A consortium of immobilized rhodococci for oilfield wastewater treatment in a column bioreactor

The possible use of a consortium of actinobacteria from the genus Rhodococcus immobilized on a polymeric carrier has been investigated for oilfield wastewater treatment in a bioreactor. It has been found that Rhodococcus opacus IEGM 263 and Rhodococcus ruber IEGM 231 cells remain viable at high concentrations of mineral salts in water and are able to oxidize oil hydrocarbons up to 62–81%. It has been shown that the consortium of rhodococci was more efficient in the elimination of hydrocarbons from wastewater than monocultures.     

Kinetics of the degradation of aliphatic hydrocarbons by the bacteria Rhodococcus ruber and Rhodococcus erythropolis

Consumption of aliphatic hydrocarbons by the bacteria Rhodococcus ruber Ac-1513-D and Rhodococcus erythropolis Ac-1514-D grown on mixed n-alkanes and diesel fuel was studied. Consumption of diesel fuel hydrocarbons by the strains was less intense in comparison with the n-alkane mixture. The strains showed differences in growth rate and consumption of the substrates, which suggests that they possess different mechanisms of hydrocarbon uptake.     

Relationships between Colony Morphotypes and Oil Tolerance in Rhodococcus rhodochrous

A mucoidal strain of Rhodococcus rhodochrous was resistant to 10% (vol/vol) n-hexadecane, while its rough derivatives were sensitive. When the extracellular polysaccharide (EPS) produced by the mucoidal strain was added to cultures of the rough strains, the rough strains gained resistance to n-hexadecane. Thus, EPS confer tolerance to n-hexadecane in members of the genus Rhodococcus.     

Biodesulfurization of dibenzothiophene and gas oil using a bioreactor containing a catalytic bed with Rhodococcus rhodochrous immobilized on silica

Biodesulfurization (BDS) in a bioreactor packed with a catalytic bed of silica containing immobilized Rhodococcus rhodochrous was studied. Various bed lengths and support particle sizes were evaluated for BDS of dibenzothiophene (DBT) and gas oil. The sulfur-containing substrates were introduced separately into the bioreactor at different feed flows. Higher removal of sulfur from DBT and gas oil was achieved with a long bed, lower substrate flow, and larger sizes of immobilization particles. The packed bed bioreactor containing metabolic active cells was recycled and maintained BDS activity.     

Synthesis of Imidazol-2-yl Amino Acids by Using Cells from Alkane-Oxidizing Bacteria

Sixty-one strains of alkane-oxidizing bacteria were tested for their ability to oxidize N-(2-hexylamino-4-phenylimidazol-1-yl)-acetamide to imidazol-2-yl amino acids applicable for pharmaceutical purposes. After growth with n-alkane, 15 strains formed different imidazol-2-yl amino acids identified by chemical structure analysis (mass and nuclear magnetic resonance spectrometry). High yields of imidazol-2-yl amino acids were produced by the strains Gordonia rubropertincta SBUG 105, Gordonia terrae SBUG 253, Nocardia asteroides SBUG 175, Rhodococcus erythropolis SBUG 251, and Rhodococcus erythropolis SBUG 254. Biotransformation occurred via oxidation of the alkyl side chain and produced 1-acetylamino-4-phenylimidazol-2-yl-6-aminohexanoic acid and the butanoic acid derivative. In addition, the acetylamino group of these products and of the substrate was transformed to an amino group. The product pattern as well as the transformation pathway of N-(2-hexylamino-4-phenylimidazol-1-yl)-acetamide differed in the various strains used.     

Biotransformations of 2-Methylisoborneol by Camphor-Degrading Bacteria

Many camphor-degrading bacteria that are able to transform 2-methylisoborneol (2-MIB) have been identified. Three of these strains have been examined in detail. Rhodococcus ruber T1 metabolizes camphor through 6-hydroxycamphor but converts 2-MIB to 3-hydroxy-2-MIB. Pseudomonas putida G1, which metabolizes camphor through 5-hydroxycamphor, converts MIB primarily to 6-hydroxy-2-MIB. Rhodococcus wratislaviensis DLC-cam converts 2-MIB through 5-hydroxy-2-MIB to 5-keto-2-MIB. Together, these three strains produce metabolites resulting from hydroxylation at all of the three available secondary carbons on the six-member ring of 2-MIB.     

