes, such as Crystal Violet and Basic Fuchsin. The gene tmr2, encoding the triphenylmethane reductase, was cloned from strain K9, and functionally expressed in E. coli. A 5946-bp DNA fragment including the tmr2 gene was cloned from the genomic DNA of strain K9 by chromosome walking. Its sequence analysis showed that tmr2 was associated with a typical mobile element ISPpu12 consisting of tnpA (encoding a transposase), lspA (encoding a lipoprotein signal peptidase) and orf1 (encoding a putative MerR family regulator), orf2 (encoding a CDF family heavy metal/H+ antiporter). This association was also found in another malachite green-degrading strain Pseudomonas sp. MDB-1, which indicated that the tmr2 gene might be a horizontally transferable gene. 相似文献
3.
Jang MS Lee YM Kim CH Lee JH Kang DW Kim SJ Lee YC 《Applied and environmental microbiology》2005,71(12):7955-7960
We purified to homogeneity an enzyme from Citrobacter sp. strain KCTC 18061P capable of decolorizing triphenylmethane dyes. The native form of the enzyme was identified as a homodimer with a subunit molecular mass of about 31 kDa. It catalyzes the NADH-dependent reduction of triphenylmethane dyes, with remarkable substrate specificity related to dye structure. Maximal enzyme activity occurred at pH 9.0 and 60 degrees C. The enzymatic reaction product of the triphenylmethane dye crystal violet was identified as its leuco form by UV-visible spectral changes and thin-layer chromatography. A gene encoding this enzyme was isolated based on its N-terminal and internal amino acid sequences. The nucleotide sequence of the gene has a single open reading frame encoding 287 amino acids with a predicted molecular mass of 30,954 Da. Although the deduced amino acid sequence displays 99% identity to the hypothetical protein from Listeria monocytogenes strain 4b H7858, it shows no overall functional similarity to any known protein in the public databases. At the N terminus, the amino acid sequence has high homology to sequences of NAD(P)H-dependent enzymes containing the dinucleotide-binding motif GXXGXXG. The enzyme was heterologously expressed in Escherichia coli, and the purified recombinant enzyme showed characteristics similar to those of the native enzyme. This is the first report of a triphenylmethane reductase characterized from any organism. 相似文献
4.
随着我国印染工业的发展,废水对生态环境的危害日趋严重,亟需开发一种脱色明显且成本低廉的降解方法。本研究发现毛木耳Auricularia cornea菌株AC5对不同结构的染料均具有一定的降解作用,尤其是三苯甲烷类染料。利用26℃、160r/min振荡培养7d的粗酶液对染料(75.0mg/L)进行12h降解,结果显示三苯甲烷染料孔雀石绿、结晶紫,蒽醌染料活性蓝19和偶氮染料活性蓝222的降解效率分别为83.27%、71.77%、67.81%和63.92%。染料降解实验和酶活力测定结果表明,毛木耳对孔雀石绿的降解率达到最高时漆酶活性最高,为321.0U/mL,木质素过氧化物酶和锰过氧化物酶活性较低。因此,推测在降解过程中漆酶起到主要作用。研究表明利用毛木耳菌丝发酵液降解染料废水成本低且操作方便,为染料废水的降解研究提供了前期基础。 相似文献
5.
Triphenylmethane Reductase from Citrobacter sp. Strain KCTC 18061P: Purification, Characterization, Gene Cloning, and Overexpression of a Functional Protein in Escherichia coli 下载免费PDF全文
Moon-Sun Jang Young-Mi Lee Cheorl-Ho Kim Jai-Heon Lee Dong-Woo Kang Seok-Jo Kim Young-Choon Lee 《Applied microbiology》2005,71(12):7955-7960
We purified to homogeneity an enzyme from Citrobacter sp. strain KCTC 18061P capable of decolorizing triphenylmethane dyes. The native form of the enzyme was identified as a homodimer with a subunit molecular mass of about 31 kDa. It catalyzes the NADH-dependent reduction of triphenylmethane dyes, with remarkable substrate specificity related to dye structure. Maximal enzyme activity occurred at pH 9.0 and 60°C. The enzymatic reaction product of the triphenylmethane dye crystal violet was identified as its leuco form by UV-visible spectral changes and thin-layer chromatography. A gene encoding this enzyme was isolated based on its N-terminal and internal amino acid sequences. The nucleotide sequence of the gene has a single open reading frame encoding 287 amino acids with a predicted molecular mass of 30,954 Da. Although the deduced amino acid sequence displays 99% identity to the hypothetical protein from Listeria monocytogenes strain 4b H7858, it shows no overall functional similarity to any known protein in the public databases. At the N terminus, the amino acid sequence has high homology to sequences of NAD(P)H-dependent enzymes containing the dinucleotide-binding motif GXXGXXG. The enzyme was heterologously expressed in Escherichia coli, and the purified recombinant enzyme showed characteristics similar to those of the native enzyme. This is the first report of a triphenylmethane reductase characterized from any organism. 相似文献
6.
