Rhodococcus Qingshengii Research Articles

Complete genome sequence of Rhodococcus qingshengii phage Perlina.

Rhodococcus phage Perlina is a novel phage isolated on Rhodococcus qingshengii S10. Perlina encodes 112 open reading frames with typical phage structural genes identified and 3 tRNAs (tRNA-Ile, tRNA-Met, and tRNA-Asn). Few close relatives can be identified at the nucleotide level, suggesting a new phage species.

Rhodococcus qingshengii GlMm1 as the Basis of a Biosensor for Determination of the Fungicide Carbendazim

The possible application of Rhоdococcus qingshengii strain GlMm1, isolated from a Dead Sea clay sample, as the basis of a biosensor for determining the benzimidazole fungicide carbendazim was investigated. High sensitivity of the biosensor under neutral pH and up to 500 mM NaCl at 2 to 160 μM carbendazim was maintained for up to 30 days.

Potential of the plant growth-promoting rhizobacterium Rhodococcus qingshengii LMR356 in mitigating lead stress impact on Sulla spinosissima L.

Mining-related lead (Pb) pollution of the soil poses serious hazards to ecosystems and living organisms, including humans. Improved heavy metal phytoremediation efficacy, achieved by using phytostabilizing plants assisted by plant-growth-promoting (PGP) microorganisms, has been presented as an effective strategy for remediating polluted soils. The objective of this research was to examine the response and potential of the plant-growth-promoting bacterium LMR356, a Rhodococcus qingshengii strain isolated from an abandoned mining soil, under lead stress conditions. Compared to non-contaminated culture media, the presence of lead induced a significant decrease in auxin production (from 21.17 to 2.65 μg mL-1) and phosphate solubilization (from 33.60 to 8.22 mg L-1), whereas other PGP traits increased drastically, such as 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity (from 38.17 to 71.37 nmol mg-1 h-1 α-ketobutyrate), siderophore production (from 69 to 83%), exopolysaccharide production (from 1952.28 to 3637.72 mg mL-1), biofilm formation, and motility. We, therefore, investigated the behavior of Sulla spinosissima L. in the presence or absence of this strain under a variety of experimental conditions. Under hydroponic conditions, Sulla plants showed endurance to varying lead concentrations (500-1000 μM). Inoculation of plants with Rhodococcus qingshengii strain LMR356 enhanced plant tolerance, as demonstrated by the increase in plant biomass (ranging from 14.41 to 79.12%) compared to non-inoculated Pb-stressed and non-stressed control plants. Antioxidant enzyme activities (increasing by -42.71 to 126.8%) and chlorophyll (383.33%) and carotenoid (613.04%) content were also augmented. In addition to its impact on plant lead tolerance, strain LMR356 showed a growth-promoting effect on Sulla plants when cultivated in sterilized non-contaminated sand. Parameters such as plant biomass (16.57%), chlorophyll (24.14%), and carotenoid (30%) contents, as well as ascorbate peroxidase (APX), peroxidase (POD), and catalase (CAT) activities, were all elevated compared to non-inoculated plants. Furthermore, when the same plant species was cultivated in highly polluted soil, inoculation increased plant biomass and improved its physiological properties. These findings demonstrate that LMR356 is a phytobeneficial bacterial strain capable of enhancing Sulla growth under normal conditions and improving its heavy metal tolerance in multi-polluted soils. Thus, it can be considered a promising biofertilizer candidate for growing Sulla spinosissima L. or other selected plants intended for application in restoration and stabilization initiatives aimed at reviving and safeguarding environmentally compromised and polluted soils after mining activities.

Rhodococcus qingshengii GlMm1 as the Basis for a Biosensor for Determination of the Fungicide Carbendazim

Rhodococcus qingshengii GlMm1 as the Basis for a Biosensor for Determination of the Fungicide Carbendazim

Draft genome sequences of Chryseobacterium sp. MHB01, Rhodococcus qingshengii MHB02, and Agrobacterium tumefaciens MHB03 isolated from superabsorbent polymer granules inoculated with an arbuscular mycorrhizal fungus.