喹啉废水反硝化反应器中优势菌的代谢功能分析

采用纯培养技术对一个降解喹啉的反硝化反应器筛选到的26株优势菌株进行反硝化能力以及好氧降解喹啉能力研究,其中的反硝化菌还测定了反硝化条件下的喹啉降解能力。结果发现Bacillus、Staphylococcus、Pseudomonas、Brucella、Delftia等5个属的10株菌具有反硝化能力,Rhodococcus属的9株细菌能够好氧降解喹啉,揭示了反硝化喹啉降解反应器中主要细菌类型的代谢特性,发现在缺氧反硝化反应器中存在多样的代谢类型的细菌。     

Cloning and characterization of a gene cluster for cyclododecanone oxidation in Rhodococcus ruber SC1

Biological oxidation of cyclic ketones normally results in formation of the corresponding dicarboxylic acids, which are further metabolized in the cell. Rhodococcus ruber strain SC1 was isolated from an industrial wastewater bioreactor that was able to utilize cyclododecanone as the sole carbon source. A reverse genetic approach was used to isolate a 10-kb gene cluster containing all genes required for oxidative conversion of cyclododecanone to 1,12-dodecanedioic acid (DDDA). The genes required for cyclododecanone oxidation were only marginally similar to the analogous genes for cyclohexanone oxidation. The biochemical function of the enzymes encoded on the 10-kb gene cluster, the flavin monooxygenase, the lactone hydrolase, the alcohol dehydrogenase, and the aldehyde dehydrogenase, was determined in Escherichia coli based on the ability to convert cyclododecanone. Recombinant E. coli strains grown in the presence of cyclododecanone accumulated lauryl lactone, 12-hydroxylauric acid, and/or DDDA depending on the genes cloned. The cyclododecanone monooxygenase is a type 1 Baeyer-Villiger flavin monooxygenase (FAD as cofactor) and exhibited substrate specificity towards long-chain cyclic ketones (C₁₁ to C₁₅), which is different from the specificity of cyclohexanone monooxygenase favoring short-chain cyclic compounds (C₅ to C₇).     

Biofouling inhibition in MBR by Rhodococcus sp. BH4 isolated from real MBR plant

It has been reported that an indigenous quorum quenching bacterium, Rhodococcus sp. BH4, which was isolated from a real plant of membrane bioreactor (MBR) has promising potential to control biofouling in MBR. However, little is known about quorum quenching mechanisms by the strain BH4. In this study, various characteristics of strain BH4 were investigated to elucidate its behavior in more detail in the mixed liquor of MBR. The N-acyl homoserine lactone hydrolase (AHL–lactonase) gene of strain BH4 showed a high degree of identity to qsdA in Rhodococcus erythropolis W2. The LC-ESI-MS analysis of the degradation product by strain BH4 confirmed that it inactivated AHL activity by hydrolyzing the lactone bond of AHL. It degraded a wide range of N-acyl homoserine lactones (AHLs), but there was a large difference in the degradation rate of each AHL compared to other reported AHL–lactonase-producing strains belonging to Rhodococcus genus. Its quorum quenching activity was confirmed not only in the Luria-Bertani medium, but also in the synthetic wastewater. Furthermore, the amount of strain BH4 encapsulated in the vessel as well as the material of the vessel substantially affected the quorum quenching activity of strain BH4, which provides useful information, particularly for the biofouling control in a real MBR plant from an engineering point of view.     

Catalase activity of hydrocarbon-oxidizing bacteria

The dynamics of catalase activity of the hydrocarbon-oxidizing bacteria Gordona terrae, Rhodococcus rubropertinctus, and Rhodococcus erythropolis during petroleum product destruction has been studied. A direct relationship between decreasing catalase activity of hydrocarbon-oxidizing microorganisms and the intensity of petroleum product destruction has been established experimentally. The revealed dependence allows one to consider the catalase activity of bacteria as an indicator of the initial stage of petroleum product oxidation and may be used for choosing destructor strains to construct biopreparations suitable for natural ecosystem remediation.     

Degradation of Estrogens by Rhodococcus zopfii and Rhodococcus equi Isolates from Activated Sludge in Wastewater Treatment Plants

We have isolated four strains of Rhodococcus which specifically degrade estrogens by using enrichment culture of activated sludge from wastewater treatment plants. Strain Y 50158, identified as Rhodococcus zopfii, completely and rapidly degraded 100 mg of 17β-estradiol, estrone, estriol, and ethinyl estradiol/liter, as demonstrated by thin-layer chromatography and gas chromatography-mass spectrometry analyses. Strains Y 50155, Y 50156, and Y 50157, identified as Rhodococcus equi, showed degradation activities comparable with that of Y 50158. Using the random amplified polymorphism DNA fingerprinting test, these three strains were confirmed to have been derived from different sources. R. zopfii Y 50158, which showed the highest activity among these four strains, revealed that the strain selectively degraded 17β-estradiol during jar fermentation, even when glucose was used as a readily utilizable carbon source in the culture medium. Measurement of estrogenic activities with human breast cancer-derived MVLN cells showed that these four strains each degraded 100 mg of 17β-estradiol/liter to 1/100 of the specific activity level after 24 h. It is thus suggested that these strains degrade 17β-estradiol into substances without estrogenic activity.     

