Strain AS1 Research Articles

Growth and Cell Size of Microalga Auxenochlorella protothecoides AS-1 under Different Trophic Modes.

Certain microalgal species can grow with different trophic strategies depending on the availability of nutrient resources. They can use the energy from light or an organic substrate, or both, and can therefore be called autotrophs, heterotrophs, or mixotrophs. We recently isolated a microalgal strain from the microplastic biofilm, which was identified as Auxenochlorella protothecoides, AS-1. Strain AS-1 grew rapidly in bacterial culture media and exhibited different growth rates and cell sizes under different trophic conditions. We compared the growth performance of AS-1 under the three different trophic modes. AS-1 reached a high biomass (>4 g/L) in 6 days under mixotrophic growth conditions with a few organic carbons as a substrate. In contrast, poor autotrophic growth was observed for AS-1. Different cell sizes, including daughter and mother cells, were observed under the different growth modes. We applied a Coulter Counter to measure the size distribution patterns of AS-1 under different trophic modes. We showed that the cell size distribution of AS-1 was affected by different growth modes. Compared to the auto-, hetero- and mixotrophic modes, AS-1 achieved higher biomass productivity by increasing cell number and cell size in the presence of organic substrate. The mechanisms and advantages of having more mother cells with organic substrates are still unclear and warrant further investigations. The work here provides the growth information of a newly isolated A. protothecoides AS-1 which will be beneficial to future downstream applications.

Entomopathogenic fungi with biological control potential against poultry red mite (Dermanyssus gallinae, Arachnida: Dermanyssidae)

The poultry red mite, Dermanyssus gallinae (Arachnida: Dermanyssidae) is a pest that causes significant economic loss in laying hens for which control methods are limited. In this study, the effects of 20 indigenous fungal strains on poultry red mites in chicken farms were investigated. All experiments were conducted under laboratory condition at 28 ± 1 °C and 80 ± 5% humidity. A screening test showed that Metharizium flavoviride strain As-2 and Beauveria bassiana strain Pa4 had the greatest measured effect on D. gallinae at 1 × 107 conidia/ml 7 days after application. In a subsequent does-response experiment, these strains also caused 92.7% mortality at 1 × 109 conidia/ml within the same period. The LC50 of these strains was 5.5 × 104 (95% CI: 0.8–37.5) conidia/ml for As-2 and 3.2 × 104 (95% CI: 0.4–26.0) conidia/ml for Pa4, and their LT50 were 1.94 and 1.57 days, respectively. The commercial Metarhizium anisopliae bioinsecticide Bio-Storm 1.15% WP, used as a comparator, had LC50 and LT50 1 × 105 (95% CI: 0.1–7.9) conidia/ml and 3.03 (95% CI: 2.4–3.8) days, respectively. It is suggested that mycoacaricides could be developed using the best two fungal strains found in this study (As-2 and Pa4), providing potential for biological control of poultry red mites.

In vitro assessment of the nematicidal potential of Streptomyces violascens strain AS2 against root-knot nematodes (Meloidogyne sp.)

Root-knot nematodes (RKNs, Meloidogyne spp.) present a major challenge to sustainable production. The detrimental environmental impacts and increasing resistance associated with synthetic nematicides have increased the need for biocontrol alternatives. Here, we report for the first time the nematicidal properties of Streptomyces violascens and its viability as a safe and effective solution against nematode infestation. In vitro assays showed that aqueous and ethyl acetate extracts of S. violascens significantly impeded Meloidogyne sp?s reproductive and locomotive functions. The aqueous extract displayed remarkable efficacy, completely inhibiting egg hatching at a concentration of 600 ?g/mL, with an LC50 of 168.38 ?g/mL, and causing 99% immobility of second-stage juveniles (J2s) at 800 ?g/mL within 72 h. Conversely, the organic extract exhibited a 29.3% hatching rate (LC50 of 849.26 ?g/mL) and 79% immobility of J2s under comparable conditions. These findings point to the robust nematicidal potential of S. violascens extracts. Enzymatic profiles supported these results, with the selected isolate producing chitinase and protease enzymes capable of degrading nematode eggshells. Our results show that the S. violascens strain can be integrated into a sustainable nematode management practice as an important agent of RKN biocontrol.