A. Bajaj A. Pathak M.R. Mudiam S. Mayilraj N. Manickam 《Journal of applied microbiology》2010,109(6):2135-2143
Aim: To isolate bacteria capable of degrading endosulfan (ES) and the more toxic ES sulfate and to characterize their metabolites. Methods and Results: A Pseudomonas sp. strain IITR01 capable of degrading α‐ES and toxic ES sulfate was isolated using technical‐ES through enrichment culture techniques. No growth and no degradation were observed using β‐ES. Thin‐layer chromatography and gas chromatography‐mass spectrum analysis revealed the disappearance of both α‐ES and ES sulfate and the formation of hydroxylated products ES diol, ether and lactone. We show here for the first time the formation of aforementioned metabolites in contrast to ES hemisulfate yielded by an Arthrobacter sp. Metabolism of α‐ES and endosulfate was also observed using the crude cell extract of IITR01. The molecular mass of protein induced during the degradation of α‐ES and sulfate as substrate was found to be approximately 150 kDa as determined by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). Conclusion: We describe characterization of bacterium capable of degrading α‐ES and ES sulfate but not β‐ES. Genetic investigation suggests that a gene nonhomologous to the reported esd may be present in the strain IITR01. Significance and Impact of the Study: This study describes toxic ES degradation by a Pseudomonas species that may be utilized for the bioremediation of the industrial soils contaminated with ES residues. 相似文献
7.
Tao Pan Suizhou Ren Meiying Xu Guoping Sun Jun Guo 《Applied microbiology and biotechnology》2013,97(13):6051-6055
The biological treatment of triphenylmethane dyes is an important issue. Most microbes have limited practical application because they cannot completely detoxicate these dyes. In this study, the extractive biodecolorization of triphenylmethane dyes by Aeromonas hydrophila DN322p was carried out by introducing the cloud point system. The cloud point system is composed of a mixture of nonionic surfactants (20 g/L) Brij 30 and Tergitol TMN-3 in equal proportions. After the decolorization of crystal violet, a higher wet cell weight was obtained in the cloud point system than that of the control system. Based on the results of thin-layer chromatography, the residual crystal violet and its decolorized product, leuco crystal violet, preferred to partition into the coacervate phase. Therefore, the detoxification of the dilute phase was achieved, which indicated that the dilute phase could be discharged without causing dye pollution. The extractive biodecolorization of three other triphenylmethane dyes was also examined in this system. The decolorization of malachite green and brilliant green was similar to that of crystal violet. Only ethyl violet achieved a poor decolorization rate because DN322p decolorized it via adsorption but did not convert it into its leuco form. This study provides potential application of biological treatment in triphenylmethane dye wastewater. 相似文献
8.
【目的】筛选海洋来源的多糖降解菌,分析其多糖降解能力并初探机制。【方法】碘液染色法从海泥中初筛琼脂糖降解菌,唯一碳源生长法分析菌株的多糖利用能力,克隆16S rRNA基因以分析系统分类地位。用硫酸铵沉淀法制备胞外粗酶制剂,DNS-还原糖法测定琼胶酶活性,活性染色法分析胞外琼胶酶系的组成特征。分离、纯化琼脂糖的酶解产物,通过TLC测定寡糖Rf值、阳离子质谱测定分子量。【结果】分离到1株能液化琼脂糖的海洋细菌JZB09,鉴定至桃色杆菌属(Persicobacter)。JZB09能利用11种不同的多糖为唯一碳源生长,在利用琼脂糖、纤维素和木聚糖时生长较好。胞外粗酶制剂的琼胶酶活力约77.2U/mg,含有至少2条琼胶酶,大小约45kDa、70kDa。酶制剂降解琼脂糖后的产物是系列新琼寡糖,四糖是主产物,表明β-琼胶酶在胞外琼胶酶系降解琼脂糖时起关键作用。【结论】海洋细菌Persicobacter sp.JZB09是1株多能型多糖降解菌,可分泌β-琼胶酶降解琼脂糖且活性显著,具有潜在开发价值。 相似文献
9.
