Genomic insights and anti-phytopathogenic potential of siderophore metabolome of endolithic Nocardia mangyaensis NH1.

Actinobacteria are one of the predominant groups that successfully colonize and survive in various aquatic, terrestrial and rhizhospheric ecosystems. Among actinobacteria, Nocardia is one of the most important agricultural and industrial bacteria. Screening and isolation of Nocardia related bacteria from extreme habitats such as endolithic environments are beneficial for practical applications in agricultural and environmental biotechnology. In this work, bioinformatics analysis revealed that a novel strain Nocardia mangyaensis NH1 has the capacity to produce structurally varied bioactive compounds, which encoded by non-ribosomal peptide synthases (NRPS), polyketide synthase (PKS), and post-translationally modified peptides (RiPPs). Among NRPS, five gene clusters have a sequence homology with clusters encoding for siderophore synthesis. We also show that N. mangyaensis NH1 accumulates both catechol- and hydroxamate-type siderophores simultaneously under iron-deficient conditions. Untargeted LC-MS/MS analysis revealed a variety of metabolites, including siderophores, lipopeptides, cyclic peptides, and indole-3-acetic acid (IAA) in the culture medium of N. mangyaensis NH1 grown under iron deficiency. We demonstrate that four CAS (chrome azurol S)-positive fractions display variable affinity to metals, with a high Fe3+ chelating capability. Additionally, three of these fractions exhibit antioxidant activity. A combination of iron scavenging metabolites produced by N. mangyaensis NH1 showed antifungal activity against several plant pathogenic fungi. We have shown that the pure culture of N. mangyaensis NH1 and its metabolites have no adverse impact on Arabidopsis seedlings. The ability of N. mangyaensis NH1 to produce siderophores with antifungal, metal-chelating, and antioxidant properties, when supplemented with phytohormones, has the potential to improve the release of macro- and micronutrients, increase soil fertility, promote plant growth and development, and enable the production of biofertilizers across diverse soil systems.

The actinomycete Kitasatospora sp. SeTe27, subjected to adaptive laboratory evolution (ALE) in the presence of selenite, varies its cellular morphology, redox stability, and tolerance to the toxic oxyanion.

The effects of oxyanions selenite (SeO32-) in soils are of high concern in ecotoxicology and microbiology as they can react with mineral particles and microorganisms. This study investigated the evolution of the actinomycete Kitasatospora sp. SeTe27 in response to selenite. To this aim, we used the Adaptive Laboratory Evolution (ALE) technique, an experimental approach that mimics natural evolution and enhances microbial fitness for specific growth conditions. The original strain (wild type; WT) isolated from uncontaminated soil gave us a unique model system as it has never encountered the oxidative damage generated by the prooxidant nature of selenite. The WT strain exhibited a good basal level of selenite tolerance, although its growth and oxyanion removal capacity were limited compared to other environmental isolates. Based on these premises, the WT and the ALE strains, the latter isolated at the end of the laboratory evolution procedure, were compared. While both bacterial strains had similar fatty acid profiles, only WT cells exhibited hyphae aggregation and extensively produced membrane-like vesicles when grown in the presence of selenite (challenged conditions). Conversely, ALE selenite-grown cells showed morphological adaptation responses similar to the WT strain under unchallenged conditions, demonstrating the ALE strain improved resilience against selenite toxicity. Whole-genome sequencing revealed specific missense mutations in genes associated with anion transport and primary and secondary metabolisms in the ALE variant. These results were interpreted to show that some energy-demanding processes are attenuated in the ALE strain, prioritizing selenite bioprocessing to guarantee cell survival in the presence of selenite. The present study indicates some crucial points for adapting Kitasatospora sp. SeTe27 to selenite oxidative stress to best deal with selenium pollution. Moreover, the importance of exploring non-conventional bacterial genera, like Kitasatospora, for biotechnological applications is emphasized.

Malignance-restriction activity exhibited by bioactive compounds of selected actinobacteria as silver nanoparticles against A549 lung cancer cell lines.

