Acinetobacter Sp Research Articles

Crude Oil Biodegradation by a Biosurfactant-Producing Bacterial Consortium in High-Salinity Soil

Bioremediation is a promising strategy to remove crude oil contaminants. However, limited studies explored the potential of bacterial consortia on crude oil biodegradation in high salinity soil. In this study, four halotolerant strains (Pseudoxanthomonas sp. S1-2, Bacillus sp. S2-A, Dietzia sp. CN-3, and Acinetobacter sp. HC8-3S), with strong environmental tolerance (temperature, pH, and salinity), distinctive crude oil degradation, and beneficial biosurfactant production, were combined to construct a bacterial consortium. The inoculation of the consortium successfully degraded 97.1% of total petroleum hydrocarbons in 10 days, with notable removal of alkanes, cycloalkanes, branched alkanes, and aromatic hydrocarbons. Functional optimization showed that this consortium degraded crude oil effectively in a broad range of temperature (20–37 °C), pH (6–9), and salinity (0–100 g/L). In salt-enriched crude-oil-contaminated soil microcosms, the simultaneous treatment of bioaugmentation and biostimulation achieved the highest crude oil degradation rate of 568.6 mg/kg/d, compared to treatments involving abiotic factors, natural attenuation, biostimulation, and bioaugmentation after 60 days. Real-time PCR targeting the 16S rRNA and alkB genes showed the good adaptability and stability of this consortium. The degradation property of the constructed bacterial consortium and the engineered consortium strategy may have potential use in the bioremediation of crude oil pollution in high-salinity soil.

Enhanced stereodivergent evolution of carboxylesterase for efficient kinetic resolution of near-symmetric esters through machine learning

Carboxylesterases serve as potent biocatalysts in the enantioselective synthesis of chiral carboxylic acids and esters. However, naturally occurring carboxylesterases exhibit limited enantioselectivity, particularly toward ethyl 3-cyclohexene-1-carboxylate (CHCE, S1), due to its nearly symmetric structure. While machine learning effectively expedites directed evolution, the lack of models for predicting the enantioselectivity for carboxylesterases has hindered progress, primarily due to challenges in obtaining high-quality training datasets. In this study, we devise a high-throughput method by coupling alcohol dehydrogenase to determine the apparent enantioselectivity of the carboxylesterase AcEst1 from Acinetobacter sp. JNU9335, generating a high-quality dataset. Leveraging seven features derived from biochemical considerations, we quantitively describe the steric, hydrophobic, hydrophilic, electrostatic, hydrogen bonding, and π-π interaction effects of residues within AcEst1. A robust gradient boosting regression tree model is trained to facilitate stereodivergent evolution, resulting in the enhanced enantioselectivity of AcEst1 toward S1. Through this approach, we successfully obtain two stereocomplementary variants, DR3 and DS6, demonstrating significantly increased and reversed enantioselectivity. Notably, DR3 and DS6 exhibit utility in the enantioselective hydrolysis of various symmetric esters. Comprehensive kinetic parameter analysis, molecular dynamics simulations, and QM/MM calculations offer insights into the kinetic and thermodynamic features underlying the manipulated enantioselectivity of DR3 and DS6.

Development of biocatalysts for high-value-added compounds.

High-value-added compounds, such as monoterpenoids, are an important industrial targets because they are an essential group of compounds for pharmaceutical and industrial applications. Meanwhile, the depletion of natural resources and climate change demands sustainable production methods. In recent years, biocatalysis, which allows microbial bioproduction by regio- and stereo-selective reaction under mild conditions, has been attracted researchers' attention as a possible alternative to conventional methods. In this mini-review, we focus on the identification of biotransformation pathways in the non-model microorganism Acinetobacter sp. Tol 5 using geraniol, a representative monoterpenoid, and on the construction of an unconventional bioproduction method for high-value-added monoterpenoid (E)-geranic acid, which has great potential for industrial applications. This method offers a more environmentally friendly approach and insights contribute to optimizing biotransformation and bioproduction strategies for high-value-added compounds.