Isolation and characterization of the aspartame-degrading strain Pseudarthrobacter sp. AS-1

Aspartame is one of the main varieties of artificial sweeteners. Although it has been approved as a food additive, the environmental hazards and ecological risks posed by aspartame are attracting more and more attention. In the present study, strain Pseudarthrobacter sp. AS-1 was isolated and characterized as an efficient aspartame degrader. Strain AS-1 was capable of degrading 200 mg L−1 aspartame within 10 h under conditions optimized at 30 °C and pH 8.0. At the same time, it was found that enzymes degrading aspartame in strain AS-1 were induced and secreted extracellularly. Degradation of aspartame in Pseudarthrobacter sp. AS-1 was identified as following: it was first demethylated to aspartyl-phenylalanine, then degraded to phenylalanine and aspartate, and finally the two amino acids were further degraded. In addition, strain AS-1 was able to remove more than 85% of aspartame in soil and river water. It is the first time that pure bacterial cultures were reported to have the capability of aspartame degradation. These findings add to our knowledge of the microbial metabolic mechanisms of aspartame.

Engineered salt-tolerant yeast to improve the physicochemical properties and volatiles of sweet flour paste

Sweet flour paste is a traditional condiment and fermented in high brine solution (14∼18%). Aroma-producing yeast plays a crucial role in the flavor and taste of sweet flour paste, but high salt concentrations often inhibit the action of the yeast. In this study, yeast Zygosaccharomyces rouxii AS2.18 was used as the parent strain to breed a salt-tolerant aroma-producing yeast strain by a combination of atmospheric and room temperature plasma (ARTP) mutagenesis and adaptive laboratory evolution (ALE). The engineered strain AMS79 showed the good growth and fermentation capacity at 18% NaCl concentration. The salt tolerance was closely related to intracellular glycerol production, Na+K+-ATPase activity enhancement and cell wall integrity improvement of the strain. During sweet flour paste fermentation, the maximum amino nitrogen content of the AMS79 group was 1.07- and 1.11-fold higher than that of the AS2.18 group and blank group, respectively. Compared to Blank group, the inoculation of AS2.18 enhanced the relative contents of alcohols, acids, ketones and furan(one)s, and the content of all 7 volatile compound types increased when AMS79 was inoculated for fermentation. In particular, the contents of 2-methyl-butanoic acid, benzeneacetaldehyde, 5-methyl-2-phenyl-2-hexenal and furfural were significantly enhanced by the salt-tolerant strain AMS79 compared to the parent strain AS2.18.

Mutations in asparaginase II from E. coli and implications for inactivation and PEGylation

All clinically-used asparaginases convert L-asparagine (L-Asn) to l-aspartate (L-Asp) and l-glutamine (L-Gln) to L-glutamate (L-Glu), which has been useful in reducing bioavailable asparagine and glutamine in patients under treatment for acute lymphoblastic leukemia. The E. coli type 2 L-asparaginase (EcA2) can present different sequences among varying bacterial strains, which we hypothesized that might affect their biological function, stability and interchangeability. Here we report the analysis of two EcA2 provided by the public health system of a middle-income country. These enzymes were reported to have similar specific activity in vitro, whereas they differ in vivo. Protein sequencing by LC-MS-MS and peptide mapping by MALDI-ToF-MS of their tryptic digests revealed that Aginasa™ share similar sequence to EcA2 from E. coli strain BL21(DE3), while Leuginase™ has sequence equivalent to EcA2 from E. coli strain AS1.357. The two amino acid differences between Aginasa™ (64D and 252 T) and Leuginase™ (64 N and 252S) resulted in structural divergences in solution as accessed by small-angle X-ray scattering and molecular dynamics simulation trajectories. The conformational variability further results in dissimilar surface accessibility with major consequences for PEGylation, as well as different susceptibility to degradation by limited proteolysis. The present results reveal that the sequence variations between these two EcA2 variants results in conformational changes associated with differential conformational plasticity, potentially affecting physico-chemical and biological properties, including proteolytic and immunogenic silent inactivation.

