Bacterium Exiguobacterium Research Articles

Constructed wetland coupled microbial fuel cell (CW-MFC) with Phragmites australis planted for hexavalent chromium removal and electricity generation

Hexavalent chromium (Cr(VI)) in aqueous solution is characterized by strong mobility, high solubility, and significant biological toxicity. In this study, four constructed wetland combined with microbial fuel cell (CW-MFC) systems were established to explore the changes in electricity generation, water purification, and microbial community structure under the influence of plant, bioelectrochemical, and Cr(VI) stress. The planted CW-MFC (total Cr: 97.5 %, Cr(VI): 99.2 %) and non-planted CW-MFC (total Cr: 95.3 %, Cr(VI): 97.3 %) showed significantly higher Cr removal efficiency than open circuit CW-MFC (total Cr: 86.9 %, Cr(VI): 84.8 %). The presence of plants and Cr(VI) increased the average voltage and power density of CW-MFC. Additionally, plants, bioelectrochemistry, and Cr(VI) improved COD removal and significantly enhanced the abundance and diversity of microorganisms in the system during long-term domestication. The Cr(VI)-reducing bacterium Exiguobacterium was least abundant in non-Cr(VI) containing CW-MFC system. The abundance of the electrochemically active bacteria (EAB) Geobacter increased under bioelectrochemistry and Cr(VI).

Marine microbial exopolysaccharide route synthesis of Ag and Pd metal nanoparticles: A possible anticancer, and antioxidant applications

The present research aimed develop an innovative and efficient method for fabricating silver and palladium nanoparticles (NPs)by using exopolysaccharide (EPS) from the seaweed-associated endo-symbiotic bacteria Exiguobacterium profundum (OP919469). Through, the application is a combined precipitation approach, EPS acts as a reducing and stabilising agent for the production of NPs. The major functional groups of NPs such as C–H, N–H, C–C, and C–O stretching’s were detected. The structural morphology of NPs showed a mixture of needle, spherical, and triangle-shaped with size between 10 and 50 nm. The purity of EPS was confirmed by high-performance liquid chromatography which revealed 99.81% purity. The NPs revealed strong anti-cancer and antioxidant properties. In addition, the NPs have been shown to dose-dependent toxicity with reduced hatching, survival rate, and abnormalities in zebrafish. The biochemical assay revealed that the EPS loaded NPsdid not induce oxidative stress up to 120 hpf in contrast to hydrogen peroxide which had a significant effect (P < 0.0001) on glutathione-S-transferase, catalase (P < 0.0017), and superoxide dismutase (P < 0.0001). Overall, our finding indicates that the molecular and functional characteristics of EPS and EPS loaded NPs represent future potential applications in the drug delivery and healthcare sectors.

A novel marine bacterium Exiguobacterium marinum a-1 isolated from in situ plastisphere for degradation of additive-free polypropylene

As the ecological niche most closely associated with polymers, microorganisms in the ‘plastisphere’ have great potential for plastics degradation. Microorganisms isolated from the ‘plastisphere’ could colonize and degrade commercial plastics containing different additives, but the observed weight loss and surface changes were most likely caused by releasing the additives rather than actual degradation of the plastics itself. Unlike commercial plastics that contain additives, whether marine microorganisms in the ‘plastisphere’ have adapted to additive-free plastics as a surface to colonize and potentially degrade is not yet known. Herein, a novel marine bacterium, Exiguobacterium marinum a-1, was successfully isolated from mature ‘plastisphere’ that had been deployed in situ for up to 20 months. Strain a-1 could use additive-free polypropylene (PP) films as its primary energy and carbon source. After strain a-1 was incubated with additive-free PP films for 80 days, the weight of films decreased by 9.2%. The ability of strain a-1 to rapidly form biofilms and effectively colonize the surface of additive-free PP films was confirmed by Scanning Electron Microscopy (SEM), as reflected by the increase in roughness and visible craters on the surface of additive-free PP films. Additionally, the functional groups of –CO, -C-H, and –OH were identified on the treated additive-free PP films according to Fourier Transform Infrared (FTIR). Genomic data from strain a-1 revealed a suite of key genes involved in biosurfactant synthesis, flagellar assembly, and cellular chemotaxis, contributing to its rapid biofilm formation on hydrophobic polymer surfaces. In particular, key enzymes that may be responsible for the degradation of additive-free PP films, such as glutathione peroxidase, cytochrome p450 and esterase were also recognized. This study highlights the potential of microorganisms present in the ‘plastisphere’ to metabolize plastic polymers and points to the intrinsic importance of the new strain a-1 in the mitigation of plastic pollution.