Novel Acinetobacter sp. Isolated from Oil-contaminated Soil for Microbial Enhanced Oil Recovery

The viscosity of hydrocarbon reservoirs increases with age. So, a significant amount of oil is trapped in the underground reservoirs after recovery using primary and secondary methods. To improve the recovery, tertiary methods have been used. However, there is limited research on Microbial Enhanced Oil Recovery (MEOR) as a tertiary method due to microbes' inability to survive the high-temperature reservoir conditions. Consequently, the goal of this research is the creation of a novel bacteria strain for MEOR. The isolated bacteria colonies were inoculated in a batch fermentation broth, and the biosurfactant produced was screened using the oil displacement, emulsification index, modified drop-collapse test, surface tension, and interfacial tension criteria. The effects of pH (7.2 – 10.52), and salinity (15 – 35 %) at optimum temperature were studied on the selected isolate, which was identified by partial 16 rRNA gene sequence analysis. Gas Chromatography-Mass Spectroscopy was used to characterize the biosurfactants produced. The new isolate reduced heavy oil viscosity by 17% and produced a recovery factor in the range of 13-16.72%. The new bacteria is a thermophile and survived at a temperature of 65 °C, indicating promise for use in MEOR.

Combined high-rate contact stabilization and chemically enhanced primary treatment for enhanced recovery of organic matter and biogas from sewage

This study examined integrating high-rate contact stabilization (HRCS) and chemically enhanced primary treatment (CEPT) for wastewater to improve the carbon recovery rate (CRR). Enhancing chemical oxygen demand (COD) removal efficiency was hypothesized to improve the CRR. The evaluation covered serial HRCS-CEPT, serial CEPT-HRCS, and single-stage carbon recovery (Single-CR). The COD removal efficiencies for individual HRCS and CEPT were 50.3 % and 56.2 %, respectively. The serial CEPT-HRCS system failed in the HRCS process due to poor settling, resulting in microbial washout. However, the serial HRCS-CEPT system achieved the highest COD removal efficiency (84.5 %). The Single-CR system exhibited the highest CRR of 0.780 ± 0.083 g-CODCH4/g-CODinf, identifying it as the most promising process for energy-positive wastewater treatment. The selective pressure in the high-rate system resulted in a simplified and specialized bacterial community, mainly comprising microorganisms with high polyhydroxyalkanoate storage capacity, such as Lactococcus sp., Enterobacter sp., and Acinetobacter sp.

A tailored cytochrome P450 monooxygenase from Gordonia rubripertincta CWB2 for selective aliphatic monooxygenation.

Cytochrome P450 monooxygenases are recognized as versatile biocatalysts due to their broad reaction capabilities. One important reaction is the hydroxylation of non-activated C-H bonds. The subfamily CYP153A is known for terminal hydroxylation reactions, giving access to functionalized aliphatics. Whilst fatty derivatives may be converted by numerous enzyme classes, midchain aliphatics are seldomly accepted, a prime property of CYP153As. We report here on a new CYP153A member from the genome ofthe mesophilic actinobacterium Gordonia rubripertincta CWB2 as an efficient biocatalyst. The gene was overexpressed inEscherichia coli and fused with a surrogate electron transport system from Acinetobacter sp. OC4. This chimeric self-sufficient whole-cell system could perform hydroxylation and epoxidation reactions: conversions of C6-C14 alkanes, alkenes, alcohols and of cyclic compounds were observed, yielding production rates of, e.g., 2.69 mM h-1 for 1-hexanol and 4.97 mM h-1 for 1,2-epoxyhexane. Optimizing the linker compositions between the protein units led to significantly altered activity. Balancing linker length and flexibility with glycine-rich and helix-forming linker units increased 1-hexanol production activity to 350 % compared to the initial linker setup with entirely helical linkers. The study shows that strategic coupling of efficient electron supply and a selective enzyme enables previously challenging monooxygenation reactions of midchain aliphatics.