Long-term stability of reactor microbiome through bioaugmentation with Alcaligenes aquatilis AS1 promotes nitrogen removal of piggery wastewater

Bioaugmentation is considered as an attractive method for nitrogen removal in water treatment, but its effectiveness in actual high-strength piggery wastewater has not been adequately verified and the mechanism of bioaugmentation in actual wastewater treatment system is not very clear especially from the perspectives of microbial communities and functional genes. This study investigated the mechanisms of a heterotrophic nitrifying-aerobic denitrifying strain Alcaligenes aquatilis AS1 in the bioaugmentation of continuous biological nitrogen removal of actual piggery wastewater at laboratory scale. The addition of strain AS1 significantly improved the nitrogen removal efficiency (more than 95% of NH4+-N and 75% of TN were removed) and raised the activated sludge resistance to shock loading. AS1 addition also significantly shifted the microbiota structure and interactions among microbial networks were enhanced to obtain the stable bacterial communities. Moreover, strain AS1 achieved effective proliferation and long-term colonization in activated sludge with a relative abundance of genus Alcaligenes more than 70% during the whole operation process and played a dominant role in biological nitrogen removal, while different genera were respectively enriched and involved in pollutants removal at different stages in the control group. In addition, the abundances of most functional genes involved in carbon (C) degradation, carbon fixation and nitrogen (N), phosphorus (P), sulfur (S) cycling in activated sludge were significantly increased in reactor AS1, indicating that strain AS1 not only relied on its unique C and N metabolic activities, but also recruited microorganisms with diverse functions to jointly remove pollutants in wastewater, which could be a common bioaugmentation mechanism in open reactors. This study proves the promising application prospect of strain AS1 in the treatment of high-strength piggery wastewater and shows great importance for guiding bioaugmentation application of functional strains in practical wastewater treatment systems.

Genomic analysis of heavy metal-resistant Halobacterium salinarum isolated from Sfax solar saltern sediments

The draft genome sequences of five archaeal strains, isolated from Sfax solar saltern sediments and affiliated with Halobacterium salinarum, were analyzed in order to reveal their adaptive strategies to live in hypersaline environments polluted with heavy metals. The genomes of the strains (named AS1, AS2, AS8, AS11, and AS19) are found to contain 2,060,688; 2,467,461; 2,236,624; 2,432,692; and 2,428,727 bp respectively, with a G + C content of 65.5, 66.0, 67.0, and 66.2%. The majority of these genes (43.69–55.65%) are annotated as hypothetical proteins. Growth under osmotic stress is possible by genes coding for potassium uptake, sodium efflux, and kinases, as well as stress proteins, DNA repair systems, and proteasomal components. These strains harbor many genes responsible for metal transport/resistance, such as: copper-translocating P-type ATPases, ABC transporter, and cobalt-zinc-cadmium resistance protein. In addition, detoxification enzymes and secondary metabolites are also identified. The results show strain AS1, as compared to the other strains, is more adapted to heavy metals and may be used in the bioremediation of multi-metal contaminated environments. This study highlights the presence of several commercially valuable bioproducts (carotenoids, retinal proteins, exopolysaccharide, stress proteins, squalene, and siderophores) and enzymes (protease, sulfatase, phosphatase, phosphoesterase, and chitinase) that can be used in many industrial applications.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00792-022-01273-0.