This article deals with the antibacterial and anticancer potential of secondary metabolites produced by actinomycetes also reported as actinobacteria, Microbacterium proteolyticum (MN560041), and Streptomycetes rochei, where preliminary studies were done with the well diffusion method. These actinobacteria's silver nanoparticles were synthesized and characterized using transmission electron microscopy (TEM) and UV-Visible spectroscopy. Anticancer was measured using the MTT test, reactive oxygen species (ROS) generation measured with DCFDA, mitochondrial membrane potential (MMP) measurement, and DAPI fluorescence intensity activity was measured in treated and non-treated cancerous cells. The IC50 value for 5-FU (a), LA2(O) (b), LA2(R) (c), LA2(ON) (d), and LA2(RN) (e) was obtained at 3.91 µg/mL (52.73% cell viability), 56.12 µg/mL (52.35% cell viability), 44.90 µg/mL (52.3% cell viability), 3.45 µg/mL (50.25% cell viability), and 8.05 µg/mL (48.72% cell viability), respectively. TEM micrographs revealed discrete, well-separated AgNPs particles of size 7.88 ± 2 to 12.86 ± 0.24 nm. Gas chromatography-mass spectrometry was also performed to detect the compounds in bioactive metabolites where n-hexadecanoic acid was obtained as the most significant one. MTT test showed a substantial decline in A549 cell viability (up to 48.72%), 2.75-fold increase in ROS generation was noticed in comparison to untreated A549 lung cancer cells when measured with DCFDA. A total of 0.31-fold decrease in MMP and 1.74-fold increase in DAPI fluorescence intensity compared to untreated A549 lung cancer cells suggests that the synthesized nanoparticles promote apoptosis in cancerous cells. Our findings suggests that the secondary metabolites of M. proteolyticum and S. rochei in nanoparticle form can be used as a significant compound against lung cancers.

Screening biosurfactant-producing actinomycetes: Identification of Streptomyces sp. RP1 as a potent species for bioremediation.

This study aimed to isolate biosurfactant-producing and hydrocarbon-degrading actinomycetes from different soils using glycerol-asparagine and starch-casein media with an antifungal agent. The glycerol-asparagine agar exhibited the highest number of actinomycetes, with a white, low-opacity medium supporting pigment production and high growth. Biosurfactant analyses, such as drop collapse, oil displacement, emulsification, tributyrin agar test, and surface tension measurement, were conducted. Out of 25 positive isolates, seven could utilize both olive oil and black oil for biosurfactant production, and only isolate RP1 could produce biosurfactant when grown in constrained conditions with black oil as the sole carbon source and inducer, demonstrating in situ bioremediation potential. Isolate RP1 from oil-spilled garden soil is Gram-staining-positive with a distinct earthy odor, melanin formation, and white filamentous colonies. It has a molecular size of ~621 bp and 100% sequence similarity to many Streptomyces spp. Morphological, biochemical, and 16 S rRNA analysis confirmed it as Streptomyces sp. RP1, showing positive results in all screenings, including high emulsification activity against kerosene (27.2%) and engine oil (95.8%), oil displacement efficiency against crude oil (7.45 cm), and a significant reduction in surface tension (56.7 dynes/cm). Streptomyces sp. RP1 can utilize citrate as a carbon source, tolerate sodium chloride, resist lysozyme, degrade petroleum hydrocarbons, and produce biosurfactant at 37°C in a 15 mL medium culture, indicating great potential for bioremediation and various downstream industrial applications with optimization.

Antagonistic Potentiality of Actinomycete-Derived Extract with Anti-Biofilm, Antioxidant, and Cytotoxic Capabilities as a Natural Combating Strategy for Multidrug-Resistant ESKAPE Pathogens.