A severe head louse (Pediculus humanus capitis) infestation: Timeline, pathogen detection, kdr-gene screening and morphological anomalies of lice.

Lice are assuming an increasing importance in forensic investigations, given their capacity to provide information about an individual's care. Head louse pediculosis is a frequent condition in school-age children and can be properly controlled using topical treatments combined with good personal hygiene. Prolonged and chronic infestations may result in more serious outcomes including severe iron deficiency anaemia. We conducted entomological and laboratory investigations of a head louse infestation in a 12-year-old girl who experienced severe anaemia and subsequent death. Numerous lice were found postmortem on the head, face and neck of the patient, as well as on bedding and clothing. Analysis of nits on individual hairs determined that the louse infestation had been present for at least 166 days. The lice had some morphological traits characteristic of body lice: the third antennal segment in some specimens was distinctly longer than wide, and the apices of some paratergal plates did not extend into intersegmental membranes, while other morphological features were characteristic of head or body lice. All lice were heterozygous for the T917I kdr genotype, a marker of permethrin resistance. Nineteen (79.2%, 95%CI 59.5%-90.8%) louse DNA samples tested TaqMan positive for Acinetobacter (Moraxellales; Moraxellaceae) sp. Available information and laboratory findings are further discussed regarding their possible contribution to the negative outcome of this case. We stress the impact head louse pediculosis can have on children with limited parental attention, and how severe head louse infestation may serve as warning sign of neglect, and other high-risk situations.

Identification and characterisation of colistin-resistant Acinetobacter colistiniresistens co-producing IMP-1 and OXA-58 carbapenemases

BackgroundCarbapenem-resistant Acinetobacter is of increasing global concern because infections are challenging to treat with standard antibiotics. Here, we identified a previously uncharacterised Acinetobacter sp. clinical isolate as Acinetobacter colistiniresistens co-producing IMP-1 and OXA-58. We also examined expression of genes related to antibiotic susceptibility and drug resistance, including blaIMP. MethodsThe isolate was deposited at the National Institute of Technology and Evaluation (NITE) as Acinetobacter sp. NBRC 110496. Susceptibility was defined according to the Clinical and Laboratory Standards Institute (CLSI) breakpoints. Genomic and clonal analyses were performed to identify species and resistance genes. ResultsThe isolate was resistant to β-lactams, including broad-spectrum cephalosporins and carbapenems, polymyxins, and trimethoprim/sulfamethoxazole. Genomic analysis identified the isolate as A. colistiniresistens harbouring blaIMP-1, blaOXA-58, blaOXA-670, aac(6′)-Ib, aac(6′)-Ij, ant(3”)-II, aph(3’)-VI, msrE, mphE, and sul1. Colistin resistance was associated with the eptA-like gene, which encodes a lipid A-modifying enzyme. SNP-based phylogenetic analysis revealed that the strain clustered with other strains isolated in Japan. The IMP-1/OXA-58-producing strain described in this study has a novel integron structure surrounding blaIMP-1, aacA and sul1. ConclusionsColistin-resistant IMP-1/OXA-58-co-producing A. colistiniresistens was identified in a patient. This isolate could serve as a reservoir for carbapenemase-producing organisms. This study suggests that screening for colistin-resistant isolates is crucial to preserve colistin as a therapeutic agent for multidrug-resistant bacteria. Identification of this MDR isolate in Asia, and the danger of it spreading worldwide, should raise serious concerns.

Size-dependent ability of AtaA to immobilize cells in Acinetobacter sp. Tol 5

Microbial cells serve as efficient and environmentally friendly biocatalysts, but their stability and reusability in practical applications must often be improved through immobilization. Acinetobacter sp. Tol 5 shows high adhesiveness to materials due to its large cell surface protein AtaA, which consists of 3630 amino acids (aa). Previously, we developed a method for immobilizing bacteria using AtaA. Herein, we investigated the cell immobilization ability of in-frame deletion (IFD) mutants of AtaA with different sizes in Tol 5. Mini-AtaA, which consists of 775 aa and is functional in Escherichia coli, was produced and present on the cell surface; however, mini-AtaA showed no immobilization ability in Tol 5. A cell immobilization assay was performed with cells expressing 16 IFD mutants of AtaA with different sizes, revealing that a length of at least 1417 aa was required for the sufficient immobilization of Tol 5 cells; thus, the minimum length needed to achieve the adhesive function of AtaA varies among bacterial species. The constructed mutant library of AtaA ranging from 3630 to 775 aa will allow researchers to quickly and easily explore the optimal size of AtaA, even for bacteria newly introduced to AtaA.