Effect of bacterial satellites on Chlamydomonas reinhardtii growth in an algo-bacterial community

The growth characteristics of an algo-bacterial community (Chlamydomonas reinhardtii and bacterial satellites) were studied, as well as the mechanism and patterns of bacterial effect on algae. Four strains of predominant bacteria were isolated and partially characterized. They were assigned to the following taxa: Rhodococcus terrea, Micrococcus roseus, and Bacillus spp. A pure culture of the alga under study was obtained by plating serial dilutions on agarized media. Within the algo-bacterial association, the alga had a higher growth rate (0.76 day^?1) and yield (60 μg chlorophyll/ml culture) than in pure cultures (0.4 day^?1 and 10 μg chlorophyll/ml culture, respectively). The viability of the algal cells within the association was retained longer than in pure culture. Among the isolated bacterial satellites, strains B1 and Y1, assigned to the species Rhodococcus terrae, had the highest stimulatory effect on algal growth. The culture liquid of bacteria incubated under the conditions not permitting growth stimulated algal growth; the culture liquid of actively growing bacteria had an opposite effect.     

Screening of plant growth-promoting traits in arsenic-resistant bacteria isolated from the rhizosphere of soybean plants from Argentinean agricultural soil

Aims

The purpose of this study was to investigate plant-growth promoting traits in native and arsenic (As) highly-resistant bacterial strains isolated from the rhizosphere of soybean (Glycine max) plants grown in an Argentinean agricultural field.

Methods

Determination of MICs (Minimum inhibitory concentration) was carried out on solid media supplemented with arsenite (As 3+) or arsenate (As 5+). Morphological, cultural, physiological, biochemical and molecular characterization, and in vitro determination of plant growth promoting (PGP) properties of As resistant isolates were carried out. Arsenic in soil samples was determined by ICP-OES while residual arsenic on post-removal culture medium and accumulation in cells were estimated by GF-AAS after wet acid digestion.

Results

Isolated strains included γ-proteobacteria such as Enterobacter sp. and Pseudomonas sp., and actinobacteria as Rhodococcus sp. All bacterial strains grew in presence of very high arsenite -over 24?mM- and arsenate –over 400?mM- concentrations. Pseudomonas sp. strains presented simultaneously several in vitro PGP traits, although Rhodococcus erythropolis AW3 did not display PGP traits. However, R. erythropolis AW3 was the most As resistant strain and removed and accumulated the greatest amounts of the metalloid.

Conclusion

The presence of As resistant and plant-growth promoting bacterial strains in the rhizosphere of Glycine max, in arsenic containing agricultural soil, suggest that they could potentially play an important role in plant-growth promotion in stressed conditions. These strains were able to remove and accumulate As from liquid media, thus they could be beneficial for sustainable crop production.     

Cesium Accumulation and Growth Characteristics of Rhodococcus erythropolis CS98 and Rhodococcus sp. Strain CS402

Growth and cesium accumulation characteristics of two cesium-accumulating bacteria isolated from soils were investigated. Rhodococcus erythropolis CS98 and Rhodococcus sp. strain CS402 accumulated high levels of cesium (approximately 690 and 380 μmol/g [dry weight] of cells or 92 and 52 mg/g [dry weight] of cells, respectively) after 24 h of incubation in the presence of 0.5 mM cesium. The optimum pH for cesium uptake by both Rhodococcus strains was 8.5. Rubidium and cesium assumed part of the role of potassium in the growth of both Rhodococcus strains. Potassium and rubidium inhibited cesium accumulation by these Rhodococcus strains. It is likely that both Rhodococcus strains accumulated cesium through a potassium transport system.     

Acetylene degradation by new isolates of aerobic bacteria and comparison of acetylene hydratase enzymes

Aerobic acetylene-degrading bacteria were isolated from soil samples. Two isolates were assigned to the species Rhodococcus opacus, two others to Rhodococcus ruber and Gordona sp. They were compared with known strains of aerobic acetylene-, cyanide-, or nitrile-utilizing bacteria. The acetylene hydratases of R. opacus could be measured in cell-free extracts only in the presence of a strong reductant like titanium(III) citrate. Expression of these enzymes was molybdenum-dependent. Acetylene hydratases in cell-free extracts of R. ruber and Gordona spp. did not require addition of reductants. No cross-reactivity could be found between cell-free extracts of any of these aerobic isolates and antibodies raised against the acetylene hydratase of the strictly anaerobic fermenting bacterium Pelobacter acetylenicus. These results show that acetylene hydratases are a biochemically heterogeneous group of enzymes.     