The global increase in multidrug-resistant (MDR) bacteria has inspired researchers to develop new strategies to overcome this problem. In this study, 23 morphologically different, soil-isolated actinomycete cultures were screened for their antibacterial ability against MDR isolates of ESKAPE pathogens. Among them, isolate BOGE18 exhibited a broad antibacterial spectrum, so it was selected and identified based on cultural, morphological, physiological, and biochemical characteristics. Chemotaxonomic analysis was also performed together with nucleotide sequencing of the 16S rRNA gene, which showed this strain to have identity with Streptomyces lienomycini. The ethyl acetate extract of the cell-free filtrate (CFF) of strain BOGE18 was evaluated for its antibacterial spectrum, and the minimum inhibitory concentration (MIC) ranged from 62.5 to 250 µg/ml. The recorded results from the in vitro anti-biofilm microtiter assay and confocal laser scanning microscopy (CLSM) of sub-MIC concentrations revealed a significant reduction in biofilm formation in a concentration-dependent manner. The extract also displayed significant scavenging activity, reaching 91.61 ± 4.1% and 85.06 ± 3.14% of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis( 3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), respectively. A promising cytotoxic ability against breast (MCF-7) and hepatocellular (HePG2) cancer cell lines was obtained from the extract with IC50 values of 47.15 ± 13.10 and 122.69 ± 9.12 µg/ml, respectively. Moreover, based on gas chromatography-mass spectrometry (GC-MS) analysis, nine known compounds were detected in the BOGE18 extract, suggesting their contribution to the multitude of biological activities recorded in this study. Overall, Streptomyces lienomycini BOGE18-derived extract is a good candidate for use in a natural combating strategy to prevent bacterial infection, especially by MDR pathogens.

Streptomyces rochei MS-37 as a Novel Marine Actinobacterium for Green Biosynthesis of Silver Nanoparticles and Their Biomedical Applications.

Periodontitis, as one of the most common diseases on a global scale, is a public health concern. Microbial resistance to currently available antimicrobial agents is becoming a growing issue in periodontal treatment. As a result, it is critical to develop effective and environmentally friendly biomedical approaches to overcome such challenges. The investigation of Streptomyces rochei MS-37's performance may be the first of its kind as a novel marine actinobacterium for the green biosynthesis of silver nanoparticles (SNPs) and potentials as antibacterial, anti-inflammatory, antibiofilm, and antioxidant candidates suppressing membrane-associated dental infections. Streptomyces rochei MS-37, a new marine actinobacterial strain, was used in this study for the biosynthesis of silver nanoparticles for various biomedical applications. Surface plasmon resonance spectroscopy showed a peak at 429 nm for the SNPs. The SNPs were spherical, tiny (average 23.2 nm by TEM, 59.4 nm by DLS), very stable (-26 mV), and contained capping agents. The minimum inhibitory concentrations of the SNPs that showed potential antibacterial action ranged from 8 to 128 µg/mL. Periodontal pathogens were used to perform qualitative evaluations of microbial adhesion and bacterial penetration through guided tissue regeneration membranes. The findings suggested that the presence of the SNPs could aid in the suppression of membrane-associated infection. Furthermore, when the anti-inflammatory action of the SNPs was tested using nitric oxide radical scavenging capacity and protein denaturation inhibition, it was discovered that the SNPs were extremely efficient at scavenging nitric oxide free radicals and had a strong anti-denaturation impact. The SNPs were found to be more cytotoxic to CAL27 than to human peripheral blood mononuclear cells (PBMCs), with IC50 values of 81.16 µg/mL in PBMCs and 34.03 µg/mL in CAL27. This study's findings open a new avenue for using marine actinobacteria for silver nanoparticle biosynthesis, which holds great promise for a variety of biomedical applications, in particular periodontal treatment.

Actinobacteria bioaugmentation and substrate evaluation for biobeds useful for the treatment of atrazine residues in agricultural fields.

Biopurification systems (BPS) or biobeds are bioprophylaxis systems to prevent pesticide point-source contamination, whose efficiency relies mostly on the pesticide removal capacity of the biomixture, the majority component of a BPS. The adaptation of the components of the biomixtures to local availabilities is a key aspect to ensure the sustainability of the system. In this work, the removal of atrazine (ATZ) was evaluated in biomixtures formulated with three sugarcane by-products as alternative lignocellulosic substrates. Based on the capacity of actinobacteria to tolerate and degrade diverse pesticides, the effect of biomixtures bioaugmentation with actinobacteria was evaluated as a strategy to enhance the depuration capacity of biobeds. Also, the effect of ATZ and/or the bioaugmentation on microbial developments and enzymatic activities were studied. The biomixtures formulated with bagasse, filter cake, or harvest residue, reached pesticide removal values of 37-41% at 28 d of incubation, with t1/2 between 37.9 ± 0.4 d and 52.3 ± 0.4 d. The bioaugmentation with Streptomyces sp. M7 accelerated the dissipation of the pesticide in the biomixtures, reducing ATZ t1/2 3-fold regarding the controls, and achieving up to 72% of ATZ removal. Atrazine did not exert a clear effect on microbial developments, although most of the microbial counts were less in the contaminated biomixtures at the end of the assay. The bioaugmentation improved the development of the microbiota in general, specially actinobacteria and fungi, regarding the non-bioaugmented systems. The inoculation with Streptomyces sp. M7 enhanced acid phosphatase activity and/or reversed a possible effect of the pesticide over this enzymatic activity.

