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Lipase is an important commercial enzyme with unique and versatile biotechnological applications. This study was conducted to biosynthesize and characterizes alkaliphilic lipase by Exiguobacterium sp. strain AMBL-20T isolated from the glacial water samples of the northeastern (Gilgit-Baltistan) region of Pakistan. The isolated bacterium was identified as Exiguobaterium sp. strain AMBL-20T on the basis of morphological, biochemical, and phylogenetic analysis of 16S rRNA sequences with GenBank accession number MW229267. The bacterial strain was further screened for its lipolytic activity, biosynthesis, and characterization by different parameters with the aim of maximizing lipase activity. Results showed that 2% Olive oil, 0.2% peptone at 25 °C, pH 8, and 24 h of incubation time found optimal for maximum lipase production. The lipase enzyme was partially purified by ammonium sulphate precipitation and its activity was standardized at pH 8 under 30 °C temperature. The enzyme showed functional stability over a range of temperature and pH. Hence, extracellular alkaliphilic lipase from Exiguobacterium sp. is a potential candidate with extraordinary industrial applications, particularly in bio-detergent formulations.

The Exiguobacterium genus comprises Gram-stain-positive and facultatively anaerobic bacteria. Some Exiguobacterium species have previously shown significant high 16S rRNA gene sequence similarities with each other. This study evaluates the taxonomic classification of those Exiguobacterium species through comprehensive genome analysis. Average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) values were determined for various Exiguobacterium species pairs. The ANI and dDDH values between Exiguobacterium enclense and Exiguobacterium indicum, Exiguobacterium aquaticum and Exiguobacterium mexicanum, Exiguobacterium soli and Exiguobacterium antarcticum, and Exiguobacterium sibiricum and Exiguobacterium artemiae were above the cut-off level (95–96% for ANI and 70% for dDDH) for species delineation. Based on the findings, we propose to reclassify Exiguobacterium enclense as a later heterotypic synonym of Exiguobacterium indicum, Exiguobacterium aquaticum as a later heterotypic synonym of Exiguobacterium mexicanum, Exiguobacterium soli as a later heterotypic synonym of Exiguobacterium antarcticum and Exiguobacterium sibiricum as a later heterotypic synonym of Exiguobacterium artemiae.

The tolerance of plant growth-promoting endophytes (PGPEs) against various concentrations of cadmium (Cd) and nickel (Ni) was investigated. Two glutathione-producing bacterial strains (Enterobacter ludwigii SAK5 and Exiguobacterium indicum SA22) were screened for Cd and Ni accumulation and tolerance in contaminated media, which showed resistance up to 1.0 mM. Both strains were further evaluated by inoculating specific plants with the bacteria for five days prior to heavy metal treatment (0.5 and 1.0 mM). The enhancement of biomass and growth attributes such as the root length, shoot length, root fresh weight, shoot fresh weight, and chlorophyll content were compared between treated inoculated plants and treated non-inoculated plants. Both strains significantly increased the accumulation of Cd and Ni in inoculated plants. The accumulation of both heavy metals was higher in the roots than in the shoots, however; Ni accumulation was greater than Cd. Heavy metal stress-responsive genes such as OsGST, OsMTP1, and OsPCS1 were significantly upregulated in treated non-inoculated plants compared with treated inoculated plants, suggesting that both strains reduced heavy metal stress. Similarly, abscisic acid (ABA) was increased with increased heavy metal concentration; however, it was reduced in inoculated plants compared with non-inoculated plants. Salicylic acid (SA) was found to exert synergistic effects with ABA. The application of suitable endophytic bacteria can protect against heavy metal hyperaccumulation by enhancing detoxification mechanisms.

Plant growth-promoting rhizobacteria (PGPR) have a positive effect on plant development and being a promising way to enhance crop productivity and as substitution of chemical fertilizers. Selenium (Se) is an important trace element and its intake is usually lower than the daily minimum amount required for humans; hence, there is a demand on the design of Se biofortification strategies. Here, the genetic traits known to be associated with Plant-Growth Promotion (PGP) and Se biotransformation of Exiguobacterium sp. S17 were evaluated through genome analysis. Its growth-promoting capacity was tested through plant-growth promotion assays in laboratory and field conditions, using Brassica juncea (indian mustard), Beta vulgaris (chard), and Lactuca sativa (lettuce). Additionally, the Se biotransformation ability of Exiguobacterium sp. S17 was evaluated and the obtained selenized bacteria were tested in mustard plants. The sequenced bacteria genome revealed the presence of multiple genes involved in important functions regarding soil and plant colonization, PGP and Se biotransformation. Moreover, it was demonstrated that Exiguobacterium sp. S17 enhanced plant growth and could be useful to produce Se accumulation and biofortification in accumulator plants such as mustard. Thereby, Exiguobacterium sp. S17 might be used for developing new, sustainable, and environmentally friendly agro-technological strategies.