A broad-spectrum dye-decolorizing bacterium, strain DN322, was isolated from activated sludge of a textile printing wastewater treatment plant. The strain was characterized and identified as a member of Aeromonas hydrophila based on Gram staining, morphology characters, biochemical tests, and nearly complete sequence analysis of 16S rRNA gene and the gyrase subunit beta gene (gyrB). Strain DN322 decolorized a variety of synthetic dyes, including triphenylmethane, azo, and anthraquinone dyes. For color removal, the most suitable pH and temperature were pH 5.0–10.0 and 25–37°C, respectively. Triphenylmethane dye, e.g., Crystal Violet, Basic Fuchsin, Brilliant Green, and Malachite Green (50 mg l−1) were decolorized more than 90% within 10 h under aerobic culture condition and Crystal Violet could be used as sole carbon source and energy source for cell growth. The color removal of triphenylmethane dyes was due to a soluble cytosolic enzyme, and the enzyme was an NADH/NADPH-dependent oxygenase; For azo and anthraquinone dyes, e.g., Acid Amaranth, Great Red GR, Reactive Red KE-3B, and Reactive Brilliant Blue K-GR (50 mg l−1) could be decolorized more than 85% within 36 h under anoxic condition. This strain may be useful for bioremediation applications. 相似文献
10.
A novel isolate of a bacterium, capable of degrading trichloroethylene (TCE) and growing on this as the sole carbon source
is reported. The test strain was isolated by an enrichment technique with trichloroethylene as the substrate. The isolated
strain belongs to the genus Bacillus. The practical utility of cleaning up oil spillage by bioremediation could be extended to this bacterium to degrade the environmental
pollutant, which is used in metal degreasing in industries. Cells of the novel bacterium immobilized on calcium alginate were
found to have better trichloroethylene degrading activity than the ones which were immobilized on agar-agar or free cells. 相似文献
11.
Identification and characterization of chlorpyrifos-methyl and 3,5,6-trichloro-2-pyridinol degrading Burkholderia sp. strain KR100 总被引:1,自引:0,他引:1
A chlorpyrifos-methyl (CM) degrading bacterium (designated strain KR100) was isolated from a Korean rice paddy soil and was
further tested for its sensitivity against eight commercial antibiotics. Based on morphological, biochemical, and molecular
characteristics, this bacterium showed greatest similarity to members of the order Burkholderiales and was shown to be most
closely related to members of the Burkholderia cepacia group. Strain KR100 hydrolyzed CM to 3,5,6-trichloro-2-pyridinol (TCP) and utilized TCP as the sole source of carbon for
its growth. The isolate was also able to degrade chlorpyrifos, dimethoate, fenitrothion, malathion, and monocrotophos at 300 μg/ml
but diazinon, dicrotophos, parathion, and parathion-methyl at 100 μg/ml. The ability to degrade CM was found to be encoded
on a single plasmid of ~50 kb, pKR1. Genes encoding resistance to amphotericin B, polymixin B sulfate, and tetracycline were
also located on the plasmid. This bacterium merits further study as a potential biological agent for the remediation of soil,
water, or crop contaminated with organophosphorus compounds because of its greater biodegradation activity and its broad specificity
against a range of organophosphorus insecticides. 相似文献
12.
Decolorization of industrial dyes by a Brazilian strain of Pleurotus pulmonarius producing laccase as the sole phenol-oxidizing enzyme 总被引:2,自引:0,他引:2
The ability of a Brazilian strain ofPleurotus pulmonarius to decolorize structurally different synthetic dyes (including azo, triphenylmethane, heterocyclic and polymeric dyes) was
investigated in solid and submerged cultures. Both were able to decolorize completely or partially 8 of 10 dyes (Amido Black,
Congo Red, Trypan Blue, Methyl Green, Remazol Brilliant Blue R, Methyl Violet, Ethyl Violet, Brilliant Cresyl Blue). No decolorization
of Methylene Blue and Poly R 478 was observed. Of the four phenol-oxidizing enzymes tested in culture filtrates (lignin peroxidase,
manganese peroxidase, aryl alcohol oxidase, laccase),P. pulmonarius produced only laccase. Both laccase activity and dye decolorization were related to glucose and ammonium starvation or to
induction by ferulic acid. The decolorizationin vivo was tested using three dyes — Remazol Brilliant Blue R, Trypan Blue and Methyl Green. All of them were completely decolorized
by crude extracellular extracts. Decolorization and laccase activity were equally affected by pH and temperature. Laccase
can thus be considered to be the major enzyme involved in the ability ofP. pulmonarius to decolorize industrial dyes. 相似文献
13.