Chryseobacterium sp. MHB01, Rhodococcus qingshengii MHB02, and Agrobacterium tumefaciens MHB03 were isolated from superabsorbent polymer granules cultured with an arbuscular mycorrhizal fungus. Whole-genome sequencing of these three strains revealed genome sizes of 4.57 Mb, 7.13 Mb, and 5.49 Mb with G + C contents of 36.9%, 62.5%, and 58.2%, respectively.

Ecological features of trace elements tolerant microbes isolated from sewage sludge of urban wastewater treatment plant

Worldwide wastewater treatment plants generate enormous amounts of sewage sludge, and their further disposal depends on the treatment technologies applied and spontaneously occurring microbiological processes. From different ages urban sewage sludge, 12 strains of bacteria with simultaneous tolerance to two or more trace elements: Co, Ni, Cu, Zn, Cd and Pb at concentration of 3-5 mmol were isolated and identified by PCR of target genes and Sanger sequencing methods. The isloated metal(loids) tolerant strains belong to the species, i.e., Serratia fonticola, Rhodococcus qingshengii, Pseudomonas fragi, Pseudomonas extremaustralis, Pseudomonas cedrina, Stenotrophomonas maltophilia, Serratia liquefaciens and Citrobacter freundii. The ecological features of the isolated strains were studied. The optimal growth temperatures for most strains was 15–30°C at pH range of 5–9, although some strains grew at 7°C (Pseudomonas fragi SS0-4, Serratia fonticola SS0-9 and Serratia fonticola SS12-11). Satisfactory growth of two strains (Serratia fonticola SS0-1and Citrobacter freundii SS60-12) was noted in an acidic medium at pH 4. Most of the strains grew in the NaCl concentration range of 1–5%. The isolated bacteria resistant to high concentrations of trace elements can be used for the effective mineralization of sewage sludge and for the decontamination of wastewater.

ABA-metabolizing bacteria and rhamnolipids as valuable allies for enhancing phytoremediation efficiency in heavy metal-contaminated soils

Microbial-assisted phytoremediation has great potential to improve the efficiency of phytoremediation in heavy metal (HM)-contaminated soils. In this study, the synergistic effects of rhamnolipids and the abscisic acid (ABA)-metabolizing bacterium Rhodococcus qingshengii on the phytoremediation efficiency of Indian mustard (Brassica juncea) in HM-contaminated soils were investigated. The Cd, Zn, and Pb contents in plants treated with a combination of rhamnolipids and R. qingshengii were 48.4–77.1 %, 14.6–40.4 %, and 16.1–20.0 % higher, respectively, than in those treated with R. qingshengii alone, and 42.8–59.2 %, 13.1–48.2 %, and 7.3–67.5 % higher, respectively, than in those treated with rhamnolipids alone. In addition, the bioconcentration factors of each metal were improved, and the biomass further increased by 36.6–65.7 % compared to that of single treatments. Pearson's correlation analysis showed that rhamnolipids and R. qingshengii enhanced the accumulation of HMs in B. juncea by activating the available forms of HMs in the soil and regulating the ABA and indole-3-acetic acid in plants, respectively. The structural equation model indicated that R. qingshengii had a larger path coefficient than rhamnolipids in terms of HM content and plant biomass, suggesting that R. qingshengii may have a greater contribution to promoting the extraction of HMs from the soil under synergistic conditions. In conclusion, the combination of rhamnolipids and R. qingshengii has great potential to enhance the phytoremediation efficiency of hyperaccumulating plants in HM-contaminated soils.

Mechanistic insights into sulfur source-driven physiological responses and metabolic reorganization in the fuel-biodesulfurizing Rhodococcus qingshengii IGTS8.