Characterization of Alcaligenes aquatilis as a novel member of heterotrophic nitrifier-aerobic denitrifier and its performance in treating piggery wastewater

A novel strain AS1 with heterotrophic nitrifying-aerobic denitrifying capacity in the species of Alcaligenes aquatilis was isolated from the aerobic activated sludge. It showed a great capability of ammonia removal, and the aerobic metabolic pathways to yield gaseous-nitrogen by hydroxylamine oxidation and nitrite denitrification were proposed. AS1 could efficiently remove ammonia under a wide range of environmental conditions, including the ratio of chemical oxygen demand to total nitrogen: 15–30, pH: 6–10, NaCl: 0–60 g/L, shaking speed of 0–180 rpm, and succinate, acetate, or citrate as carbon source. In the treatment of actual piggery wastewater, 95.3%, 95.1% and 84.9% of NH4+-N was removed by AS1 when the initial ammonia concentration was 500, 1300, and 2000 mg/L, respectively, with the maximum NH4+-N removal rate of 30.5 mg/L/h and 569.7 mg/L/d. Furthermore, plate colony-counting showed that AS1 achieved an efficient proliferation. These results imply the application potential of AS1 in treating high-ammonia wastewater.

Antimony transformation and mobilization from stibnite by an antimonite oxidizing bacterium Bosea sp. AS-1

Soils and waters are heavily contaminated by antimony in Xikuangshan (XKS) mine area. It is widely accepted that oxidative dissolution of sulfide minerals and aqueous dissolution are the most prevalent geochemical mechanisms for the release of Sb to the environment. Bosea sp. AS-1 is an antimonite-oxidizer isolated from the mine slag in Xikuangshan Sb mine. Whole genome sequencing revealed the presence of multiple sulfur-oxidizing genes, antimony (Sb) metabolism genes and carbon fixation genes in AS-1′s genome. We therefore hypothesized that under oxic conditions, AS-1 could mediate the oxidation of sulfide and Sb(III) in stibnite (Sb2S3) and lead to the release of Sb. Indeed, strain AS-1 was discovered as an autotrophic Sb(III)-oxidizer. Antimony mobilization studies conducted with strain AS-1 showed significantly enhanced mobilization of Sb, and complete oxidation of released Sb and sulfur to Sb(V) and sulfate. In addition, AS-1 induced a faster release of Sb under heterotrophic condition, and new acicular minerals might form. These findings support the hypothesis that microorganisms play an important role in the mobilization and transformation of Sb in XKS mine area and may contribute to our further understanding of the Sb biogeochemical redox cycle in natural environment.

Proteomics and transcriptomic analysis of Micrococcus luteus strain AS2 under arsenite stress and its potential role in arsenic removal.

The proteomics and transcriptomic analysis of Micrococcus luteus strain AS2 was carried out through 2D gel electrophoresis and RT-PCR. Seven protein spots were selected randomly from the gel and identified through mass spectrometry. Four proteins including putative metal-dependent hydrolase TatD, thioredoxin reductase, DNA-directed RNA polymerase subunit alpha and chaperone protein DnaK were upregulated while superoxide dismutase [Mn], 3-oxoacyl-[acyl-carrier-protein] reductase FabG, and putative alkyl/aryl-sulfatase YjcS were down-regulated under arsenite stress. No significant difference was observed in aioB gene expression analysis in the presence and absence of arsenite. The optimum arsenite processing ability was determined at 37°C (90%) and at pH 7 (92%). The maximum metal processing ability was determined at 250mM arsenite/L (90%) while the minimum was estimated at 1250mM arsenite/L (42%). The maximum arsenite removal ability of strain AS2 determined after 8 days was 68 and 82% from wastewater and distilled water, and the organism can be a good bioresource for green chemistry to eradicate environmental arsenite.

Draft Genome Sequence of Enterobacter sp. AS-1, a Potential Eurytrophic Recombination Host.

Strain AS-1 was isolated from laboratory-scale activated sludge collected in Japan.This strain not only grows on rich medium, including R2A medium, but also forms colonies on medium lacking organic matter other than agar (water agar), indicating it could be used as a eurytrophic recombinant host in material production processes. Here, we present a draft genome sequence of Enterobacter sp. AS-1, which consists of a total of 24 contigs containing 5,207,146 bp, with a GC content of 55.64%, and comprising 4,921 predicted coding sequences. Based on 16S rRNA gene sequence analysis, strain AS-1 was designated as Enterobacter sp. AS-1.