Decolorization of textile dyes by enzymatic extract and submerged cultures of <Emphasis Type="Italic">Pleurotus sajor-caju</Emphasis> 总被引:1,自引:0,他引:1
Fernanda M. Munari Tamara A. Gaio Raquel Calloni Aldo J. P. Dillon 《World journal of microbiology & biotechnology》2008,24(8):1383-1392
Pleurotus sajor-caju PS2001 was screened in Petri dish plates to assess the dye-decolorizing ability of industrial textile dyes. P. sajor-caju PS2001 was also cultivated in solid-state fermentation containing sawdust of Pinus sp. and wheat bran to obtain the enzymatic extract, showing laccase and manganese-peroxidase activity, which was used to
test the capacity to degrade the textile dyes. Additional tests of decolorization were performed in liquid cultures. Anthraquinone-type
textile dyes proved to be substrates for the enzymatic system of P. sajor-caju PS2001. Cultures in Petri dish plates showed that the anthraquinone dye Reactive Blue 220 can act as a redox mediator for
the enzymatic reactions involved in the decolorization process, and enables the azo dye degradation. Reactive Blue 220 and
Acid Blue 280 were completely decolorized in 30 min and 60 min, respectively, during the tests with precipitated enzymatic
extract, while the azo dyes showed resistance to degradation. Additionally, in submerged cultures with dyes, veratryl alcohol
oxidases and lignin peroxidase activities were observed. These results suggest that the strain P. sajor-caju PS2001 has great potential for use in the bioremediation technology of recalcitrant pollutant such as textile effluents. 相似文献
14.
A pentachlorophenol (PCP) degrading bacterium was isolated and characterized from sludge of pulp and paper mill. This isolate
used PCP as its sole source of carbon and energy and was capable of degrading this compound, as indicated by stoichiometric
release of chloride and biomass formation. Based on morphology, biochemical tests, and 16S rRNA gene sequence analysis this
strain was identified as Kocuria sp. CL2. High Performance Liquid Chromatography (HPLC) analysis revealed that this strain was able to degrade PCP up to a
concentration of 600 mg/l. This is first time we are reporting the degradation of PCP by the Kocuria species. This isolate was also able to remove 58.64% of PCP from the sludge within two weeks. This study showed that the
removal efficiency of PCP by CL2 was found to be very effective and can be used in degradation of PCP containing pulp paper
mill waste in the environment. 相似文献
15.
Liu W Chao Y Yang X Bao H Qian S 《Journal of industrial microbiology & biotechnology》2004,31(3):127-132
One laccase-secreting engineered strain and four white-rot fungi were tested for their capacity to decolorize nine dyes that could be classified as azo, anthraquinonic and triphenylmethane dyes. Trametes versicolor was the most efficient of the tested strains under these experimental conditions. Anthraquinonic dyes were decolorized more easily than the other two types. Small structural differences among the dyes could significantly affect decolorization. None of the strains showed lignin peroxidase or veratryl alcohol oxidase activity. None of the dyes were decolorized completely by laccase alone. It is likely that other phenoloxidases, such as Mn-dependent and versatile peroxidase, were also involved in decolorization of the dyes. 相似文献
16.
Masuda M Yamasaki Y Ueno S Inoue A 《Extremophiles : life under extreme conditions》2007,11(2):355-362
Bisphenol A (BPA) is a highly biotoxic compound that kills many microorganisms at a low concentration (1,000 ppm). We isolated
BPA-tolerant/degrading Pseudomonas monteilii strain N-502 from about 1,000 samples collected from a field, sewage, and pond water. The isolated strain had strong BPA
tolerance and high BPA-degrading activity. This strain was able to grow in a minimum medium containing BPA as the sole carbon
source. Strain N-502 is an aerobic, motile, gram-negative, nonspore-forming, rod-shaped bacterium and was identified as P. monteilii, based on 16 S rRNA gene analysis. Strain N-502 completely degraded BPA 500 ppm in a 10-day, in culture system and was able
to degrade BPA 100 ppm in a 2-h resting cell system. This strain also showed potent ability to degrade BPA 500 and 1,000 ppm
in the resting cell system. Moreover, the initial BPA degradation rate was accelerated with the addition of Ca2+, Mg2+, and folic acid. 相似文献
17.