Characterization of hydrocarbon-degrading bacterial strains isolated from oil-polluted soil

Using enrichment culturing method, a microbial population was detected in an oil-contaminated soil nearby an extraction field. Isolated strains were able use medium-length n-alkanes as sole carbon and energy source as assessed by growth experiments. Results showed a high diversity among strains at molecular level, and also in the metabolic profiles. Physiological and biochemical tests showed a similarity within a group of four strains, as confirmed by Biolog MicroLog analysis. Based on 16S rDNA sequences strains were identified as Rhodococcus erythropolis, Acinetobacter baumanii, Burkholderia cepacia and Achromobacter xylosoxidans. Alkane monooxygenase gene (alkB) was successfully detected in all our strains and for the first time alkB genes were found in an A. xylosoxidans strain. This bacterial species has been previously reported as part of microbial communities from oil polluted environments, but there are few studies that address mechanisms details of A. xylosoxidans involvement in n-alkane degradation process.     

Use of a Packed-Column Bioreactor for Isolation of Diverse Protease-Producing Bacteria from Antarctic Soil

Seventy-five aerobic heterotrophs have been isolated from a packed-column bioreactor inoculated with soil from Antarctica. The column was maintained at 10°C and continuously fed with a casein-containing medium to enrich protease producers. Twenty-eight isolates were selected for further characterization on the basis of morphology and production of clearing zones on skim milk plates. Phenotypic tests indicated that the strains were mainly psychrotrophs and presented a high morphological and metabolical diversity. The extracellular protease activities tested were optimal at neutral pH and between 30 and 45°C. 16S ribosomal DNA sequence analyses showed that the bioreactor was colonized by a wide variety of taxons, belonging to various bacterial divisions: α-, β-, and γ-Proteobacteria; the Flexibacter-Cytophaga-Bacteroides group; and high G+C gram-positive bacteria and low G+C gram-positive bacteria. Some strains represent candidates for new species of the genera Chryseobacterium and Massilia. This diversity demonstrates that the bioreactor is an efficient enrichment tool compared to traditional isolation strategies.     

Engineering Pseudomonas putida for efficient aromatic conversion to bioproduct using high throughput screening in a bioreactor

Pseudomonas putida KT2440 is an emerging biomanufacturing host amenable for use with renewable carbon streams including aromatics such as para-coumarate. We used a pooled transposon library disrupting nearly all (4,778) non-essential genes to characterize this microbe under common stirred-tank bioreactor parameters with quantitative fitness assays. Assessing differential fitness values by monitoring changes in mutant strain abundance identified 33 gene mutants with improved fitness across multiple stirred-tank bioreactor formats. Twenty-one deletion strains from this subset were reconstructed, including GacA, a regulator, TtgB, an ABC transporter, and PP_0063, a lipid A acyltransferase. Thirteen deletion strains with roles in varying cellular functions were evaluated for conversion of para-coumarate, to a heterologous bioproduct, indigoidine. Several mutants, such as the ΔgacA strain improved fitness in a bioreactor by 35 fold and showed an 8-fold improvement in indigoidine production (4.5 g/L, 0.29 g/g, 23% of maximum theoretical yield) from para-coumarate as the carbon source.     

The properties of hydrocarbon-oxidizing bacteria isolated from the oilfields of Tatarstan, western Siberia, and Vietnam

Eleven strains of hydrocarbon-oxidizing bacteria, isolated from oilfields and representing the genera Rhodococcus, Gordonia, Dietzia, and Pseudomonas, were characterized as mesophiles and neutrophiles. Rhodococci were halotolerant microorganisms growing in a media containing up to 15% NaCl. All the strains oxidized n-alkanes of crude oil. An influence of the cultivation temperatures (28 or 45°C) and organic supplements on the degradation of C₁₂-C₃₀ n-alkanes in oxidized oil by two bacterial strains of the genus Pseudomonas was shown. The introduction of acetate, propionate, butyrate, ethanol, and sucrose led mainly to decreased oxidation of petroleum paraffins. At certain cultivation temperatures, the addition of volatile fatty acid salts increased the content of certain n-alkanes in oxidized oil as compared to crude oil.