Pseudomonas and Acinetobacter spp. capable of metabolizing aromatics displays multifarious plant growth promoting traits: Insights on strategizing consortium-based application to agro-ecosystems

The persistence and slow rate of natural attenuation of genotoxic and endocrine-disrupting aromatic pollutants has impacted the soil heath of agro-ecologies leading to various health and environmental issues. Ecotoxic-xenobiotic compounds degradation property along with plant growth promoting traits of microbes are beneficial for devising strategies for improved agricultural soil health and its clean-up. Present study assessed the multipartite plant growth promoting-, biocontrol- and niche adaptive- traits of five aromatic degrading soil bacteria viz. Pseudomonas bharatica CSV86T, Pseudomonas sp. C5pp, Pseudomonas sp. PP4, Pseudomonas sp. PPD and Acinetobacter sp. ISP4. Solubilization of rock phosphate (P) and feldspar (K), production of plant growth hormone (auxin; IAA), ammonia, siderophores and ACC deaminase by strain CSV86T, C5pp and PP4 were observed. Similarly, high fusaric acid resistance (MIC: 1–1.2 mg mL−1) and tolerance to salinity (5–7.5 %) was observed. In vitro application of CSV86T, C5pp, PP4 and ISP4 as consortium resulted in significant increase (P < 0.05) in the seedling shoot length (15–37 %), root length (65–150 %) and biomass (30–40 %) of wheat, mung bean and fenugreek. In-soil microcosm assay confirmed the plant growth promoting and phytoprotective ability of this consortium against toxicity of aromatics in contaminated soil. Antifungal activity against phytopathogenic fungi Magnaporthe oryzae and Aspergillus spp., ability to produce lytic enzymes and HCN suggests their biocontrol potential. Four aromatic degrading bacteria (as a consortium) exhibiting plant growth promotion and biocontrol activity could be used as an eco-friendly bioremediator-biofertilizer-biocontrol agent (bioformulation) for restoration of agricultural soil with improved crop productivity.

Design, Synthesis and Antibacterial Assessment of Active (4-Arylazo-3-Methyl-2-Thienyl) 4-Antipyrine Ketones.

The chemicals formed from antipyrines are flexible organic building blocks that are employed in the development of pharmaceuticals. By diazotizing (4-arylazo-3-hydroxy-2-thienyl) 4-antipyrine ketones 1a, 1b and 1c and (4-arylazo-3-methyl-2-thienyl) 4-antipyrine ketones (2a, 2b and 2c) further replaced with six other coupling components, a broad spectrum of hybrid molecules have been created. Mass spectra, NMR, FTIR, and elemental analyses have all been used to confirm the structures of the synthesised compounds. The antimicrobial screening was investigated by agar well diffusion and diluting the broth technique against both Gram-negative and positive-tested bacterial strains. (3-methyl-5-(phenylamino)-4-(4-tolylazo)-2-thienyl) 4-antipyrine ketone (2a) was found to be superior to Ciprofloxacin against test strains: Acinetobacter sp (34.33±1.15 mm), Listeria monocytogenes (29.33±1.15 mm) and Streptococcus sp. (19.33±1.15 mm). Also, good to moderate activities were expressed as minimum inhibitory concentration (MIC) and minimum bacterial concentration (MBC) which were recorded at 9±1 to 59.67±4.51 μg/mL and 16±4 to >512 μg/mL, respectively, using compounds 2a, 2b, and 2c. MBC/MIC ratio showed, that only, 2a and 2b have a bactericidal effect but other antipyrines with bacteriostatic strength. To conclude, it was suggested that the use of these novel synthesized (4-arylazo-3-methyl-2-thienyl) 4-antipyrine ketone derivatives molecules as a new chemical class of antimicrobial agents to perform new drug discovery in pharmaceutical preparations and medicinal research.