Comparative proteomics and untargeted metabolomics were combined to study the physiological and metabolic adaptations of Rhodococcus qingshengii IGTS8 under biodesulfurization conditions. After growth in a chemically defined medium with either dibenzothiophene (DBT) or MgSO4 as the sulfur source, many differentially produced proteins and metabolites associated with several metabolic and physiological processes were detected including the metabolism of carbohydrates, amino acids, lipids, nucleotides, vitamins, protein synthesis, transcriptional regulation, cell envelope biogenesis, and cell division. Increased production of the redox cofactor mycofactocin and associated proteins was one of the most striking adaptations under biodesulfurization conditions. While most central metabolic enzymes were less abundant in the presence of DBT, a key enzyme of the glyoxylate shunt, isocitrate lyase, was up to 26-fold more abundant. Several C1 metabolism and oligotrophy-related enzymes were significantly more abundant in the biodesulfurizing culture. R. qingshengii IGTS8 exhibited oligotrophic growth in liquid and solid media under carbon starvation. Moreover, the oligotrophic growth was faster on the solid medium in the presence of DBT compared to MgSO4 cultures. In the DBT culture, the cell envelope and phospholipids were remodeled, with lower levels of phosphatidylethanolamine and unsaturated and short-chain fatty acids being the most prominent changes. Biodesulfurization increased the biosynthesis of osmoprotectants (ectoine and mannosylglycerate) as well as glutamate and induced the stringent response. Our findings reveal highly diverse and overlapping stress responses that could protect the biodesulfurizing culture not only from the associated sulfate limitation but also from chemical, oxidative, and osmotic stress, allowing efficient resource management. IMPORTANCE Despite decades of research, a commercially viable bioprocess for fuel desulfurization has not been developed yet. This is mainly due to lack of knowledge of the physiology and metabolism of fuel-biodesulfurizing bacteria. Being a stressful condition, biodesulfurization could provoke several stress responses that are not understood. This is particularly important because a thorough understanding of the microbial stress response is essential for the development of environmentally friendly and industrially efficient microbial biocatalysts. Our comparative systems biology studies provide a mechanistic understanding of the biology of biodesulfurization, which is crucial for informed developments through the rational design of recombinant biodesulfurizers and optimization of the bioprocess conditions. Our findings enhance the understanding of the physiology, metabolism, and stress response not only in biodesulfurizing bacteria but also in rhodococci, a precious group of biotechnologically important bacteria.

Microaerobic degradation of crude oil and long chain alkanes by a new Rhodococcus strain from Gulf of Mexico

Bacterial degradation of crude oil is a promising strategy for reducing the concentration of hydrocarbons in contaminated environments. In the first part of this study, we report the enrichment of two bacterial consortia from deep sediments of the Gulf of Mexico with crude oil as the sole carbon and energy source. We conducted a comparative analysis of the bacterial community in the original sediment, assessing its diversity, and compared it to the enrichment observed after exposure to crude oil in defined cultures. The consortium exhibiting the highest hydrocarbon degradation was predominantly enriched with Rhodococcus (75%). Bacterial community analysis revealed the presence of other hydrocarbonoclastic members in both consortia. In the second part, we report the isolation of the strain Rhodococcus sp. GOMB7 with crude oil as a unique carbon source under microaerobic conditions and its characterization. This strain demonstrated the ability to degrade long-chain alkanes, including eicosane, tetracosane, and octacosane. We named this new strain Rhodococcus qingshengii GOMB7. Genome analysis revealed the presence of several genes related to aromatic compound degradation, such as benA, benB, benC, catA, catB, and catC; and five alkB genes related to alkane degradation. Although members of the genus Rhodococcus are well known for their great metabolic versatility, including the aerobic degradation of recalcitrant organic compounds such as petroleum hydrocarbons, this is the first report of a novel strain of Rhodococcus capable of degrading long-chain alkanes under microaerobic conditions. The potential of R. qingshengii GOMB7 for applications in bioreactors or controlled systems with low oxygen levels offers an energy-efficient approach for treating crude oil-contaminated water and sediments.