Bio-electrokinetic remediation of crude oil contaminated soil enhanced by bacterial biosurfactant

The present study evaluating the coupling between bioremediation (BIO) and electrokinetic (EK) remediation of crude oil hydrocarbon by using bio-electrokinetic (BIO-EK) technique. The application of bacterial biosurfactant (BS) may increase the remediation efficiency by increasing the solubility of organic materials. In this work, the potential biosurfactant producing marine bacteria were isolated and identified by 16S rDNA analysis namely Bacillus subtilis AS2, Bacillus licheniformis AS3 and Bacillus velezensis AS4. Biodegradation efficiency of crude oil was found as 88%, 92% and 97% for strain AS2, AS3 and AS4 respectively, with the optimum temperature of 37 °C and pH 7. FTIR confirm the BS belongs to lipopeptide in nature. GCMS reveals that three isolates degraded the lower to higher molecular weight of the crude oil (C8 to C28) effectively. Results showed that use of BS in electokinetic remediation enhance the biodegradation rate of crude oil contaminated soil about 92% than EK (60%) in 2 days operation. BS enhances the solubilization of hydrocarbon and it leads to the faster electromigration of hydrocarbon to the anodic compartment, which was confirmed by the presence of higher total organic content than the EK. This study proven that the BIO-EK combined with BS can be used to enhance in situ bioremediation of petroleum contaminated soils.

Efficacy of metabolites of a Streptomyces strain (AS1) to control growth and mycotoxin production by Penicillium verrucosum, Fusarium verticillioides and Aspergillus fumigatus in culture

The objectives of this study were to determine the efficacy of metabolites of a Streptomyces strain AS1 on (a) spore germination, (b) mycelial growth, (c) control of mycotoxins produced by Penicillium verrucosum (ochratoxin A, OTA), Fusarium verticillioides (fumonisins, FUMs) and Aspergillus fumigatus (gliotoxin) and (d) identify the predominant metabolites involved in control. Initial screening showed that the Streptomyces AS1 strain was able to inhibit the mycelial growth of the three species at a distance, due to the release of secondary metabolites. A macroscopic screening system showed that the overall Index of Dominance against all three toxigenic fungi was inhibition at a distance. Subsequent studies showed that the metabolite mixture from the Streptomyces AS1 strain was very effective at inhibiting conidial germination of P. verrucosum, but less so against conidia of A. fumigatus and F. verticillioides. The efficacy was confirmed in studies on a conducive semi-solid YES medium in BioScreen C assays. Using the BioScreen C and the criteria of Time to Detection (TTD) at an OD = 0.1 showed good efficacy against P. verrucosum when treated with the Streptomyces AS1 extract at 0.95 and 0.99 water activity (aw) when compared to the other two species tested, indicating good efficacy. The effective dose for 50% control of growth (ED50) at 0.95 and 0.99 aw were approx. 0.005 ng/ml and 0.15 μg/ml, respectively, with the minimum inhibitory concentration (MIC) at both aw levels requiring > 40 μg/ml. In addition, OTA production was completely inhibited by 2.5 μg/ml AS1 extract at both aw levels in the in vitro assays. Ten metabolites were identified with four of these being predominant in concentrations > 2 μg/g dry weight biomass. These were identified as valinomycin, cyclo(L-Pro-L-Tyr), cyclo(L-Pro-L-Val) and brevianamide F.

Phenotypic and genomic analysis of multiple heavy metal-resistant Micrococcus luteus strain AS2 isolated from industrial waste water and its potential use in arsenic bioremediation.