Background The bacterial genus Exiguobacterium includes several species that inhabit environments with a wide range of temperature, salinity, and pH. This is why the microorganisms from this genus are known generically as polyextremophiles. Several environmental isolates have been explored and characterized for enzyme production as well as for bioremediation purposes. In this line, toxic metal(loid) reduction by these microorganisms represents an approach to decontaminate soluble metal ions via their transformation into less toxic, insoluble derivatives. Microbial-mediated metal(loid) reduction frequently results in the synthesis of nanoscale structures—nanostructures (NS) —. Thus, microorganisms could be used as an ecofriendly way to get NS. Results We analyzed the tolerance of Exiguobacterium acetylicum MF03, E. aurantiacum MF06, and E. profundum MF08 to Silver (I), gold (III), and tellurium (IV) compounds. Specifically, we explored the ability of cell-free extracts from these bacteria to reduce these toxicants and synthesize NS in vitro , both in the presence or absence of oxygen. All isolates exhibited higher tolerance to these toxicants in anaerobiosis. While in the absence of oxygen they showed high tellurite- and silver-reducing activity at pH 9.0, whereas AuCl 4 − which was reduced at pH 7.0 in both conditions. Given these results, cell-free extracts were used to synthesize NS containing silver, gold or tellurium, characterizing their size, morphology and chemical composition. Silver and tellurium NS exhibited smaller size under anaerobiosis and their morphology was circular (silver NS), starred (tellurium NS) or amorphous (gold NS). Conclusions This nanostructure-synthesizing ability makes these isolates interesting candidates to get NS with biotechnological potential.

Background Kanamycin, an aminoglycoside, is an effective broad-spectrum antimicrobial agent against bacterial infections. However, the clinical efficacy of aminoglycosides has been overshadowed by the emergence of resistance mechanisms involving enzyme-mediated covalent modifications. Aminoglycoside nucleotidyltransferases (ANTs) inactivate aminoglycosides via AMP group transfer. Elucidating their molecular mechanisms and identifying effective inhibitors are critical for combating antimicrobial resistance. Methods Exiguobacterium sp . PL221A was isolated from hospital sewage under selection with 16 mg/L aminoglycosides. Following genomic sequencing using the Oxford Nanopore and BGISEQ-500 platforms, the eanT -1 gene encoding an ANT was identified, cloned, and expressed in Escherichia coli DH5α. Minimum inhibitory concentrations (MICs) against kanamycin were determined. Protein–ligand interactions between Eant-1 and kanamycin, virtual mutations of EanT-1, and inhibitor screening were assessed using the MaXFlow platform. To evaluate the catalytic mechanism, eant -1 mutants were expressed in vitro and their kinetic parameters were characterized. The impact of bicuculline on MICs and time-kill curves was also evaluated. Results Strain PL221A exhibited notable resistance to neomycin, kanamycin, and gentamicin (all MICs > 128 mg/L). Heterologous expression indicated that EanT-1 confers kanamycin resistance. Mutation of residues D43A and D98A abrogated enzymatic activity, whereas alanine substitutions at R51 and T100 had no apparent effect. Bicuculline, an isoquinoline alkaloid compound, was identified as an effective EanT-1 inhibitor, showing stronger binding affinity (˗8.72 kcal/mol) for EanT-1 than kanamycin (˗7.51 kcal/mol) and an ability to occupy aspartate residues critical for electrophilic polarization. Its hydroxyl-free alkaloid scaffold may prevent adenylation, enabling competitive binding with kanamycin at the catalytic site and inactivating EanT-1. Additionally, the combination of kanamycin and bicuculline effectively inhibited the growth of eanT -1–expressing bacteria. Conclusions The ANT encoded by eanT -1 in Exiguobacterium sp. PL221A mediates resistance to kanamycin, which can be counteracted by the alkaloid bicuculline through inhibition of its catalytic activity.