Bio-encapsulation of Microbial Biostimulant for Drought Resilience in Groundnut (Arachis hypogaea L.)

ABSTRACT Plant growth promoting rhizobacteria (PGPR) as microbial biostimulant promotes growth and productivity of crops under drought. However, their commercial utility is constrained by their instability and low viability in field trials. Immobilization or encapsulation of PGPR biostimulant aims to provide physical protection from stressful environment and ensure a high rate of rhizospheric colonization. In this work, we immobilized Acinetobacter calcoaceticus AC01 in 2% alginate by ionic gelation process, and evaluated for in vitro and in vivo studies. The rhizobacteria was characterized for osmotic stress, indole, phosphate solubilization, siderophore, gibberellic acid, salicylic acid, alginate and Exopolysaccharide (EPS). The encapsulated PGPR AC01 exhibited optimal moisture content of 3.57%, expansion rate of 80.62%, 95% embedding efficiency and diameter of 1.2–1.4 mm. The viability of AC01 in microcapsules remained 107CFU/g during 90 days of storage. The encapsulated PGPR biostimulant were also evaluated in Arachis hypogaea L. for drought mitigation. The inclusion of rhizobacterial microcapsules considerably induced maximum growth with significantly increased plant height, biomass, nodule count, relative water content, membrane stability index and osmolyte accumulation under drought conditions compared to non-encapsulated bacteria. Our results demonstrate that bio-encapsulation of Acinetobacter sp. AC01 in alginate matrix is a potential alternative for alleviating drought and a promising delivery system for agricultural applications.

An Active and Versatile Electron Transport System for Cytochrome P450 Monooxygenases from the Alkane Degrading Organism Acinetobacter sp. OC4.

Cytochrome P450 monooxygenases (CYPs) are valuable biocatalysts for the oxyfunctionalization of non-activated carbon-hydrogen bonds. Most CYPs rely on electron transport proteins as redox partners. In this study, the ferredoxin reductase (FdR) and ferredoxin (FD) for a cytochrome P450 monooxygenase from Acinetobacter sp. OC4 are investigated. Upon heterologous production of both proteins independently in Escherichia coli, spectral analysis showed their reduction capability towards reporter electron acceptors, e. g., cytochrome c. The individual proteins' specific activity towards cytochrome c reduction was 25 U mg-1. Furthermore, the possibility to enhance electron transfer by artificial fusion of the units was elucidated. FdR and FD were linked by helical linkers [EAAAK]n, flexible glycine linkers [GGGGS]n or rigid proline linkers [EPPPP]n of n=1-4 sequence repetitions. The system with a glycine linker (n=4) reached an appreciable specific activity of 19 U mg-1 towards cytochrome c. Moreover, their ability to drive different members of the CYP153A subfamily is demonstrated. By creating artificial self-sufficient P450s with FdR, FD, and a panel of four CYP153A representatives, effective hydroxylation of n-hexane in a whole-cell system was achieved. The results indicate this protein combination to constitute a functional and versatile surrogate electron transport system for this subfamily.

In vitro exploration of Acinetobacter strain (SG-5) for antioxidative potential and phytohormone biosynthesis in maize (Zea mays L.) cultivars differing in cadmium tolerance.