Symbiotic hemolymph bacteria reduce hexavalent chromium to protect the host from chromium toxicity in Procambarus clarkii

Hexavalent chromium (Cr(VI)) is a cytotoxic heavy metal pollutant that adversely affects all life forms. Interestingly, the crustacean Procambarus clarkii exhibits a relatively high tolerance to heavy metals. The underlying mechanisms remain unclear. In this study, we investigated the role of symbiotic bacteria in P. clarkii in alleviating Cr(VI)-induced damage and explored their potential mechanisms of action. Through transcriptomic analysis, we observed that Cr(VI) activated P. clarkii's antimicrobial immune responses and altered the bacterial composition in the hemolymph. After antibiotic treatment to reduce bacterial populations, Cr(VI)-induced intestinal and liver damage worsened, and crayfish exhibited lower levels of GSH/CAT/SOD activity. The Exiguobacterium, the symbiotic bacteria in the hemolymph of P. clarkii, were proved to be primary contributor to Cr(VI) tolerance. Further investigation suggested that it resists Cr(VI) through the activation of the ABC transporter system and the reduction of Cr(VI) via the reductase gene nfsA. To validate the role of Exiguobacterium in Cr(VI) tolerance, crayfish treated with antibiotics then supplemented with Exiguobacterium H6 and recombinant E. coli (with the nfsA gene), reduced Cr(VI)-induced ovarian damage. Overall, this study revealed that the symbiotic bacteria Exiguobacterium can absorb and reduce hexavalent chromium, mitigating Cr(VI)-induced damage in P. clarkii. These findings provide new insights into hexavalent chromium tolerance mechanisms in crustaceans.

Structural and Biochemical Insights into Bis(2-hydroxyethyl) Terephthalate Degrading Carboxylesterase Isolated from Psychrotrophic Bacterium Exiguobacterium antarcticum.

This study aimed to elucidate the crystal structure and biochemically characterize the carboxylesterase EaEst2, a thermotolerant biocatalyst derived from Exiguobacterium antarcticum, a psychrotrophic bacterium. Sequence and phylogenetic analyses showed that EaEst2 belongs to the Family XIII group of carboxylesterases. EaEst2 has a broad range of substrate specificities for short-chain p-nitrophenyl (pNP) esters, 1-naphthyl acetate (1-NA), and 1-naphthyl butyrate (1-NB). Its optimal pH is 7.0, losing its enzymatic activity at temperatures above 50 °C. EaEst2 showed degradation activity toward bis(2-hydroxyethyl) terephthalate (BHET), a polyethylene terephthalate degradation intermediate. We determined the crystal structure of EaEst2 at a 1.74 Å resolution in the ligand-free form to investigate BHET degradation at a molecular level. Finally, the biochemical stability and immobilization of a crosslinked enzyme aggregate (CLEA) were assessed to examine its potential for industrial application. Overall, the structural and biochemical characterization of EaEst2 demonstrates its industrial potency as a biocatalyst.

Display of oligo-α-1,6-glycosidase from &lt;i&gt;Exiguobacterium sibiricum&lt;/i&gt; at the surface of &lt;i&gt;Escherichia coli &lt;/i&gt;cells

Cell-surface display based on the use of anchor motifs of outer membrane proteins allows the exposure of target peptides and proteins on the surface of microbial cells. Previously, we obtained and characterized recombinant oligo-α-1,6-glycosidase of the psychrotrophic bacterium Exiguobacterium sibiricum (EsOgl) which demonstrated high catalytic activity. It has also been shown that the autotransporter AT877 from Psychrobacter cryohalolentis and its deletion variants effectively exhibit the 10th domain of type III fibronectin (10Fn3) on the surface of Escherichia coli cells. The aim of the work was to obtain an EsOgl display system on the surface of bacterial cells based on AT877. The genes of the hybrid autotransporter EsOgl877 and its deletion mutants EsOgl877Δ239 and EsOgl877Δ310 were constructed. The enzymatic activity of EsOgl877 was investigated and it was found that the cells expressing this protein retained about 90% of the maximum activity in the range of 15-35°C. It was shown that activity of the cells containing EsOgl877Δ239 and EsOgl877Δ310 was 2.7 and 2.4 times higher, respectively, than of the cells expressing full-sized AT. Analysis of cells expressing shortened variants of EsOgl877 after treatment with proteinase K showed that the passenger domain is also localized on the cell surface. The obtained results can be used for optimization of the display systems of oligo-α-1,6-glycosidase and other heterologous proteins on the surface of E. coli cells.

