Strain Enterobacter Sp Research Articles

Production of ethanol by Enterobacter sp. EtK3 during fruit waste biotransformation

Ethanol resistant bacterial strain Enterobacter sp. EtK3 was isolated from kitchen wastewater through enrichment culture. The isolated strain was found to produce ethanol by fermenting dried banana peel without any pre-treatment. Ethanol production was confirmed by High-Performance Liquid Chromatography (HPLC) analysis of the fermentation broth. Sequence analysis of the 16S rRNA gene of bacterial isolate EtK3 confirmed its relation with the Enterobacteriaceae family. The temperature of 35 °C with pH 7 utilizing 15.5 gL-1 fruit waste was found to be most favorable for ethanol production after a two-level, three-factor optimization study using Response Surface Methodology (RSM). In the optimized condition, a 23.6% yield of ethanol was obtained. Kinetic study of cell growth, ethanol production and reducing sugar utilization revealed that ethanol production was growth associated with product inhibition at the late log phase. Proximate analysis of pre and post-fermented fruit waste showed subsequent depletion of the fixed carbon content of waste indicating carbon utilization of EtK3 strain from the waste sample. Waste degradation was further confirmed by Scanning Electron Microscope with Energy Dispersive X-Ray (SEM-EDX) and Fourier Transform Infrared Spectrometer (FTIR). An empirical representation of the waste was derived to produce a complete mass balance of the whole ethanol production process from fruit waste.

Engineering banana endosphere microbiome to improve Fusarium wilt resistance in banana

BackgroundPlant microbiome highlights the importance of endosphere microbiome for growth and health of the host plant. Microbial community analysis represents an elegant way to identify keystone microbial species that have a more central position in the community. The aim of this study was to access the interactions between the keystone bacterial species and plants during banana Fusarium wilt process, by comparing the endophytic bacterial and fungal community in banana roots and shoot tips during growth and wilting processes. The keystone bacterial species were isolated and further engineered to improve banana wilt resistance.ResultsBanana endosphere microbiome structure varied during plant growth and wilting processes. Bacterial and fungal diversity in the shoot tips and roots increased with the development of the banana plantlets. The bacterial groups belonging to the Enterobacteriaceae family with different relative abundances were detected in all the samples. The Klebsiella spp. might be the keystone bacteria during the growth of banana plantlets. The relative abundance of Fusarium associated with the wilt disease did not increase during the wilting process. The endophytic Enterobacteriaceae strains Enterobacter sp. E5, Kosakonia sp. S1, and Klebsiella sp. Kb were isolated on Enterobacteriaceae selective medium and further engineered by expressing 1-aminocyclopropane-1-carboxylate (ACC) deaminase on the bacterial cell walls (designated as E5P, S1P, and KbP, respectively). Pot experiments suggested that plants inoculated with strains E5, E5P, S1, and S1P increased resistance to the Fusarium wilt disease compared with the controls without inoculation, whereas the Klebsiella inoculation (Kb and KbP) did not increase the wilt resistance. Compared with the inoculation with the wild strains E5 and S1, the inoculation with engineered strains E5P and S1P significantly increased wilt resistance and promoted plant growth, respectively. The results illustrated that the keystone species in the banana microbiome may not be dominant in numbers and the functional role of keystone species should be involved in the wilt resistance.ConclusionThe ACC deaminase activity of engineered bacteria was essential to the Fusarium wilt resistance and growth promotion of banana plants. Engineering keystone bacteria in plant microbiome with ACC deaminase on the cell walls should be a promising method to improve plant growth and disease resistance.

Enterobacter sp. strain Fs-11 adapted to diverse ecological conditions and promoted sunflower achene yield, nutrient uptake, and oil contents.