Cadmium (Cd) poses serious threats to plant growth and development, whereas the use of plant growth-promoting rhizobacteria (PGPR) has emerged a promising approach to diminish Cd retention in crops. A pot experiment was conducted to evaluate the effect of Cd tolerant strain Acinetobacter sp. SG-5 on growth, phytohormonal response, and Cd uptake of two maize cultivars (3062 and 31P41) under various Cd stress levels (0, 5, 12, 18, 26, and 30μM CdCl2). The results revealed that CdCl2 treatment significantly suppressed the seed germination and growth together with higher Cd retention in maize cultivars in a dose-dependent and cultivar-specific manner with pronounced negative effect in 31P41. However, SG-5 strain exerted positive impact by up-regulating seed germination traits, plant biomass, photosynthetic pigments, enzymatic and non-enzymatic antioxidants, endogenous hormone level indole-3-acetic acid (IAA), abscisic acid (ABA), and sustained optimal nutrient's levels in both cultivars but predominantly in Cd-sensitive one (31P41). Further, Cd-resistant PGPR decreased the formation of reactive oxygen species in terms of malondialdehyde (MDA) and hydrogen peroxide (H2O2) verified through 3, 3'-diaminobenzidine (DAB) and nitroblue tetrazolium (NBT) analysis in conjunction with reduced Cd uptake and translocation in maize root and shoots in comparison to controls, advocating its sufficiency for bacterial-assisted Cd bioremediation. In conclusion, both SG-5 inoculated cultivars exhibited maximum Cd tolerance but substantial Cd tolerance was acquired by Cd susceptible cultivar-31P41 than Cd-tolerant one (3062). Current work recommended SG-5 strain as a promising candidate for plant growth promotion and bacterial-assisted phytomanagement of metal-polluted agricultural soils.

Structure modeling-based characterization of ChnD, the 6-hydroxyhexanoate dehydrogenase from Acinetobacter sp. strain NCIMB 9871

α,ω-Dicarboxylic acids, ω-aminoalkanoic acids, and α,ω-diaminoalkanes are valuable building blocks for the production of biopolyesters and biopolyamides. One of the key steps in producing these chemicals is the oxidation of ω-hydroxycarboxylic acids using alcohol dehydrogenases (e.g., ChnD of Acinetobacter sp. NCIMB 9871). However, the reaction and structural features of these enzymes remain mostly undiscovered. Thereby, we have investigated characteristics of ChnD based on enzyme kinetics, substrate-docking simulations, and mutation studies. Kinetic analysis revealed a distinct preference of ChnD for medium chain ω-hydroxycarboxylic acids, with the highest catalytic efficiency of 18.0 mM−1s−1 for 12-hydroxydodecanoic acid among C6 to C12 ω-hydroxycarboxylic acids. The high catalytic efficiency was attributed to the positive interactions between the carboxyl group of the substrates and the guanidino group of two arginine residues (i.e., Arg62 and Arg266) in the substrate binding site. The ChnD_R62L variant showed the increased efficiency and affinity, particularly for fatty alcohols (i.e., C6–C10) and branched-chain fatty alcohols, such as 3-methyl-2-buten-1-ol. Overall, this study contributes to the deeper understanding of medium-chain primary aliphatic alcohol dehydrogenases and their applications for the production of industrially relevant chemicals such as α,ω-dicarboxylic acids, ω-aminoalkanoic acids, and α,ω-diaminoalkanes from renewable biomass.

Characteristics of Nitrogen Removal and Functional Gene Transcription of Heterotrophic Nitrification-Aerobic Denitrification Strain, Acinetobacter sp. JQ1004

In this study, the heterotrophic nitrification–aerobic denitrification strain JQ1004 was investigated in terms of its nitrogen removal mechanism and kinetic properties, laying the foundation for its application in the field of wastewater treatment. Nitrogen balance analysis revealed that the final metabolic product was N2, and approximately 54.61% of N was converted into cellular structure through assimilation. According to the fitting of the Compertz model, the maximum degradation rates of ammonia and nitrate were 7.93 mg/(L·h) and 4.08 mg/(L·h), respectively. A weakly alkaline environment was conducive to N removal, and the sensitivity of functional genes to acidic environments was amoA &gt; nirS &gt; narG. An appropriate increase in dissolved oxygen significantly enhanced heterotrophic nitrification activity, and notably, the denitrification-related functional gene narG exhibited greater tolerance to dissolved oxygen compared to nirS. The transcription level of amoA was significantly higher than that of narG or nirS, confirming that there might have been direct ammonia oxidation metabolic pathways (NH4+→NH2OH→N2) besides the complete nitrification and denitrification pathway. The annotation of nitrogen assimilation-related functional genes (including gltB, gltD, glnA, nasA, nirB, narK, nrtP, cynT, and gdhA genes) in the whole-genome sequencing analysis further confirmed the high assimilation nitrogen activity of the HN-AD strain.