Eco-friendly Biodegradation of Hydrocarbons Compounds from Crude Oily Wastewater Using PVA/Alginate/Clay Composite Hydrogels

Immobilized microorganisms especially bacteria are most used rather than free cells to be protected from the environmental conditions when being used for the bioremediation of environmental pollutants. Herein, two marine’s bacterial isolates were tested for their ability to decompose crude oil. The optimum conditions for effective bacterial degradation e.g., pH, temperature, and inoculum size were investigated. PVA-alginate-clay composite hydrogel beads with different types of incorporated mineral clays were prepared and tested as bacterial carrier for potential bioremediation. Synthesized composite hydrogels were physico-chemically characterized by FTIR, SEM, and thermal analyses. Results showed that, embedded degrading bacteria in PVA-alginate beads recorded degradation rates as 74 and 66.6% for both tested bacterial isolates (S and R) compared to 61.2 and 53% degradation rates by free cells, respectively. Where, attapulgite clay-containing beads recorded maximum degradation% as 78.8 and 75% for both bacterial isolates, when added to immobilization matrices and these percentages could be enhanced under optimal conditions. The 16S rRNA gene of the two marine oils degrading bacterial isolates were amplified and sequenced, where both isolates were identified as Pseudomonas stutzeri and Rhodococcus qingshengii with submitted accession numbers of ON908963 and ON908962, respectively. These results are referring to the ability of using both tested isolates for crude oil bioremediation process and embedded them into PVA-alginate-clay beads as hydrogel carrier under the optimum conditions.

Enhanced biodegradation of crude oil contamination by indigenous bacterial consortium under real conditions

This study was performed to evaluate the efficiency of using locally isolated bacteria to degrade crude oil in sea water-based medium. The bacteria were isolated from water samples taken from different sea environments (Mediterranean, Aegean, Marmara, and Black Sea). The experiments for crude oil degradation were carried out in small volume (100 mL sea water medium in 250 mL shaking flasks) or large volume (14 L sea water-based medium in a plastic container). Furthermore, the media were not sterilized and remained open to the environment (non-sterile conditions). On the other hand, the media were shaken at a low speed of 50 rpm to mimic sea conditions. Of twelve bacterial isolates, four isolates (PB1, PB4, PB6, and PB7) having higher oil degradation potential were selected. According to 16S rRNA analysis, the isolates PB4 and PB6 were identified as, respectively, Rhodococcus qingshengii and Alcanivorax venustensis, and the other two isolates (PB1 and PB7) as Pseudomonas sabulinigri. When the usability of four isolates alone or as a co-culture for crude oil degradation was tested, co-cultures were found to cause more crude oil degradation than individual bacterial cultures. Among the co-cultures, the maximum degradation of crude oil was achieved with the association of PB6 + PB4. During the experiments, carbon sources and minerals were not supplemented to the sea water medium, and even the supplementation of only nitrogen source (especially ammonium sulphate) was determined to significantly enhance the crude oil degradation potential of the bacterial consortium (PB6 + PB4). Spectrophotometric analyses (OD225) demonstrated that crude oil degradation continued up to 12th day in small volume but 16th day in large volume. GC-MS analyses revealed that co-culture of PB6 and PB4 degraded 100% of C9–C12 and C16–C29 hydrocarbons and 85% of C13–C15 hydrocarbons.

N fertilizers promote abscisic acid-catabolizing bacteria to enhance heavy metal phytoremediation from metalliferous soils