Macroalgae-associated halotolerant marine bacteria Exiguobacterium aestuarii ADCG SIST3 synthesized gold nanoparticles and its anticancer activity in breast cancer cell line (MCF-7)

Marine microorganisms associated with macroalgae have currently been identified as an environmentally safe and efficient technique to use as biofactories in order to produce metal nanoparticles. The current investigation focuses to identify the capability of marine bacteria Exiguobacterium aestuarii ADCG SIST 3 for the extracellular synthesis of gold nanoparticles (AuNPs). The AuNPs that are synthesized were characterized by different techniques and analyzed for their applications in anticancer activity on normal and cancer cells. To the best of our knowledge, this study is first to systematically examine extracellular synthesis of AuNPs by the halotolerant bacteria Exiguobacterium aestuarii ADCG SIST 3 associated with seaweed. The characterization of synthesized AuNPs were carried out using Fourier Transform- Infrared Spectroscopy (FTIR), Raman Spectroscopy, Field Emission Scanning Electron Microscope (FE-SEM),High-Resolution Transmission Electron Microscope (HR-TEM), Particle size analyzer (DLS), Zetasizer, and UV–vis Spectrophotometer. With an average size of 37 nm, the fabricated AuNPs were observed to have a spherical-likemorphology. The cytotoxic response of synthesized AuNPs was assessed by MTT assay on Vero and MCF-7 cell lines. The antioxidant potential of the synthesized AuNPs and apoptosis studies were also carried out which resulted in excellent activity in the MCF-7 cells with potential application in the medical field.

Characterization of chitinase from Exiguobacterium antarcticum and its bioconversion of crayfish shell into chitin oligosaccharides

Crayfish have become a heavily consumed food and its chitin-rich shell is of great value in terms of waste conversion. This study found a novel chitinase (EaChi40) from a marine bacterium Exiguobacterium antarcticum. The gene was cloned and expressed as a soluble protein of 40 kDa, having optimal activity at pH 6.0 and 30 °C. EaChi40 showed good stability and high specific activity, and kinetic studies found Km and Vmax were 0.86 mg/mL and 13.66 μmol/min/mg. For conversion crayfish shell into oligosaccharides, ball milling and ultrasound-assisted hydrogen peroxide decolorization were applied to pretreat crayfish shell to facilitate its hydrolysis. After the enzymatic conversion, the hydrolysis products of chitobiose and N-acetyl-D-glucosamine were 9.09 mg/mL and 9.21 mg/mL, respectively. EaChi40 efficiently degraded crayfish with a high hydrolysis rate of 76.1%. It is expected to be a good candidate for the production of chitin oligosaccharides in the food and biological fields.

Assessment and characterization of poly(3-hydroxybutyrate) accumulated by endophytic bacterium Exiguobacterium acetylicum BNL 103 from oleaginous plant Brassica napus L.

Assessment and characterization of poly(3-hydroxybutyrate) accumulated by endophytic bacterium Exiguobacterium acetylicum BNL 103 from oleaginous plant Brassica napus L.

Multiple factors influence bacterial community diversity and composition in soils with rare earth element and heavy metal co-contamination