Plant growth-promoting rhizobacteria are under extensive investigation to supplement the chemical fertilizers due to cost-effective and eco-friendly nature. However, their consistency in heterogeneous soil and diverse ecological settings is unclear. The current study presents in vitro and field evaluation of pre-characterized PGPR strain Enterobacter sp. Fs-11 (GenBank accession # GQ179978) in terms of its potential to enhance sunflower yield and oil contents under diverse environmental conditions. Under in vitro conditions, strain Fs-11 showed optimal growth at a range of temperature (15 to 40°C) and pH values (6.5 to 8.5). Extracellular and intracellular localizations of the strain Fs-11 in sunflower root cortical cells through transmission electron microscopy confirmed its epiphytic and endophytic colonization patterns, respectively. In field experiments, conducted at three different agro-climatic locations, inoculation of strain Fs-11 at 50% reduced NP fertilizer resulted in a significant increase in growth, achene yield, nutrient uptake, and oil contents. Inoculation also responded significantly in terms of increase in mono- and polyunsaturated fatty acids (oleic and linoleic acids, respectively) without rising saturated fatty acid (palmitic and stearic acids) contents. We concluded that Enterobacter sp. Fs-11 is a potential candidate for biofertilizer formulations to supplement chemical fertilizer requirements of sunflower crop under diverse climatic conditions.

Production of bacterial cellulose hydrogels with tailored crystallinity from Enterobacter sp. FY-07 by the controlled expression of colanic acid synthetic genes

Hydrogels exhibit smart three-dimensional networks and extraordinary water-absorbing ability. To improve the water-holding capacity of bacterial cellulose hydrogels, the expression of a biosynthetic gene cluster of colanic acid, a water-soluble polysaccharide, was induced in Enterobacter sp. FY-07, which produces an abundance of bacterial cellulose hydrogel under aerobic and anaerobic fermentation conditions. The results indicated that in situ modified bacterial cellulose hydrogels with different crystallinities, rheological properties and water-holding capacities were produced by cultivating the engineered strain Enterobacter sp. FY-07::tac under different inducing conditions. The water-holding capacity of the modified bacterial cellulose hydrogel was enhanced by more than 1.7 fold compared to the hydrogel produced by Enterobacter sp. FY-07, and the networks of the modified bacterial cellulose hydrogel were densified but still clear. These results suggest that this in situ modification strategy endows bacterial cellulose hydrogels with improved properties and potentially expands their applications in biomedical fields and the food industry.

Genome mining of 2-phenylethanol biosynthetic genes from Enterobacter sp. CGMCC 5087 and heterologous overproduction in Escherichia coli

Background2-Phenylethanol (2-PE) is a higher aromatic alcohol that is widely used in the perfumery, cosmetics, and food industries and is also a potentially valuable next-generation biofuel. In our previous study, a new strain Enterobacter sp. CGMCC 5087 was isolated to produce 2-PE from glucose through the phenylpyruvate pathway.ResultsIn this study, candidate genes for 2-PE biosynthesis were identified from Enterobacter sp. CGMCC 5087 by draft whole-genome sequence, metabolic engineering, and shake flask fermentation. Subsequently, the identified genes encoding the 2-keto acid decarboxylase (Kdc) and alcohol dehydrogenase (Adh) enzymes from Enterobacter sp. CGMCC 5087 were introduced into E. coli BL21(DE3) to construct a high-efficiency microbial cell factory for 2-PE production using the prokaryotic phenylpyruvate pathway. The enzymes Kdc4427 and Adh4428 from Enterobacter sp. CGMCC 5087 showed higher performances than did the corresponding enzymes ARO10 and ADH2 from Saccharomyces cerevisiae, respectively. The E. coli cell factory was further improved by overexpressing two upstream shikimate pathway genes, aroF/aroG/aroH and pheA, to enhance the metabolic flux of the phenylpyruvate pathway, which resulted in 2-PE production of 260 mg/L. The combined overexpression of tktA and ppsA increased the precursor supply of erythrose-4-phosphate and phosphoenolpyruvate, which resulted in 2-PE production of 320 mg/L, with a productivity of 13.3 mg/L/h.ConclusionsThe present study achieved the highest titer of de novo 2-PE production of in a recombinant E. coli system. This study describes a new, efficient 2-PE producer that lays foundation for the industrial-scale production of 2-PE and its derivatives in the future.