Koushik Mukherjee Prosun Tribedi Arup Chowdhury Tanusree Ray Archi Joardar Subhajit Giri Alok Kumar Sil 《Biodegradation》2011,22(2):377-388
Polyurethane diol (PUR-diol), a synthetic polymer, is widely used as a modifier for water-soluble resins and emulsions in
wood appliances and auto coatings. Non-biodegradability of polyurethanes (PUR) and PUR-based materials poses a threat to environment
that has led scientists to isolate microbes capable of degrading PUR. However, the bio-degradation of PUR-diol has not yet
been reported. In this study, we report isolation of a soil bacterium that can survive using PUR-diol as sole carbon source.
PUR-diol degradation by the organism was confirmed by thin layer chromatographic analysis of the conditioned medium obtained
after the growth wherein a significant reduction of PUR-diol was observed compared to non-inoculated medium. To quantify the
PUR-diol degradation, a sensitive assay based on High Performance Thin Layer Chromatography has been developed that showed
32% degradation of PUR-diol by the organism in 10 days. Degradation kinetics showed the maximal depletion of PUR-diol during
logarithmic growth of the organism indicating a direct relation between the growth and PUR-diol degradation. Mutagenic study
and GC-MS analysis revealed that esterase activity is involved in this degradation event. The ribotyping and metabolic fingerprinting
analysis showed that this organism is a strain of Pseudomonous aeruginosa (P. aeruginosa). It has also been observed that this strain is able to degrade Impranil DLN™, a variety of commercially available PUR. Therefore
this study identifies a new bacterium from soil that has the potential to reduce PUR-related waste burden and adds a new facet
to diverse functional activities of P. aeruginosa. 相似文献
18.
三氯乙烯降解菌FT17的分离、鉴定及其降解特性研究 总被引:2,自引:0,他引:2
采用水-硅油双相系统, 从辽河流域浑河沈阳段底泥中筛选得到一株三氯乙烯降解菌FT17。综合形态特征、生理生化特征、16S rRNA Blast分析和系统发育分析结果, 将该菌株鉴定为Sporosarcina ginsengisoli。菌株FT17最适生长温度为34°C, 最适生长pH为7.8。苯酚作为共代谢基质可以促进该菌株对三氯乙烯的降解。该菌株的三氯乙烯降解酶在胞内和胞外均存在。采用两种质粒提取方法对该菌株进行质粒检测, 结果均没有发现质粒条带, 推测该菌株的三氯乙烯降解基因位于染色体上。 相似文献
19.
A phenazine-1-carboxylic acid (PCA)–degrading bacterium, strain DP58, was isolated from pimiento rhizosoil. Based on morphology,
physiologic tests, 16S rDNA sequence, and phylogenetic characteristics, it was identified as Sphingomonas sp. The PCA-degradation experiments were conducted both in Luria-Bertani and inorganic salt medium at 28°C. The relationship
between bacterium growth and PCA degradation suggested that strain DP58 could use PCA as the sole source of carbon and nitrogen
and was able to completely degrade PCA in 40 hours. Newly isolated strain DP58 represents the first bacterium that can degrade
PCA. 相似文献
20.
Z-phenylacetaldoxime degradation by a novel aldoxime dehydratase from Bacillus sp. strain OxB-1 总被引:2,自引:0,他引:2
Z-phenylacetaldoxime (Z-PAOx) degrading bacterium, identified as Bacillus sp. strain OxB-1, was isolated from soil after 2 months acclimation. The enzyme involved in the degradation of Z-PAOx was induced by the aldoxime and required FMN for its activity. The enzyme was partially purified from the cell-free extract of the strain and shown to catalyze the stoichiometric dehydration reaction of Z-PAOx to form phenylacetonitrile (PAN). Activities of nitrilase and amidase acting on PAN and phenylacetamide (PAAm), respectively, to form phenylacetate (PAA) were found in the strain grown on Z-PAOx. This is the first report of aldoxime dehydratase co-existing with nitrile degrading enzymes in bacteria. 相似文献