Bacterial-assisted phytoextraction is an attractive strategy to enhance the phytoremediation efficiency of heavy metal (HM)-contaminated soils. In the present study, we investigated the synergistic effects of N fertilizers and abscisic acid (ABA)-catabolizing bacteria on the HM (Cd, Zn, and Pb) phytoremediation efficiency of Brassica juncea L. (B. juncea). Compared with Rhodococcus qingshengii (R. qingshengii) alone, urea, ammonium‑nitrogen (NH4+-N), and nitrate‑nitrogen (NO3−-N) fertilizers combined with R. qingshengii increased HM concentrations in B. juncea by 13.8 %–48.2 %, 44.2 %–54.4 %, and 59.4 %–113.6 %, respectively, and enhanced the biomass of B. juncea by 7.7 %–38.8 %, 10.9 %–29.5 %, and 19.9 %–46.8 %, respectively. Consequently, the bioconcentration factor increased by 3.4 %–30.9 % and the phytoextraction rate increased by 18.5 %–98.7 %. Treatment with NO3−-N showed the most significant effect. In structural equation modeling, R. qingshengii inoculation showed greater path coefficients with soil pH and ABA and indoleacetic acid concentrations of B. juncea than N fertilization, indicating that R. qingshengii contributed more to HM extraction efficiency than N fertilizers. Additionally, differences in the extraction rates of Cd, Zn, and Pb from B. juncea were reduced following N fertilization. In summary, synergistic R. qingshengii inoculation and N fertilization have substantial potential to enhance phytoremediation efficiency. Combined application of R. qingshengii and NO3−-N fertilizers is recommended.

Detection of gene clusters for biodegradation of alkanes and aromatic compounds in the Rhodococcus qingshengii VKM Ac-2784D genome.

Bacterial species of the genus Rhodococcus are known to be efficient degraders of hydrocarbons in contaminated soil. They are also employed for bioremediation of polluted environments. These bacteria are widely met in soil, water and living organisms. Previously, we have isolated the Rhodococcus qingshengii strain VKM Ac-2784D from the rhizosphere of couch grass growing on oil-contaminated soil. This strain can effectively degrade oil and some model compounds (naphthalene, anthracene and phenanthrene). The results of phylogenetic analysis show that this strain belongs to the species R. qingshengii. To understand the catabolic properties of this strain, we have studied its gene clusters possessing such properties. The alkane destruction genes are represented by two clusters and five separate alkB genes. The destruction of aromatic compounds involves two stages, namely central and peripheral. The R. qingshengii VKM Ac-2784D genome contains four out of eight known central metabolic pathways for the destruction of aromatic compounds. The structure of the gene clusters is similar to that of the known strains R. jostii RHA1 and R. ruber Chol-4. The peripheral pathways include the genes encoding proteins for benzoic acid destruction. The presence of biphenyl 2,3-dioxygeneses as well as gene clusters of benzoate and 2-hydroxypentandienoate pathways suggests that R. qingshengii VKM Ac-2784D could degrade polychlorinated biphenyls. The biodegradation ability can be enhanced by biosurfactants, which are known to be synthesized by Rhodococcus. The R. qingshengii VKM Ac-2784D genome contains the otsA, otsB, treY, treZ genes. The bioinformatics data are supported by the previous biochemical experiments that allow a mixture of species with a wide variation of metabolic pathways to be obtained.

Design of novel water-soluble isoxazole-based antimicrobial agents and evaluation of their cytotoxicity and acute toxicity

Based on the readily available 3-organyl-5-(chloromethyl)isoxazoles, a number of previously unknown water-soluble conjugates of isoxazoles with thiourea, amino acids, some secondary and tertiary amines, and thioglycolic acid were synthesized. The bacteriostatic activity of aforementioned compounds has been studied against Enterococcus durans B-603, Bacillus subtilis B-407, Rhodococcus qingshengii Ac-2784D, and Escherichia coli B-1238 microorganisms (provided by All-Russian Collection of Microorganisms, VKM). The influence of the nature of the substituents in positions 3 and 5 of the isoxazole ring on the antimicrobial activity of the obtained compounds has been determined. It is found that the highest bacteriostatic effect is observed for compounds containing 4-methoxyphenyl or 5-nitrofuran-2-yl substituents in position 3 of the isoxazole ring as well as methylene group in position 5 bearing residues of l-proline or N-Ac-l-cysteine (5a–d, MIC 0.06–2.5 µg/ml). The leading compounds showed low cytotoxicity on normal human skin fibroblast cells (NAF1nor) and low acute toxicity on mice in comparison with the well-known isoxazole-containing antibiotic oxacillin.