Iron-Induced Respiration Promotes Antibiotic Resistance in Actinomycete Bacteria.

The bacterial response to antibiotics eliciting resistance is one of the key challenges in global health. Despite many attempts to understand intrinsic antibiotic resistance, many of the underlying mechanisms still remain elusive. In this study, we found that iron supplementation promoted antibiotic resistance in Streptomyces coelicolor. Iron-promoted resistance occurred specifically against bactericidal antibiotics, irrespective of the primary target of antibiotics. Transcriptome profiling revealed that some genes in the central metabolism and respiration were upregulated under iron-replete conditions. Iron supported the growth of S. coelicolor even under anaerobic conditions. In the presence of potassium cyanide, which reduces aerobic respiration of cells, iron still promoted respiration and antibiotic resistance. This suggests the involvement of a KCN-insensitive type of respiration in the iron effect. This phenomenon was also observed in another actinobacterium, Mycobacterium smegmatis. Taken together, these findings provide insight into a bacterial resistance strategy that mitigates the activity of bactericidal antibiotics whose efficacy accompanies oxidative damage by switching the respiration mode. IMPORTANCE A widely investigated mode of antibiotic resistance occurs via mutations and/or by horizontal acquisition of resistance genes. In addition to this acquired resistance, most bacteria exhibit intrinsic resistance as an inducible and adaptive response to different classes of antibiotics. Increasing attention has been paid recently to intrinsic resistance mechanisms because this may provide novel therapeutic targets that help rejuvenate the efficacy of the current antibiotic regimen. In this study, we demonstrate that iron promotes the intrinsic resistance of aerobic actinomycetes Streptomyces coelicolor and Mycobacterium smegmatis against bactericidal antibiotics. A surprising role of iron to increase respiration, especially in a mode of using less oxygen, appears a fitting strategy to cope with bactericidal antibiotics known to kill bacteria through oxidative damage. This provides new insights into developing antimicrobial treatments based on the availability of iron and oxygen.

Biodegradation of used motor oil by Streptomyces ginkgonis KM-1-2, isolated from soil polluted by waste oils in the region of Azzaba (Skikda-Algeria).

Actinobacteria have many properties that make them good candidates for the bioremediation of sites contaminated by several organic and inorganic pollutants. However, studies on the biodegradation of used motor oils by Actinobacteria, compared to other bacteria, remain little studied. Actinobacteria were isolated from soil contaminated with used motor oils and sewage sludge in order to select a species that can effectively degrade such pollutants. This study aims to assess their degradation capacity of the various hydrocarbon fractions contained in the oil by gas chromatography-mass spectrometry (GC-MS). Five Actinobacteria isolates were isolated by the enrichment method. The S1.1. A strain was considered the best biosurfactant producing strain. It presents the highest emulsification index compared to other isolated Actinobacteria (82.6%). Phenotypic and molecular identification by sequencing the 16 S rDNA gene makes it possible to assign this isolate to the species Streptomyces ginkgonis KM-1-2. Gravimetric analysis results of biodegraded used motor oil indicated that this strain is capable of degrading 76.4% of an initial 50 ml/l concentration of used motor oil after 4 weeks of incubation. The results of GC-MS analysis of the residual motor oil showed that strain S1.1. A degraded some long and intermediate chain alkanes completely or to shorter fractions. The Streptomyces ginkgonis strain KM-1-2 was also able to degrade certain alkylated mono-aromatic hydrocarbons linked to benzene, as well as certain alkylated and non-alkylated polycyclic aromatic hydrocarbons linked to anthracene, naphthalene, phenanthrene, fluorene, and azulene. This strain exhibited the highest emulsification index at 82.6%. This bacterium shows a significant biodegradation capacity and could, consequently, be used in the processes of bioaugmentation of sites contaminated by these oils.