Genome sequence of the endophytic strain Enterobacter sp. J49, a potential biofertilizer for peanut and maize

Enterobacter sp. J49 is a plant growth promoting endophytic strain that promotes the growth of peanut and maize crops. This strain promotes plant growth by different mechanisms with the supply of soluble phosphorus being one of the most important. Enterobacter sp. J49 not only increases the phosphorus content in the plant but also in the soil favoring the nutrition of other plants usually used in rotation with these crops. The aims of this study were to analyze the genome sequence of Enterobacter sp. J49 in order to deepen our knowledge regarding its plant growth promoting traits and to establish its phylogenetic relationship with other species of Enterobacter genus. Genome sequence of Enterobacter sp. J49 is a valuable source of information to continuing the research of its potential industrial production as a biofertilizer of peanut, maize and other economically important crops.

Exopolysaccharide produced by Enterobacter sp. YG4 reduces uranium induced nephrotoxicity

Uranium nephrotoxicity is a health concern with very few treatment options. Bacterial exopolysaccharides (EPS) possess multiple biological activities and appear as prospective candidates for treating uranium nephrotoxicity. This study focuses on the ability of an EPS produced by a bacterial strain Enterobacter sp. YG4 to reduce uranium nephrotoxicity in vivo. This bacterium was isolated from the gut contents of a slug Laevicaulis alte (Férussac). Based on the aniline blue staining reaction and infrared spectral analysis, the EPS was identified as β-glucan and its molecular weight was 11.99×106Da. The EPS showed hydroxyl radical scavenging ability and total antioxidant capacity in vitro. To assess the protection provided by the EPS against uranium nephrotoxicity, a single dose of 2mg/kg uranyl nitrate was injected intraperitoneally to albino Wistar rats. As intervention, the EPS was administered orally (100mg/kg/day) for 4 consecutive days. The rats were sacrificed on the fifth day and analyses were conducted. Increased serum creatinine and urea nitrogen levels and histopathological alterations in kidneys were observed in uranyl nitrate treated animals. All these alterations were reduced with the administration of Enterobacter sp. YG4 EPS, emphasizing a novel approach in treating uranium nephrotoxicity.

Biochemical Basis of Mercury Remediation and Bioaccumulation by Enterobacter sp. EMB21

The aims of this study were to isolate metal bioaccumulating bacterial strains and to study their applications in removal of environmental problematic heavy metals like mercury. Five bacterial strains belonging to genera Enterobacter, Bacillus, and Pseudomonas were isolated from oil-spilled soil. Among these, one of the strains Enterobacter sp. EMB21 showed mercury bioaccumulation inside the cells simultaneous to its bioremediation. The bioaccumulation of remediated mercury was confirmed by transmission electron microscopy and energy dispersive X-ray. The mercury-resistant loci in the Enterobacter sp. EMB21 cells were plasmid-mediated as confirmed by transformation of mercury-sensitive Escherichia coli DH5α by Enterobacter sp. EMB21 plasmid. Effect of different culture parameters viz-a-viz inoculum size, pH, carbon, and nitrogen source revealed that alkaline pH and presence of dextrose and yeast extract favored better remediation. The results indicated the usefulness of Enterobacter sp. EMB21 for the effective remediation of mercury in bioaccumulated form. The Enterobacter sp. EMB21 seems promising for heavy metal remediation wherein the remediated metal can be trapped inside the cells. The process can further be developed for the synthesis of valuable high-end functional alloy, nanoparticles, or metal conjugates from the metal being remediated.

Target-oriented discovery of a new esterase-producing strain Enterobacter sp. ECU1107 for whole cell-catalyzed production of (2S,3R)-3-phenylglycidate as a chiral synthon of Taxol

A new strain, Enterobacter sp. ECU1107, was identified among over 200 soil isolates using a two-step screening strategy for the enantioselective synthesis of (2S,3R)-3-phenylglycidate methyl ester (PGM), a key intermediate for production of a potent anticancer drug Taxol®. An organic-aqueous biphasic system was employed to reduce spontaneous hydrolysis of the substrate PGM and isooctane was found to be the most suitable organic solvent. The temperature and pH optima of the whole cell-mediated bioreaction were 40 °C and 6.0, respectively. Under these reaction conditions, the enantiomeric excess (ee(s)) of (2S,3R)-PGM recovered was greater than 99 % at approximately 50 % conversion. The total substrate loading in batch reaction could reach 600 mM. By using whole cells of Enterobacter sp. ECU1107, (2S,3R)-PGM was successfully prepared in decagram scale in a 1.0-l mechanically stirred reactor, affording the chiral epoxy ester in >99 % ee s and 43.5 % molar yield based on the initial load of racemic substrate.