Medium composition overturns the widely accepted sulfate-dependent repression of desulfurization phenotype in Rhodococcus qingshengii IGTS8.

Microbial desulfurization has been extensively studied as a promising alternative to the widely applied chemical desulfurization process. Sulfur removal from petroleum and its products becomes essential, as the environmental regulations become increasingly stringent. Rhodococcus qingshengii IGTS8 has gained ground as a naturally occurring model biocatalyst, due to its superior specific activity for desulfurization of dibenzothiophene (DBT). Recalcitrant organic sulfur compounds-DBT included-are preferentially removed by selective carbon-sulfur bond cleavage to avoid a reduction in the calorific value of the fuel. The process, however, still has not reached economically sustainable levels, as certain limitations have been identified. One of those bottlenecks is the repression of catalytic activity caused by ubiquitous sulfur sources such as inorganic sulfate, methionine, or cysteine. Herein, we report an optimized culture medium for wild-type stain IGTS8 that completely alleviates the sulfate-mediated repression of biodesulfurization activity without modification of the natural biocatalyst. Medium C not only promotes growth in the presence of several sulfur sources, including DBT, but also enhances biodesulfurization of resting cells grown in the presence of up to 5 mM sulfate. Based on the above, the present work can be considered as a step towards the development of a more viable commercial biodesulfurization process.

Efficacy of phytohormones producing isolates on the growth of two stem-cutting robusta coffee (Coffea canephora)

The propagation techniques of robusta coffee by cutting can be accelerated by the application of plant growth-promoting hormones. This study is to determine the efficacy of phytohormone-producing isolates on the growth of two-segment cutting of Robusta coffee. The study was conducted using a non-factorial completely randomized design with 2 replications. The treatments consist of control, synthetic IBA (Rooton-f) with three dilution concentrations, 4 isolates (Rhodococcus qingshengii, Burkholderia ambifaria, and Bacillus wiedmannii; fungi Aspergillus niger) with three dilution concentrations. Observed parameters included a percentage of live cuttings, percentage of cuttings sprouted, percentage of rooted cuttings, plant height, number of shoots, number of internodes, number of leaves, number of primary roots, primary root length, and primary root diameter. The results showed that the B. wiedmannii treatment had the best performance compared to other treatments because this isolate produced the highest number of primary roots (4.60) and primary root diameters (2.97 mm) compared to other treatments including control and Rooton-f. This is related to its ability to produce phytohormones that can stimulate the root growth of coffee robusta cuttings.

Low-temperature phenol-degrading microbial agent: construction and mechanism.

In this study, three cold-tolerant phenol-degrading strains, Pseudomonas veronii Ju-A1 (Ju-A1), Leifsonia naganoensis Ju-A4 (Ju-A4), and Rhodococcus qingshengii Ju-A6 (Ju-A6), were isolated. All three strains can produce cis, cis-muconic acid by ortho-cleavage of catechol at 12℃. Response surface methodology (RSM) was used to optimize the proportional composition of low-temperature phenol-degrading microbiota. Degradation of phenol below 160mgL-1 by low-temperature phenol-degrading microbiota followed first-order degradation kinetics. When the phenol concentration was greater than 200mgL-1, the overall degradation trend was in accordance with the modified Gompertz model. The experiments showed that the microbial agent (three strains of low-temperature phenol-degrading bacteria were fermented separately and constructed in the optimal ratio) could completely degrade 200mgL-1 phenol within 36h. The above construction method is more advantageous in bio-enhanced treatment of actual wastewater. Through the construction of microbial agents to enhance the degradation effect of phenol, it provides a feasible scheme for the biodegradation of phenol wastewater at low temperature and shows good application potential.

Complete Genome Sequence of Rhodococcus qingshengii Strain PM1, Isolated from a Selenium-Rich Mine in China.