Screening of Natural Products Inhibitors of SARS-CoV-2 Entry.

The COVID-19 pandemic has led to the search for new molecules with antiviral activity against SARS-CoV-2. The entry of the virus into the cell is one of the main targets for inhibiting SARS-CoV-2 infection. Natural products are an important source of new therapeutic alternatives against diseases. Pseudotyped viruses allow the study of SARS-CoV-2 viral entry inhibitors, and due to their simplicity, they allow the screening of a large number of antiviral candidates in Biosafety Level 2 facilities. We used pseudotyped HIV-1 with the D614G SARS-CoV-2 spike glycoprotein to test its ability to infect ACE2-expressing HEK 293T cells in the presence of diverse natural products, including 21 plant extracts, 7 essential oils, and 13 compounds from plants and fungi. The 50% cytotoxic concentration (CC50) was evaluated using the resazurin method. From these analyses, we determined the inhibitory activity of the extract of Stachytarpheta cayennensis, which had a half-maximal inhibitory concentration (IC50) of 91.65 µg/mL, a CC50 of 693.5 µg/mL, and a selectivity index (SI) of 7.57, indicating its potential use as an inhibitor of SARS-CoV-2 entry. Moreover, our work indicates the usefulness of the pseudotyped-virus system in the screening of SARS-CoV-2 entry inhibitors.

Correlation of bacterial community with phosphorus fraction drives discovery of Actinobacteria involved soil phosphorus transformation during the trichlorfon degradation.

Trichlorfon (TCF) is a broad-spectrum phosphorus (P)-containing pesticide, yet its effects on soil P fraction transformation and bacterial communities during the TCF degradation in soils is unknown. In this study, we investigated soil TCF degradation behavior at different contents of 50, 100 and 200 mg/kg, and analyzed residual TCF contents and metabolites by gas chromatography mass spectrometry after 216-h incubation. Our results suggested that TCF was gradually degraded in soils and was be initially hydrolyzed to dichlorvos via P-C bond cleavage and then other P-containing metabolites. By analyzing different P fractions and soil microbial community composition, we found significant increases of soil available phosphorus contents from 2.76 mg/kg (control) to 3.23 mg/kg (TCF-50), 5.12 mg/kg (TCF-100) and 5.72 mg/kg (TCF-200), respectively. Inorganic CaCl2-P was easily and instantly transformed to primary mineral inorganic P (Pi) forms of HCl-P and citrate-P, while the proportion of enzyme-P (a labile organic P) fluctuated throughout TCF degradation process. Soil available P contents and Pi fractions were significantly correlated with the relative abundance of Actinobacteria. These results highlighted that Actinobacteria is the dominant soil species utilizing TCF as P sources to increase its community richness, and subsequently affect the transformation of P fractions to regulate soil P cycle. Our study gives new understanding on the microorganisms can involve soil P transformation during organophosphorus pesticides degradation in soils, highlighting the importance of bacteria in P transformation and pesticides soil decontamination.

Rat alpha glucosidase inhibitor and phytochemicals activities of endophytic actinobacteria from Ficus deltoidea.

Ficus deltoidea is a medicinal plant that has high endophytic actinobacteria diversity. Endophytic actinobacteria play an important role in producing various types of bioactive compounds including α-glucosidase inhibitor. Screening of 40 endophytic actinobacteria isolates from F. deltoidea showed that 77% of them had inhibitory activity against rat α-glucosidase. The 64% of isolates that have rat α-glucosidase inhibitor activity were derived from leaves. TBL 7, TBL 24, TBS 3, TBS 17 and TBR 20 have high activity. Based on the molecular identification of the 16S rRNA gene, five selected isolates have similarity with Streptomyces spp. The aqueous and n-hexane extracts of TBL 7 isolates had the lowest IC50 values of 159.25 µg/ml and 118.52 µg/ml, respectively. Qualitative phytochemical analysis showed that aqueous and n-hexane extracts of TBL 7 contained flavonoids, phenols, alkaloids, triterpenoids, tannins, and saponins. These results showed that endophytic actinobacteria from F. detoidea have the potential to be developed as α-glucosidase inhibitor.