Biosynthesis of polyhydroxybutyrate in Enterobacter sp. SEL2 and Enterobacteriaceae bacterium sp.PFW1 using sugar cane molasses as media

The present research was conducted to check the cane molasses as a media for the production of polyhydroxybutyrate (PHB). Different dilutions of cane molasses were used in combination with water and mineral medium and in different ratios of carbon and nitrogen sources. Out of the 54 bacteria isolated from three types of organic waste contaminated environments, two were selected for their highest intensity of fluorescence under UV by Nile blue A viable colony staining method and they produced maximum amount of PHB from glucose (66.61±0.05% and 76.92±0.04%) in a shake flask culture at pH 7.0, 37°C and 150 rpm. The bacterial strains were identified as Gram negative rods and the 16S ribosomal ribonucleic acid (RNA) sequence similarity search showed 99 and 98% homology to Enterobacter sp. andEnterobacteriaceae bacterium respectively. In bacterial strains Enterobacter sp. SEL2 (JF901810) and Enterobacteriaceae bacterium PFW1 (JF901811), the polyhydroxybutyrate (PHB) biosynthesis from molasses was observed as 57.61±0.57% and 58.07±0.25% respectively at pH 7.0, 37°C and 150 rpm. The optimal time was 24 to 48 h for strain SEL2 and 48 to 72 h for strain PFW1. The best growth and polyhydroxyalkanoates (PHA) production was observed in media with 2% molasses and 0.2% ammonium sulphate in mineral medium (Media 11). The functional groups of the extracted PHA granules were identified as C=O group by Fourier transform infrared (FTIR) spectroscopy analysis and proton nuclear magnetic resonance (NMR) analysis confirmed it as a homopolymer of hydroxybutyrate. Key words: Polyhydroxyalkanoates (PHA), Sudan Black B, Nile blue A, molasses, polyhydroxybutyrate (PHB), Enterobacter, Enterobacteriaceae.

A batch decolorization and kinetic study of Reactive Black 5 by a bacterial strain Enterobacter sp. GY-1

An Enterobacter strain (GY-1) with high activity of decolorization of Reactive Black 5 (RB 5) was isolated from textile wastewater treating sludge. The kinetic characteristics of dye decolorization by the strain GY-1 were determined quantitatively using the diazo dye, RB 5. Effects of different operation parameters (inoculum size, pH, temperature and salinity) and various electron donors on decolorization of the azo dye by GY-1 were systematically investigated to reveal the primary factors that determine the performance of the azo dye decolorization. The decolorization of RB 5 was attributed to extracellular enzymes. A kinetic model was established giving the dependence of decolorization rate on cell mass concentration (first order). Decolorization rate increased with increasing temperature from 20 to 35 °C, which can be predicted by Arrhenius equation with the activation energy ( E a) of 8.50 kcal mol −1 and the frequency factor ( A 0) of 6.28 × 10 7 mg l g MLSS −1 h −1. Michaelis–Menten kinetics and Eadie–Hofstee plot were used to determine V max, 1.05 mg l −1 h −1 and K m, 24.06 mg l −1.

Physiological adaptations and tolerance towards higher concentration of selenite (Se+4) in Enterobacter sp. AR-4, Bacillus sp. AR-6 and Delftia tsuruhatensis AR-7

Environmental contamination with selenium is a major health concern. A few bacterial strains have been isolated that can transform toxic selenite to non-toxic elemental selenium only at low concentrations (0.001-150 mM) in recent past. We have previously reported isolation and characterization of few selenite-tolerant bacterial strains. These strains were found to be resistant to selenite at (300-600 mM) concentrations. In the present study we have characterized some physiological adaptations of strains Enterobacter sp. AR-4, Bacillus sp. AR-6 and Delftia tsuruhatensis AR-7 during exposure to higher concentration of selenite under aerobic and anaerobic environments. Adaptive responses are largely associated with alteration of cell morphology and change in total cellular fatty acid composition. Interestingly, electron microscopy studies revealed substantial decrease in cell size and intracellular deposition of Se(0) crystals when reduction is carried out under aerobic conditions. On the other hand, cell size increased with adhesion of Se(0) on cell surface during anaerobic reduction. Fatty acid composition analysis demonstrated selective increase in saturated and cyclic fatty acids and decrease in unsaturated ones during aerobic transformation. Changes observed during anaerobic transformation were in surprising contrast as indicated by total absence of saturated and cyclic fatty acids. Results presented here provide evidences for putative occurrence of two distinct mechanisms involved in tolerance towards higher concentrations of selenite utilization under aerobic and anaerobic conditions. Further, prior exposure to higher concentration of Se(+4) enabled rapid adaptation indicating role of inducible system in adaptation.