Multiple heavy metal-resistant bacterium, Micrococcus luteus strain AS2, was isolated from industrial waste water of District Sheikhupura, Pakistan. The isolated bacterium showed minimum inhibitory concentrations of 55 and 275mM against arsenite and arsenate. The bacterial strain also showed resistance against other heavy metal ions, i.e., lead, cadmium, chromium, mercury, nickel, and zinc, apart from arsenic. The optimum temperature and pH were 37°C and 7, respectively. The antioxidant enzymes such as catalase were significantly increased under arsenite stress. The increase in 43.9% of GSH/GSSG and 72.72% of non-protein thiol was determined under15mM arsenite stress. Bacterial genome was sequenced through Illumina and Nanopore and genes related to arsenic and other heavy metals were identified and blast (tblastx) on NCBI. Through scanning electron microscopy, no morphological changes were observed in bacterial cells under arsenite stress. The peaks appeared in EDX showed that there is surface adsorption of arsenite in bacterial cell while it was confirmed from Fourier transformed infrared spectroscopy analysis that there is some interaction between arsenite and functional groups present on the surface of bacterial cell. The SDS-PAGE analysis of whole-cell proteins under 15mM arsenite stress clearly revealed that there is upregulation of some proteins in ranged of 60 to 34kDa. The bioremediation efficiency (E) of bacterial biomass was 72% after 2h and 99% after 10h. The bioremediation efficiency of bacterial biomass is an indicator for the isolated bacterium to employ as a potential candidate for the amelioration of sites contaminated with arsenic.

Culture-dependent hunt and characterization of iron-oxidizing bacteria in Baiyin Copper Mine, China, and their application in metals extraction.

The exploration of microbial diversity in extremely acidic habitats has provided a vital base for the progression of minerals biotechnology. Three indigenous iron-oxidizing acidophilic bacterial strains were isolated through serial dilution of enriched bacterial culture from Baiyin Copper Mine Stope, China. The morphological, biochemical, physiological, and phylogenetic characteristics of isolates were investigated. These isolates were motile, Gram-negative, and curved shape with pleomorphism except isolate WG101 that was a straight rod. The optimum growth pH and temperature for all isolates were 1.5 and 30 °C, respectively, and showed extreme acidophilic nature. All the isolates showed obligate chemoautotrophic nature and used ferrous iron and pyrite as an energy source, however, isolates WG102 and WG103 were unable to use sulfur, while isolate WG101 could use elemental sulfur and reduced inorganic sodium thiosulfate as an energy source. The phylogenetic analyses based on 16S rRNA sequences revealed that the isolates WG101, WG102, and WG103 were homologous with Acidithiobacillus ferrooxidans strain AS2 (99%), Leptospirillum ferriphilum strain YSK (98%), and Leptospirillum ferrooxidans strain L15 (98%), respectively. These bacterial isolates showed efficient copper and zinc dissolution from the ore. The metals dissolution rate of At. ferrooxidans strain WG101 was 54.5 ± 4.33% (copper) and 49.6 ± 5% (zinc). The metals recovery rate of L. ferriphilum strain WG102 was 45.7 ± 3.5% (copper) and 40.5 ± 2.5% (zinc). The recovery rate of copper and zinc was 49.6 ± 4% and 46.5 ± 3% respectively in the case of L. ferrooxidans strain WG103. The findings of this study are consistent with the notion that the indigenous bacteria are more efficient in minerals dissolution.

Evaluation of butachlor biodegradation efficacy of Serratia ureilytica strain AS1: a statistical optimization approach

Butachlor is a chloroacetanilide class of herbicide, commonly administered to control unwanted grasses and broad leaf weeds. Extensive usage of the herbicide has led to the contamination of water bodies and surrounding areas, resulting in an adverse impact on the environment. In the present work, a novel butachlor-catabolizing bacterium Serratia ureilytica strain AS-1 was isolated from an herbicide-contaminated soil. Statistical optimization techniques were used to optimize the butachlor biodegradation. Experimental parameters such as growth temperature, pH of the medium and biomass concentration were found to be significant for butachlor biodegradation. The results obtained indicates that the maximum degradation of 2.08 mg/L/h of butachlor was achieved under the optimal conditions of 32.5 °C of incubation temperature, pH 7.5 and 10% (v/v) inoculum size along with a polynomial mathematical model having R2 = 0.9833. The model was corroborated by carrying out experiments at the optimized conditions.