Effect of anaerobic phases length on denitrifying dephosphatation biocenosis - a case study of IFAS-MBSBBR.

BACKGROUND: The study aimed to evaluate the influence of the duration times of anaerobic phases on the bacterial biocenosis characterisation while denitrifying dephosphatation in the Integrated Fixed-Film Activated Sludge - Moving-Bed Sequencing Batch Biofilm Reactor (IFAS-MBSBBR). The experiment was conducted in a laboratory model. The study consisted of four series, which differed in terms of the ratio of the anaerobic phases. duration concerning the overall reaction time in the cycle. The anaerobic phases covered from 18 to 30% of the whole cycle duration. During the reactor performance that took 9 months, the influent and effluent were monitored by analysis of COD, TKN, NH4-N, NO2-N, NO3-N, TP, PO4-P, pH, alkalinity and the phosphorus uptake batch tests. Characterisation of the activated sludge and the biofilm biocenosis was based on fluorescent in situ hybridisation (identification of PAO and GAO) and the denaturing gradient gel electrophoresis patterns. RESULTS: The organic compounds removal was high (more than 95.7%) independently of cycle configuration. The best efficiency for nitrogen (91.1%) and phosphorus (98.8%) removal was achieved for the 30% share of the anaerobic phases in the reaction time. Denitrifying PAO (DPAO) covered more than 90% of PAO in the biofilm and usually around 70% of PAO in the activated sludge. A substantial part of the polyphosphate accumulating organisms (PAO) community were Actinobacteria. The denitrifying dephosphatation activity was performed mainly by Accumulibacter phosphatis. CONCLUSIONS: High nutrient removal efficiencies may be obtained in IFAS-MBSBBR using the denitrifying dephosphatation process. It was found that the length of anaerobic phases influenced denitrification and the biological phosphorus removal. The extension of the anaerobic phases duration time in the reaction time caused an increase in the percentage share of denitrifying PAO (DPAO) in PAO. The biocenosis of the biofilm and the activated sludge reveal different species patterns and domination of the EBPR community.

Innovative, ecofriendly biosorbent-biodegrading biofilms for bioremediation of oil- contaminated water.

Immobilization of microorganisms capable of degrading specific contaminants significantly promotes bioremediation processes. In this study, innovative and ecofriendly biosorbent-biodegrading biofilms have been developed in order to remediate oil-contaminated water. This was achieved by immobilizing hydrocarbon-degrading gammaproteobacteria and actinobacteria on biodegradable oil-adsorbing carriers, based on polylactic acid and polycaprolactone electrospun membranes. High capacities for adhesion and proliferation of bacterial cells were observed by scanning electron microscopy. The bioremediation efficiency of the systems, tested on crude oil and quantified by gas chromatography, showed that immobilization increased hydrocarbon biodegradation by up to 23 % compared with free living bacteria. The resulting biosorbent biodegrading biofilms simultaneously adsorbed 100 % of spilled oil and biodegraded more than 66 % over 10 days, with limited environmental dispersion of cells. Biofilm-mediated bioremediation, using eco-friendly supports, is a low-cost, low-impact, versatile tool for bioremediation of aquatic systems.

Effects of dietary grape pomace on the intestinal microbiota and growth performance of weaned piglets.

Grape pomace (GP) is an abundant by-product from wine production and is rich in phenolic compounds, unsaturated fatty acids, dietary fibre and beneficial bacteria. In this study, weaned piglets were fed a basic diet supplemented with 5% GP for 4 weeks. Compared with those in the control (CON) group, it was found that the proportion of Lactobacillus delbrueckii, Olsenella umbonata and Selenomonas bovis in the caecum and the villus height and villus height/crypt depth ratio (VCR) of the jejunum were both significantly increased in the GP group (p < 0.05). Meanwhile, at the mRNA expression level, several proinflammatory cytokines (IL-1ß, IL-8, IL-6 and TNF-α) were significantly downregulated (p < 0.05) in piglet caecal tissue, and the short-chain fatty acid receptors (GPR41 and GPR43) were not significantly upregulated. In contrast, the levels of IgG was significantly increased (p < 0.05) in the sera of weaned piglets in the GP group. However, no difference in growth performance between the two groups of piglets was detected. These results show that GP had no adverse effects on the growth performance of piglets, but GP can promote the content of some beneficial bacteria in the caecum; this effect is conducive to improving the disease resistance potential of piglets.