Biological decolorization of the reactive dyes Reactive Black 5 by a novel isolated bacterial strain Enterobacter sp. EC3

Studies were carried out on the decolorization of the reactive dye Reactive Black 5 by a newly isolated bacterium, EC3. Phenotypic characterization and phylogenetic analysis based on 16S rDNA sequence comparisons indicate that this strain belonged to the genus Enterobacter. The optimal conditions for the decolorizing activity of Enterobacter sp . EC3 were anaerobic conditions with glucose supplementation, at pH 7.0, and 37 °C. The maximum decolorization efficiency against Reactive Black 5 achieved in this study was 92.56%. Ultra-violet and visible (UV–vis) analyses before and after decolorization and the colorless bacterial biomass after decolorization suggested that decolorization was due to biodegradation, rather than inactive surface adsorption. The bacterial strain also showed a strong ability to decolorize various reactive textile dyes, including both azo and anthraquinone dyes. To our knowledge, it is the first time that a bacterial strain of Enterobacter sp. has been reported with decolorizing ability against both azo and anthraquinone dyes.

Mouse Single Oral Dose Toxicity Studies of PGB-1, a Novel Polyglucosamine Polymer Produce from Enterobacter sp. BL-2

This study was conducted to obtain acute information of the oral dose toxicity of PGB-1, a novel polyglucosamine polymer produced from a new strain Enterobacter sp. BL-2 in male and female mice. In order to calculated 50% lethal dose (LD??) and approximate lethal dose (LD), test material was once orally administered to male and female ICR mice at dose levels of 2000, 1000, 500, 250, 125 and 0 (vehicle control) ㎖/㎏ (body wt.). The mortality and changes on body weight, clinical signs, gross observation and organ weight and histopathology of principle organs were monitored 14 days after dosing with PGB-1. We could not find any mortalities, clinical signs, body weight changes and gross findings. In addition, significant changes in the organ weight and histopathology of principal organs were not observed except for some sporadic findings. The results obtained in this study suggest that PGB-1 may not be toxic in mice and may be therefore safe for clinical use. The LD?? and approximate LD in mice after single oral dose of PGB-1 were considered over 2000 ㎎/㎏ in both female and male mice.

Acetate-mediated pH-stat fed-batch cultivation of transconjugant Enterobacter sp. BL-2S over-expressing glmS gene for excretive production of microbial polyglucosamine PGB-1

A unique cationic polyglucosamine biopolymer PGB-1 comprising more than 95% D-glucosamine was excretively produced from a new bacterial strain Enterobacter sp. BL-2 under acetate-mediated culture conditions. Since the biopolymer PGB-1 could be synthesized from the UDP-N-acetylglucosamine monomer derived from the hexosamine pathway, three glmS, glmM, and glmU genes in the hexosamine pathway were cloned from Enterobacter sp. BL-2, and their molecular structures were elucidated. The cloned glmS, glmM, and glmU genes were reintroduced into the parent strain Enterobacter sp. BL-2 through a conjugative transformation for the overproduction of the biopolymer PGB-1. The biopolymer production increased 1.5-fold in the transconjugant Enterobacter sp. BL-2S over-expressing the first-step glmS gene encoding glucosamine-6-phosphate synthase. The transconjugant Enterobacter sp. BL-2S was cultivated pH-stat fed-batch widely, while intermittently feeding an acetate solution to maintain a constant pH level of 8.0 for 72 h, resulting in 1.15 g/L of the extracellular polyglucosamine biopolymer PGB-1.