Litchi downy blight (LDB) caused by Peronophythora litchii destroys 20–30% of litchi fruit every year and causes significant economic losses. Some Exiguobacterium strains exhibit considerable promise in both agricultural and industrial sectors. E. acetylicum SI17, isolated from the litchi fruit carposphere, demonstrated significant biocontrol activity against LDB through pre-harvest treatment. To elucidate its underlying regulatory mechanisms, the genome of SI17 was sequenced and analyzed, revealing a circular chromosome spanning 3,157,929 bp and containing 3541 protein-coding genes and 101 RNA genes. Notably, 94 genes were implicated in the production of secondary metabolites. Among the 29 Exiguobacterium strains so far sequenced, SI17 possessed the largest genome. In the phylogenomic analysis encompassing the entire genome, SI17 was clustered into Group I. Treating litchi fruit with SI17 before harvesting resulted in a decrease in H2O2 content in the fruit peel and an increase in superoxide dismutase activity, thus enhancing resistance to LDB. Interestingly, SI17 did not display plate antagonism against Peronophythora litchii SC18. It can be inferred that SI17 generates secondary metabolites, which enhance litchi’s resistance to LDB. This study represents the first documentation of an Exiguobacterium strain exhibiting a role in litchi plant disease and showcasing significant potential for the biological control of LDB.

Colorectal cancer is the most common type of gastrointestinal cancers with poor survival and limited therapeutic options. In this study, four structurally different cyclic dipeptides (or diketopiperazine) were isolated and identified as cyclo ( l -Pro- l -Leu), cyclo ( l -Pro- l -Val), cyclo ( l -Pro- l -Phe) and cyclo ( l -Pro- l -Tyr) from the ethyl acetate extract in the cell-free filtrate of Exiguobacterium acetylicum S01. The anticancer potential of identified DKPs on colorectal cancer HT-29 cells in vitro and in vivo zebrafish xenograft model was evaluated. The MTT (3-(4, 5-dimethylthiazol-2yl)-2, 5-diphenyltetrazolium bromide)) assay showed that four DKPs exhibited significant inhibition of HT-29 cells viability in a dose-dependent manner whereas there were no cytotoxic effects on normal mouse fibroblast 3T3 cells. Also, we observed that all DKPs induce early and late apoptotic cell death in HT-29 cells. Moreover, the expression levels of apoptotic (cytochrome-c, caspase-3 and Bid) and anti-apoptotic (Bcl-2) markers were up- and down-regulated in HT-29 cells in response to DKPs treatments. Furthermore, these four DKPs remarkably inhibited the tumor progression in a zebrafish xenograft model within a nonlethal dose range. Overall, our findings suggest that cyclic dipeptides derived from E. acetylicum S01 could be promising chemopreventive/ therapeutic candidates against cancer.

The potential of antagonistic bacteria isolated from tomato rhizosphere soils of Oman in the control of damping-off disease of tomato was investigated. A total of 27 bacterial isolates were isolated from 18 soil samples collected from the rhizosphere of tomato from Al-Batinah South, Al-Sharqia North and Muscat Governorate. These bacterial isolates were tested in vitro for their antagonistic activity against Pythium aphanidermatum using a dual culture technique. Of the 27 bacterial isolates tested, four isolates designated D1/3, D1/8, D1/17 and D1/18 were effective in inhibiting the mycelial growth of P. aphanidermatum, by inducing an inhibition zone of 32.3, 10.3, 6.3 and 9.9 mm, respectively. Compatibility tests using a cross-streak assay on nutrient agar medium indicated that these four bacterial isolates were compatible with one another. The bacterial isolates were identified as Klebsiella oxytoca (D1/3), Exiguobacterium indicum (D1/8) and Bacillus cereus (D1/17 and D1/18), on the basis of the rRNA gene sequences. Among the isolates tested for in vitro plant growth promoting activity, D1/8 induced the maximum shoot length and seedling vigor. The potential of bacterial antagonists either individually or in combination in the control of damping-off disease of tomato was tested under greenhouse conditions. Among the biocontrol treatments, the combined application of D1/8 and D1/17 was the most effective, where damping-off incidence was reduced by 27 % relative to the infected control. These bacterial antagonists appear to be potential candidates to be developed as bio-inoculants for the ecofriendly management of damping-off of tomato under desert farming ecosystem.

The study of arsenic (As)-resistant microorganisms with high As removal capacity is fundamental for the development of economically sustainable technologies used for the treatment of water contaminated with metalloid. In the current study, four bacterial strains were isolated from As-contaminated water samples of the Xichu region, Mexico. Based on 16S rRNA gene sequencing and phylogenetic analysis of the isolated strains, Rhodococcus gordoniae, Microbacterium hydrocarbonoxydans, Exiguobacterium indicum, and Pseudomonas kribbensis were identified as potential As removal strains. R. gordoniae shows the highest growth capacity in both As(III) and As(V). R. gordoniae, M. hydrocarbonoxydans, and E. indicum removed approximately 81.6, 79.9, and 61.7% of As(III), as well as 77.2, 68.9, and 74.8% of As(V), respectively. P. kribbensis removed only about 80.2% of As(V). This study contributes to the possible detoxification mechanisms employed by these bacteria. Such insight could be crucial in the successful implementation of in situ bioremediation programs using these little-known bacteria.