Exploring the fermentation characteristics of a newly isolated marine bacteria strain, Gordonia terrae TWRH01 for carotenoids production.

Carotenoids serve as one of the most important group of naturally-occurring lipid-soluble pigments which exhibit great biological activities such as antioxidant, anti-inflammatory and provitamin A activities. Owing to their advantageous health effects, carotenoids are widely applied in various industries. Microbial carotenoids synthesis therefore has attracted increasing attention in recent years. In the present study, a marine microorganism originally isolated from seawater in northern Taiwan was determined to be a strain of Gordonia terrae based on its 16S rRNA gene sequence. The strain G. terrae TWRH01 has the ability to synthesize and accumulate the intracellular pigments was identified by gas chromatography-mass spectrometry (GC-MS). The biochemical production characteristics of this strain were studied by employing different fermentation strategies. Findings suggested that G. terrae TWRH01 can actively grow and efficiently synthesize carotenoids in medium adjusted to pH 7 containing 16 g L-1 sucrose as the carbon source, 16 g L-1 yeast extract as the nitrogen source, 0.6 M NaCl concentration, and supplemented with 0.45% (v/v) 1 M CaCl2. Results revealed that the optimization of fermentation yielded 15.29 g L-1 dry biomass and 10.58 µmol L-1 relative ß-carotene concentration. According to GC-MS analysis, the orange-red colored pigments produced were identified as carotenoid derivatives, mainly echinenone and adonixanthin 3'-ß-d-glucoside. Therefore, the new bacterial strain showed a highly potential bioresource for the commercial production of natural carotenoids.

Screening of new cell cycle suppressive compounds from marine-derived microorganisms in Chinese hamster ovary cells.

Monoclonal antibodies (mAbs) are active pharmaceutical ingredients in antibody drugs, produced mainly using recombinant Chinese hamster ovary (CHO) cells. The regulation of recombinant CHO cell proliferation can improve the productivity of heterologous proteins. Chemical compound approaches for cell cycle regulation have the advantages of simplicity and ease of use in industrial processes. However, CHO cells have genetic and phenotypic diversity, and the effects of such compounds might depend on cell line and culture conditions. Increasing the variety of cell cycle inhibitors is a promising strategy to overcome the dependency. Marine microorganisms are a vast and largely undeveloped source of secondary metabolites with physiological activity. In this study, we focused on secondary metabolites of marine microorganisms and evaluated their effectiveness as cell cycle inhibitory compounds. Of 720 extracts from microorganisms (400 actinomycetes and 320 filamentous fungi) collected from the Okinawan Sea, we identified nine extracts that decreased the specific growth rate and increased the specific production rate without reducing cell viability. After fractionating the extracts, the components of active fractions were estimated using time-of-flight mass spectrometry analysis. Then, four compounds, including staurosporine and undecylprodigiosin were deduced to be active compounds. These compounds have been reported to exert a cell cycle inhibitory effect on mammalian cells. These compounds might serve as additives to improve mAb production in CHO cells. This study indicates that secondary metabolites of marine microorganisms are a useful source for new cell cycle inhibitory compounds that can increase mAb production in CHO cells.

Bioremediation of soils saturated with spilled crude oil.