Characterization of the antimonite- and arsenite-oxidizing bacterium Bosea sp. AS-1 and its potential application in arsenic removal

Arsenic (As) and antinomy (Sb) usually coexist in natural environments where both of them pollute soils and water. Microorganisms that oxidize arsenite [As(III)] and tolerate Sb have great potential in As and Sb bioremediation, In this study, a Gram-negative bacterial strain, Bosea sp. AS-1, was isolated from a mine slag sample collected in Xikuangshan Sb mine in China. AS-1 could tolerate 120 mM of As(III) and 50 mM of antimonite [Sb(III)]. It could also oxidize 2 mM of As(III) or Sb(III) completely under heterotrophic and aerobic conditions. Interestingly, strain AS-1 preferred to oxidize As(III) with yeast extract as the carbon source, whereas Sb(III) oxidation was favored with lactate in the medium. Genomic analysis of AS-1 confirmed the presence of several gene islands for As resistance and oxidation. Notably, a system of AS-1 and goethite was found to be able to remove 99% of the As with the initial concentration of 500 μg/L As(III) and 500 μg/L Sb(III), which suggests the potential of this approach for As removal in environments especially with the presence of high Sb.

Bioleaching of copper- and zinc-bearing ore using consortia of indigenous iron-oxidizing bacteria.

Indigenous iron-oxidizing bacteria were isolated on modified selective 9KFe2+ medium from Baiyin copper mine stope, China. Three distinct acidophilic bacteria were isolated and identified by analyzing the sequences of 16S rRNA gene. Based on published sequences of 16S rRNA gene in the GenBank, a phylogenetic tree was constructed. The sequence of isolate WG101 showed 99% homology with Acidithiobacillus ferrooxidans strain AS2. Isolate WG102 exhibited 98% similarity with Leptospirillum ferriphilum strain YSK. Similarly, isolate WG103 showed 98% similarity with Leptospirillum ferrooxidans strain L15. Furthermore, the biotechnological potential of these isolates in consortia form was evaluated to recover copper and zinc from their ore. Under optimized conditions, 77.68 ± 3.55% of copper and 70.58 ± 3.77% of zinc were dissolved. During the bioleaching process, analytical study of pH and oxidation-reduction potential fluctuations were monitored that reflected efficient activity of the bacterial consortia. The FTIR analysis confirmed the variation in bands after treatment with consortia. The impact of consortia on iron speciation within bioleached ore was analyzed using Mössbauer spectroscopy and clear changes in iron speciation was reported. The use of indigenous bacterial consortia is more efficient compared to pure inoculum. This study provided the basic essential conditions for further upscaling bioleaching application for metal extraction.

Atomic Force Microscopy of Side Wall and Septa Peptidoglycan From Bacillus subtilis Reveals an Architectural Remodeling During Growth.

Peptidoglycan is the fundamental structural constituent of the bacterial cell wall. Despite many years of research, the architecture of peptidoglycan is still largely elusive. Here, we report the high-resolution architecture of peptidoglycan from the model Gram-positive bacterium Bacillus subtilis. We provide high-resolution evidence of peptidoglycan architecture remodeling at different growth stages. Side wall peptidoglycan from B. subtilis strain AS1.398 changed from an irregular architecture in exponential growth phase to an ordered cable-like architecture in stationary phase. Thickness of side wall peptidoglycan was found to be related with growth stages, with a slight increase after transition to stationary phase. Septal disks were synthesized progressively toward the center, while the surface features were less clear than those imaged with side walls. Compared with previous studies, our results revealed slight differences in architecture of peptidoglycan from different B. subtilis strains, expanding our knowledge about the architectural features of B. subtilis peptidoglycan.