The effects of long-term rare earth element (REE) and heavy metal (HM) contamination on soil bacterial communities remains poorly understood. In this study, soil samples co-contaminated with REEs and HMs were collected from a rare-earth tailing dam. The bacterial community composition and diversity were analyzed through Illumina high-throughput sequencing with 16S rRNA gene amplicons. Bacterial community richness and diversity were lower in the co-contaminated soils than in the uncontaminated soils, with clearly different bacterial community compositions. The results showed that total organic carbon and available potassium were the most important factors affecting bacterial community richness and diversity, followed by the REE and HM contents. Although the canonical correspondence analysis results showed that an REE alone had no obvious effects on bacterial community structures, we found that the combined effects of soil physicochemical properties and REE and HM contents regulated bacterial community structure and composition. The effects of REEs and HMs on bacterial communities were similar, whereas their combined contributions were greater than the individual effects of REEs or HMs. Some bacterial taxa were worth noting. These specifically included the plant growth-promoting bacteria Exiguobacterium (sensitive to REEs and HMs) and oligotrophic microorganisms with metal tolerance (prevalent in contaminated soil); moreover, relative abundance of JTB255-Marine Benthic Group, Rhodobacteraceae, Erythrobacter, and Truepera may be correlated with REEs. This study was the first to investigate the responses of bacterial communities to REE and HM co-contamination. The current results have major implications for the ecological risk assessment of environments co-contaminated with REEs and HMs.

Engineering of Thermal Stability in a Cold-Active Oligo-1,6-Glucosidase from Exiguobacterium sibiricum with Unusual Amino Acid Content.

A gene coding for a novel putative amylase, oligo-1,6-glucosidase from a psychrotrophic bacterium Exiguobacterium sibiricum from Siberian permafrost soil was cloned and expressed in Escherichia coli. The amino acid sequence of the predicted protein EsOgl and its 3D model displayed several features characteristic for the cold-active enzymes while possessing an unusually high number of proline residues in the loops—a typical feature of thermophilic enzymes. The activity of the purified recombinant protein was tested with p-nitrophenyl α-D-glucopyranoside as a substrate. The enzyme displayed a plateau-shaped temperature-activity profile with the optimum at 25 °C and a pronounced activity at low temperatures (50% of maximum activity at 5 °C). To improve the thermal stability at temperatures above 40 °C, we have introduced proline residues into four positions of EsOgl by site-directed mutagenesis according to “the proline rule”. Two of the mutants, S130P and A109P demonstrated a three- and two-fold increased half-life at 45 °C. Moreover, S130P mutation led to a 60% increase in the catalytic rate constant. Combining the mutations resulted in a further increase in stability transforming the temperature-activity profile to a typical mesophilic pattern. In the most thermostable variant A109P/S130P/E176P, the half-life at 45 °C was increased from 11 min (wild-type) to 129 min.

Characterisation of Pigment Producing Bacteria Isolated from Bio Floc Ponds and Its Colour Enhancement Effect in Xiphophorus helleri

Background: Biofloc technology is a minimal or zero water exchange technology which exerts beneficial effects on water quality, improves feed conversion ratio by recycling microbial protein in the culture of commercially important finfishes and shellfishes. This culture technique also improves the pigmentation in the ornamental fish culture. The presence of pigment producing bacteria and the absorption of the pigments by the ornamental fishes under biofloc technology would pave a long way to boost the production and export market of the high value fishes. Therefore, the present study evaluated the use of bacterial pigments isolated from the biofloc culture in the diets of Swordtail ornamental fish for its colouration and pigment absorption ability.Methods: A 30 days trial was conducted to evaluate the colour enhancement in swordtail through the supplementation of pigment produced from the bacteria in their diets. Swordtail fishes (1.2 ± 0.01g) were stocked in 100 litres tanks (20nos / tank) in triplicates. Fish fed with normal diet served as control and diet supplemented with pigment (50mg/kg) isolated from the bacteria Exiguobacterium profundum (T1), Chryseobacterium joostei (T2), Staphylococcus pasteuri (T3), Staphylococcus arlettae (T4) served as treatments. The characteristic features of the pigments isolated from the four different bacteria were checked and showed enhanced antibacterial, total antioxidant activity and the reductive ability.Result: Significant difference was observed in water quality parameters except temperature between the experimental groups. Growth parameters showed significant difference between control and treatment. Highest carotenoid concentration was found in fishes fed with pigments extracted from Staphylococcus pasteuri compared to other experimental groups. The present study proved the incorporation of bacterial pigments in the diets of Swordtail enhanced the total carotenoid concentration.