A desert soil sample was saturated with crude oil (17.3%, w/w) and aliquots were diluted to different extents with either pristine desert or garden soils. Heaps of all samples were exposed to outdoor conditions through six months, and were repeatedly irrigated with water and mixed thoroughly. Quantitative determination of the residual oil in the samples revealed that oil-bioremediation in the undiluted heaps was nearly as equally effective as in the diluted ones. One month after starting the experiment. 53 to 63% of oil was removed. During the subsequent five months, 14 to 24% of the oil continued to be consumed. The dynamics of the hydrocarbonoclastic bacterial communities in the heaps was monitored. The highest numbers of those organisms coordinated chronologically with the maximum oil-removal. Out of the identified bacterial species, those affiliated with the genera Nocardioides (especially N. deserti), Dietzia (especially D. papillomatosis), Microbacterium, Micrococcus, Arthrobacter, Pseudomonas, Cellulomonas, Gordonia and others were main contributors to the oil-consumption. Some species, e.g. D. papillomatosis were minor community constituents at time zero but they prevailed at later phases. Most isolates tolerated up to 20% oil, and D. papillomatosis showed the maximum tolerance compared with all the other studied isolates. It was concluded that even in oil-saturated soil, self-cleaning proceeds at a normal rate. When pristine soil receives spilled oil, indigenous microorganisms suitable for dealing with the prevailing oil-concentrations become enriched and involved in oil-biodegradation.

Influence of Leifsonia sp. on U(VI) removal efficiency and the Fe-U precipitates by zero-valent iron.

Zero-valent iron (ZVI) has been widely applied to the remediation of uranium (U)-contaminated water. Notably, indigenous bacteria may possess potential positive or unfavorable influence on the mechanism and stability of Fe-U precipitates. However, the focus of the researches in this field has mainly been on physical and/or chemical aspects. In this study, batch experiments were conducted to explore the effects of an indigenous bacterium (Leifsonia sp.) on Fe-U precipitates and the corresponding removal efficiency by ZVI under different environmental factors. The results showed that the removal rate and capacity of U(VI) was significantly inhibited and decreased by ZVI when the pH increased to near-neutral level (pH = 6~8). However, in the ZVI + Leifsonia sp. coexistence system, the U(VI) removal efficiency were maintained at high levels (over 90%) within the experimental scope (pH = 3~8). This revealed that Leifsonia sp. had a synergistic effect on U(VI) remove by ZVI. According to scanning electron microscope and energy dispersive X-ray detector (SEM-EDX) analysis, dense scaly uranium-phosphate precipitation was observed on ZVI + Leifsonia sp. surface. The X-photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR) analysis indicated that Leifsonia sp. facilitated the generation of U(VI)-phosphates precipitates. The X-ray diffraction (XRD) analyses further revealed that new substances, such as (Fe(II)Fe(III)2(PO4)2(OH)2), Fe(II)(UO2)2(PO4)2·8H2O, Fe(II)Fe(III)5(PO4)4(OH)2·4H2O, etc., were produced in the coexisting system of ZVI and Leifsonia sp. This study provides new insights on the feasibility and validity of site application of ZVI to U(VI)-contaminated subsurface water in situ. Graphical abstract.

Low-Abundance Dietzia Inhabiting a Water-Flooding Oil Reservoir and the Application Potential for Oil Recovery.

With the development of molecular ecology, increasing low-abundance microbial populations were detected in oil reservoirs. However, our knowledge about the oil recovery potential of these populations is lacking. In this study, the oil recovery potential of low-abundance Dietzia that accounts for less than 0.5% in microbial communities of a water-flooding oil reservoir was investigated. On the one hand, Dietzia sp. strain ZQ-4 was isolated from the water-flooding reservoir, and the oil recovery potential was evaluated from the perspective of metabolisms and oil-displacing test. On the other hand, the strain has alkane hydroxylase genes alkB and P450 CYP153 and can degrade hydrocarbons and produce surfactants. The core-flooding test indicated that displacing fluid with 2% ZQ-4 fermentation broth increased 18.82% oil displacement efficiency, and in situ fermentation of ZQ-4 increased 1.97% oil displacement efficiency. Furthermore, the responses of Dietzia in the reservoir accompanied by the nutrient stimulation process was investigated and showed that Dietzia in some oil production wells significantly increased in the initial phase of nutrient injection and sharply decreased along with the continuous nutrient injection. Overall, this study indicates that Dietzia sp. strain has application potential for enhancing oil recovery through an ex situ way, yet the ability of oil recovery in situ based on nutrient injection is limited.