Influence and mechanism of typical transition metal ions on the denitrification performance of heterotrophic nitrification-aerobic denitrification bacteria

To investigate the inhibitory effects of various transition metal ions on nitrogen removal and their underlying mechanisms, the single and combined effects of Cu2+ Ni2+ and Zn2+ on Heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria Acinetobacter sp. TAC-1 were studied in a batch experiment system. The results revealed that increasing concentrations of Cu2+ and Ni2+ had a detrimental effect on the removal of ammonium nitrogen (NH4+-N) and total nitrogen (TN). Specifically, Cu2+ concentration of 10 mg/L, the TN degradation rate was 55.09%, compared to 77.60% in the control group. Cu2+ exhibited a pronounced inhibitory effect. In contrast, Zn2+ showed no apparent inhibitory effect on NH4+-N removal and even enhanced TN removal at lower concentrations. However, when the mixed ion concentration of Zn2++Ni2+ exceeded 5 mg/L, the removal rates of NH4+-N and TN were significantly reduced. Moreover, transition metal ions did not significantly impact the removal rates of chemical oxygen demand (COD). The inhibition model fitting results indicated that the inhibition sequence was Cu2+ > Zn2+ > Ni2+. Transcriptome analysis demonstrated that metal ions influence TAC-1 activity by modulating the expression of pivotal genes, including zinc ABC transporter substrate binding protein (znuA), ribosomal protein (rpsM), and chromosome replication initiation protein (dnaA) and DNA replication of TAC-1 under metal ion stress, leading to disruptions in transcription, translation, and cell membrane structure. Finally, a conceptual model was proposed by us to summarize the inhibition mechanism and possible response strategies of TAC-1 bacteria under metal ion stress, and to address the lack of understanding regarding the influence mechanism of TAC-1 on nitrogen removal in wastewater co-polluted by metal and ammonia nitrogen. The results provided practical guidance for the management of transition metal and ammonia nitrogen co-polluted water bodies, as well as the removal of high nitrogen.

Transcriptome analysis of Acinetobacter calcoaceticus HX09 strain with outstanding crude-oil-degrading ability.

Microbial remediation plays a pivotal role in the elimination of petroleum pollutants, making it imperative to investigate the capabilities of microorganisms in degrading petroleum. The present study describes the isolation of a promising strain, Acinetobacter sp. HX09, from petroleum-contaminated water. GC-MS analysis revealed a remarkable removal efficiency for short and medium-chain alkanes, with a rate of approximately 64% after a 7-days incubation at 30 °C. Transcriptome analysis of HX09 exhibited a predominant upregulation in gene expression levels by the induce of crude oil. Notably, genes such as alkane 1-monooxygenase, dehydrogenases and fatty acid metabolic enzymes exhibited fold changes range from 3.16 to 1.3. Based on the alkB gene sequences in HX09, the Phyre2 algorithm generated a three-dimensional structure that exhibited similarity to segments of acyl coenzyme desaturases and acyl lipid desaturases. Furthermore, three biodegradation-related gene clusters were predicted in HX09 based on the reference genome sequence. These findings contribute to our understanding of the hydrocarbon-degrading mechanisms employed by Acinetobacter species and facilitate the development of effective remediation strategies for crude oil- polluted environments.