Exiguobacterium aurantiacum virulent pigment producing a novel pathogenic bacteria associated with cases of corneal ulcers

Blindness is a major public health problem in most developing countries. Corneal ulcer (Keratitis) is a major cause of blindness throughout the world. About 10% cases of blindness are due to corneal ulcer. “Corneal ulcer means loss of corneal substances as a result of infection and formation of raw, excavated area.” Bacterial keratitis is an acute or chronic, transient or recurrent infection of the cornea with varying predilection for anatomical and topographical parts of the cornea like marginal or central. This study demonstrates the Novel pathogenic bacteria Exiguobacterium aurantiacum with bad pigment production, which are causing corneal ulcer and determine the best possible empirical therapy and specific therapy. Particular attention should be given to this condition as it can progress very rapidly with complete corneal destruction can be occur within 24–48 hours. Keywords: Corneal Ulcer, Exiguobacterium aurantiacum, Molecular characterization, Antibiotic treatment, Pigment.

A rationally identified marine GH1 β‐glucosidase has distinguishing functional features for simultaneous saccharification and fermentation

AbstractThe classical route for second‐generation ethanol from lignocellulosic biomass is hampered by high process costs, fostering the development of alternative strategies such as simultaneous saccharification and fermentation (SSF). However, the lack of compatible enzyme cocktails poses a challenge. In this study, the enzyme EmBgl from the marine bacterium Exiguobacterium marinum was rationally identified based on structural and phylogenetic analyses, known desirable properties of close orthologs, and the ecological niche of its organism source. EmBgl is a multifunctional and glucose‐tolerant enzyme that efficiently hydrolyzes cello‐oligosaccharides due to a positive‐subsite region that can accommodate long cello‐oligosaccharides without imposing steric impediments. The efficacy of EmBgl in an SSF process was demonstrated using pretreated sugarcane bagasse as feedstock, yielding 28 g L−1 of ethanol in 30 h. The distinguishing functional properties of EmBgl and its successful utilization in an SSF process highlight its potential in biotechnological applications in which lignocellulose deconstruction is desirable under milder temperatures © 2020 Society of Chemical Industry and John Wiley &amp; Sons, Ltd.

Bioconversion of chitin waste using a cold-adapted chitinase to produce chitin oligosaccharides

Chitin waste, a highly insoluble biopolymer from sea food industry, can be converted into value-added products with chitinase (EC 3.2.1.14). Here, we purified a cold-adapted chitinase (EaChi39) from the marine bacterium Exiguobacterium antarcticum DW2. The enzyme was mostly activated at 30 °C and remained 35% of the maximal activity at 0 °C. It also showed a broad range of pH stability, high thermostability but low Km values, indicating the strong ability for chitin degradation. As an endo-type chitinase, EaChi39 completely converted colloidal chitin into N-acetylglucosamine (GlcNAc) and oligosaccharides with high yields, which was determined by HPLC (GlcNAc 9.9 mg mL−1, GlcNAc2 14.8 mg mL−1 and GlcNAc3 5.0 mg mL−1). As a novel chitinase from Exiguobacterium genus, EaChi39 displayed advantageous cold-adaptation and thermostability. It may be a superior candidate for bioconversion of seafood by-products in producing bioactive oligosaccharides.

Characterization of Struvite Produced by an Algal Associated Agarolytic Bacterium Exiguobacterium aestuarii St. SR 101

Characterization of Struvite Produced by an Algal Associated Agarolytic Bacterium Exiguobacterium aestuarii St. SR 101

Production, optimization and partial purification of protease from terrestrial bacterium Exiguobacterium profundam sp. MM1

Abstract A new potent protease producing terrestrial bacterium was isolated and identified as Exiguobacterium profundam sp. MMI by 16 S rRNA gene sequencing. Protease production was enhanced by optimizing the culture conditions. The nutritional factors such as carbon and nitrogen sources, substrate sources and also physical parameters like incubation time, pH, and temperature were optimized for the maximum yield of protease. Studies on the effect of different carbon and nitrogen sources revealed that glucose and peptone enhances the enzyme production. The optimum temperature and pH for enzyme production were at 35 °C and pH 9 respectively. The protease enzyme from Exiguobacterium profundam sp. MMI MG951843.1 was partially purified by ammonium sulphate precipitation and was electrophoresed on 10% (w/v) SDS-PAGE using standard protein markers. The electrophoretic profile of the partially purified enzyme extract shows seven bands with the value of PM obtained from the curve calibration markers to known weight were 48.3KDa. 37.2 kDa, 28.2 kDa, 26.5 kDa, 24.5 kDa,20.4 kDa,16.5 kDa. The band with 48.3KDa shows presence of protease in the sample and further the caseinolytic activity of the partially purified protease enzyme was determined by zymogram.