Rhodococcus qingshengii PM1 was isolated from selenium-rich carbonaceous mudstones in Enshi, Hubei, China. Here, we report the complete genome sequence of this strain, which was obtained by combining Illumina and Nanopore sequencing.

Advancing Desulfurization in the Model Biocatalyst Rhodococcus qingshengii IGTS8 via an In Locus Combinatorial Approach.

Biodesulfurization poses as an ideal replacement to the high cost hydrodesulfurization of the recalcitrant heterocyclic sulfur compounds, such as dibenzothiophene (DBT) and its derivatives. The increasingly stringent limits on fuel sulfur content intensify the need for improved desulfurization biocatalysts, without sacrificing the calorific value of the fuel. Selective sulfur removal in a wide range of biodesulfurization strains, as well as in the model biocatalyst Rhodococcus qingshengii IGTS8, occurs via the 4S metabolic pathway that involves the dszABC operon, which encodes enzymes that catalyze the generation of 2-hydroxybiphenyl and sulfite from DBT. Here, using a homologous recombination process, we generate two recombinant IGTS8 biocatalysts, harboring native or rearranged, nonrepressible desulfurization operons, within the native dsz locus. The alleviation of sulfate-, methionine-, and cysteine-mediated dsz repression is achieved through the exchange of the native promoter Pdsz, with the nonrepressible Pkap1 promoter. The Dsz-mediated desulfurization from DBT was monitored at three growth phases, through HPLC analysis of end product levels. Notably, an 86-fold enhancement of desulfurization activity was documented in the presence of selected repressive sulfur sources for the recombinant biocatalyst harboring a combination of three targeted genetic modifications, namely, a dsz operon rearrangement, a native promoter exchange, and a dszA-dszB overlap removal. In addition, transcript level comparison highlighted the diverse effects of our genetic engineering approaches on dsz mRNA ratios and revealed a gene-specific differential increase in mRNA levels. IMPORTANCE Rhodococcus is perhaps the most promising biodesulfurization genus and is able to withstand the harsh process conditions of a biphasic biodesulfurization process. In the present work, we constructed an advanced biocatalyst harboring a combination of three genetic modifications, namely, an operon rearrangement, a promoter exchange, and a gene overlap removal. Our homologous recombination approach generated stable biocatalysts that do not require antibiotic addition, while harboring nonrepressible desulfurization operons that present very high biodesulfurization activities and are produced in simple and low-cost media. In addition, transcript level quantification validated the effects of our genetic engineering approaches on recombinant strains' dsz mRNA ratios and revealed a gene-specific differential increase in mRNA levels. Based on these findings, the present work can pave the way for further strain and process optimization studies that could eventually lead to an economically viable biodesulfurization process.

The Effect of Atranes on the Growth of <i>Rhodococcus qingshengii</i> VKM Ac-2784D in the Presence of Various Carbon Sources and on Its Ability to Degrade Naphthalene

Abstract—Rhodococcus qingshengii VKM Ac-2784D isolated from the rhizosphere of Elytrigia repens, is a promising oil degrader. To increase its biotechnological potential, the effect of atranes (potential growth stimulants) on its growth and naphthalene degradation was studied. The effect of the studied atranes on R. qingshengii VKM Ac-2784D was shown to depend on the nature of the carbon source: addition of atranes had practically no effect on the growth rate of bacteria in the rich BTN medium; in the media 8E and 8E with glucose, it baused short-term activation of growth, characterized by a shorter lag phase. In the presence of naphthalene, atranes had a predominantly negative effect on the growth of R. qingshengii VKM Ac-2784D. The studied atranes had a negative effect on the rate of naphthalene decomposition by R. qingshengii VKM Ac-2784D, and also decreased the cell wall hydrophobicity and the synthesis of extracellular biosurfactants. It may therefore be assumed that the studied compounds will have a negative effect on the rate of naphthalene decomposition by this microorganism.