Osmolyte-producing microbial biostimulants regulate the growth of Arachis hypogaea L. under drought stress

Globally, drought stress poses a significant threat to crop productivity. Improving the drought tolerance of crops with microbial biostimulants is a sustainable strategy to meet a growing population’s demands. This research aimed to elucidate microbial biostimulants’ (Plant Growth Promoting Rhizobacteria) role in alleviating drought stress in oil-seed crops. In total, 15 bacterial isolates were selected for drought tolerance and screened for plant growth-promoting (PGP) attributes like phosphate solubilization and production of indole-3-acetic acid, siderophore, hydrogen cyanide, ammonia, and exopolysaccharide. This research describes two PGPR strains: Acinetobacter calcoaceticus AC06 and Bacillus amyloliquefaciens BA01. The present study demonstrated that these strains (AC06 and BA01) produced abundant osmolytes under osmotic stress, including proline (2.21 and 1.75 µg ml− 1), salicylic acid (18.59 and 14.21 µg ml− 1), trehalose (28.35 and 22.74 µg mg− 1 FW) and glycine betaine (11.35 and 7.74 mg g− 1) respectively. AC06 and BA01 strains were further evaluated for their multifunctional performance by inoculating in Arachis hypogaea L. (Groundnut) under mild and severe drought regimes (60 and 40% Field Capacity). Inoculation with microbial biostimulants displayed distinct osmotic-adjustment abilities of the groundnut, such as growth parameters, plant biomass, photosynthetic pigments, relative water content, proline, and soluble sugar in respective to control during drought. On the other hand, plant sensitivity indexes such as electrolyte leakage and malondialdehyde (MDA) contents were decreased as well as cooperatively conferred plant drought tolerance by induced alterations in stress indicators such as catalase (CAT), ascorbate peroxidase (APX), and superoxide dismutase (SOD). Thus, Acinetobacter sp. AC06 and Bacillus sp. BA01 can be considered as osmolyte producing microbial biostimulants to simultaneously induce osmotic tolerance and metabolic changes in groundnuts under drought stress.

The elimination of two restriction enzyme genes allows for electroporation-based transformation and CRISPR-Cas9-based base editing in the non-competent Gram-negative bacterium Acinetobacter sp. Tol 5.

Environmental isolates are promising candidates for new chassis of synthetic biology because of their inherent capabilities, which include efficiently converting a wide range of substrates into valuable products and resilience to environmental stresses; however, many remain genetically intractable and unamenable to established genetic tools tailored for model bacteria. Acinetobacter sp. Tol 5, an environmentally isolated Gram-negative bacterium, possesses intriguing properties for use in synthetic biology applications. Despite the previous development of genetic tools for the engineering of strain Tol 5, its genetic manipulation has been hindered by low transformation efficiency via electroporation, rendering the process laborious and time-consuming. This study demonstrated the genetic refinement of the Tol 5 strain, achieving efficient transformation via electroporation. We deleted two genes encoding type I and type III restriction enzymes. The resulting mutant strain not only exhibited marked efficiency of electrotransformation but also proved receptive to both in vitro and in vivo DNA assembly technologies, thereby facilitating the construction of recombinant DNA without reliance on intermediate Escherichia coli constructs. In addition, we successfully adapted a CRISPR-Cas9-based base-editing platform developed for other Acinetobacter species. Our findings provide genetic modification strategies that allow for the domestication of environmentally isolated bacteria, streamlining their utilization in synthetic biology applications.IMPORTANCERecent synthetic biology has sought diverse bacterial chassis from environmental sources to circumvent the limitations of laboratory Escherichia coli strains for industrial and environmental applications. One of the critical barriers in cell engineering of bacterial chassis is their inherent resistance to recombinant DNA, propagated either in vitro or within E. coli cells. Environmental bacteria have evolved defense mechanisms against foreign DNA as a response to the constant threat of phage infection. The ubiquity of phages in natural settings accounts for the genetic intractability of environmental isolates. The significance of our research is in demonstrating genetic modification strategies for the cell engineering of such genetically intractable bacteria. This research marks a pivotal step in the domestication of environmentally isolated bacteria, promising candidates for emerging synthetic biology chassis. Our work thus significantly contributes to advancing their applications across industrial, environmental, and biomedical fields.