Impact of mild alkali dosage on immobilized Exiguobacterium spp. mediated cost and energy efficient sludge disintegration

Approaches to (extracellular polymeric substance) EPS removal were studied with major aim to enhance the biodegradability and sludge solubilization. In this study, a novel approach of entrapment of bacterial strain was carried out to achieve long term activity of protease secreting bacteria Exiguobacterium sp. A mild treatment of potassium hydroxide (KOH) was applied to remove EPS which was followed by entrapment under the biological pretreatment. The efficiency of Exiguobacterium was predicted through dissolvable organic and suspended solids (SS) reduction. The maximum dissolvable organic matter released was 2300mg/L with the solubilization of 23% which was obtained for sludge without EPS (SWOE). For dissolvable organic release, SWOE showed higher final methane production of 232mL/g COD at the production rate of 16.2mL/g COD.d. The SWOE pretreatment was found to be cost effective and less energy intensive beneficial in terms of energy and cost (43.9KWh and −8.2USD) when compared to sludge with EPS (SWE) pretreatment (−177.6KWh and −91.23USD).

Characterization of an L-phosphinothricin resistant glutamine synthetase from Exiguobacterium sp. and its improvement.

A glutamine synthetase (GS; 1341bp) gene with potent L-phosphinothricin (PPT) resistance was isolated and characterized from a marine bacterium Exiguobacterium sp. Molecular docking analysis indicated that the substitution of residues Glu60 and Arg64 may lead to significant changes in binding pocket. To enhance the enzymatic property of GS, variants E60A and R64G were obtained by site-directed mutagenesis. The results revealed a noteworthy change in the thermostability and activity in comparison to the wild type (WT). WT exhibited optimum activity at 35°C, while E60A and R64G exhibited optimum activity at 45 and 40°C, respectively. The mutant R64G was 4.3 times more stable at 70°C in comparison to WT, while E60A was 5.7 times more stable. Kinetic analysis revealed that the k cat value of R64G mutant was 8.10-, 7.25- and 7.63-fold that of WT for ADP, glutamine and hydroxylamine, respectively. The kinetic inhibition (K i, 4.91±0.42mM) of R64G was 2.02-fold that of WT (2.43±0.14mM) for L-phosphinothricin. The analysis of structure and function relationship showed that the binding pocket underwent dramatic changes when Arg site of 64 was substituted by Gly, thus promoting the rapid capture of substrates and leading to increase in activity and PPT-resistance of mutant R64G. The rearrangements of the residues at the molecular level formed new hydrogen bonds around the active site, which contributed to the increase of thermostability of enzymes. This study provides new insights into substrate binding mechanism of glutamine synthetase and the improved GS gene also has a potential for application in transgenic crops with L-phosphinothricin tolerance.

Crystal Structure and Functional Characterization of an Esterase (EaEST) from Exiguobacterium antarcticum.

A novel microbial esterase, EaEST, from a psychrophilic bacterium Exiguobacterium antarcticum B7, was identified and characterized. To our knowledge, this is the first report describing structural analysis and biochemical characterization of an esterase isolated from the genus Exiguobacterium. Crystal structure of EaEST, determined at a resolution of 1.9 Å, showed that the enzyme has a canonical α/β hydrolase fold with an α-helical cap domain and a catalytic triad consisting of Ser96, Asp220, and His248. Interestingly, the active site of the structure of EaEST is occupied by a peracetate molecule, which is the product of perhydrolysis of acetate. This result suggests that EaEST may have perhydrolase activity. The activity assay showed that EaEST has significant perhydrolase and esterase activity with respect to short-chain p-nitrophenyl esters (≤C8), naphthyl derivatives, phenyl acetate, and glyceryl tributyrate. However, the S96A single mutant had low esterase and perhydrolase activity. Moreover, the L27A mutant showed low levels of protein expression and solubility as well as preference for different substrates. On conducting an enantioselectivity analysis using R- and S-methyl-3-hydroxy-2-methylpropionate, a preference for R-enantiomers was observed. Surprisingly, immobilized EaEST was found to not only retain 200% of its initial activity after incubation for 1 h at 80°C, but also retained more than 60% of its initial activity after 20 cycles of reutilization. This research will serve as basis for future engineering of this esterase for biotechnological and industrial applications.