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. 2026 Jul 3;17:1799206. doi: 10.3389/fphar.2026.1799206

Experimental analysis and genome mining for functional validation of genes associated with anti-inflammatory, antioxidant, and antibacterial activities in Kurthia gibsonii VITAM20

Anju Kanjirakandi Ashokan 1, Mohanasrinivasan Vaithilingam 1,*,†
PMCID: PMC13375520  PMID: 42490912

Abstract

Introduction

The identification of microbial strains harbouring bioactive genes is important for the discovery of novel therapeutic agents. The study objective was to identify the genes in Kurthia gibsonii VITAM20 that are associated with anti-inflammatory and antioxidant properties and validate these findings with in vitro functional assays.

Methods

Genes associated with anti-inflammatory and antioxidant activities were identified by BLAST analysis against curated databases. The anti-inflammatory, antioxidant, and antibacterial potential of Kurthia gibsonii VITAM20 were subsequently evaluated using in vitro assays.

Results

BLAST analysis identified several genes associated with anti‐inflammatory properties, including eps (95.455%), NEMO (100%), spa (100%), and spaB (90.625%). Multiple genes associated with antioxidant potential were also detected, including CAT (92.308%), dps (95.238%), gshR3 (91.304%), gshR4 (100%), KATA (100%), oxyR (100%), KatA (88.462%), perR (74.041%, bit score 126), sodA (100%), sodM (76.471%), and trxA (77.778%). The genomic findings were supported by in vitro analyses, which demonstrated notable anti‐inflammatory activity. Kurthia gibsonii VITAM20 exhibited human red blood cell membrane stabilization rates of 90.78% ± 1.03% and 69.92% under non-heated and heated conditions, respectively, compared with diclofenac (83.01% ± 0.72% and 83.83% ± 1.23%). In the egg albumin denaturation assay, Kuthia gibsonii VITAM20 demonstrated a concentration‐dependent increase in inhibition from 80.94% ± 0.57% to 84.47% ± 0.39%, comparable to that of diclofenac. Furthermore, Kurthia gibsonii VITAM20 exhibited 64.27% ± 0.46% antioxidant activity compared with ascorbic acid (91.40% ± 0.46%), with an IC50 value of 286.5 µg/mL. The crude enzyme extract demonstrated antibacterial activity against Listeria monocytogenes and Staphylococcus aureus, producing inhibition zones of 13 mm. The MIC was determined to be 250 µg/mL for both pathogens. At this concentration, the crude enzyme extract inhibited the growth of L. monocytogenes and S. aureus by 81.46% and 77.82%, respectively.

Discussion

The combined genomic and functional analyses confirmed the presence of conserved genes associated with anti‐inflammatory and antioxidant activities in Kurthia gibsonii VITAM20. These findings suggest that Kurthia gibsonii VITAM20 represents a promising source of bioactive molecules with potential therapeutic applications.

Keywords: anti-inflammatory protease, bioactive molecule, inflammation, Kurthia gibsonii, secondary metabolites

1. Introduction

Inflammation is a key biological response to injury or stimuli, such as the entry of pathogens (Gusev and Zhuravleva, 2022). This process involves vasodilation, increased vascular permeability, and plasma leakage, which causes pain, elevated body temperature, and swelling (Melnikova et al., 2018). Traditional anti-inflammatory remedies have been widely used in the treatment of inflammatory conditions (Ali et al., 2023). However, these drugs usually cause side effects, including digestive problems and kidney failure, while their continued usage leads to the increase of reduced therapeutic effectiveness and drug resistance (Sohail et al., 2023). In response to the existing limitations, enzyme-based treatments have shifted toward varied approaches, such as high specificity, catalytic efficacy, and reduced adverse effects.

Enzymes produced by bacteria exhibit increased bioactivity, including anti-inflammatory, antioxidant, and antibacterial properties. These enzymes are effective against inflammation through regulating the inflammatory signaling pathways and inhibiting the production of pro-inflammatory agents (Liu et al., 2022). The antioxidant properties of these enzymes are involved in the absorption of reactive oxygen species (ROS) and the chelation of metal ions, which are significant for reducing oxidative stress (Tumilaar et al., 2024). In response to the existing limitations, enzyme-based treatments have shifted toward varied approaches, such as high specificity, catalytic efficacy, and reduced adverse effects.

Additionally, these bacterial enzymes demonstrated a notable antibacterial effect by degrading the walls of microbial cells and interrupting biofilm development, which consequently decreased the spread and strength of disease-causing microorganisms. The role of inflammation is significant in host defense and tissue repair, yet, when there is an imbalance, it can often lead to different acute and chronic diseases, encompassing autoimmune conditions, tumors, metabolic issues, and heart-related problems. The current study focused on a bacterium named Kurthia gibsonii VITAM20 isolated from butcher soil samples collected in Vellore, Tamil Nadu. The antioxidant, antimicrobial, and anti-inflammatory properties of this isolate were systematically evaluated by genome mining and confirmed by in vitro assays. This approach aimed to find novel bioactive enzymes with potential therapeutic and industrial applications.

2. Materials and methods

2.1. Isolation and identification of the bacteria

Soil samples were taken from Viruthampet (12.94152°N, 79.1351°E), Vellore, Tamil Nadu, India. Serial dilutions were performed, and aliquots were spread on agar plates. The plates were kept at 37 °C for 48 h. After incubation, plates showing different microbial colonies were chosen for further studies. A potent bacterial isolate named VITAM20 was identified through morphological, biochemical, and whole-genome sequencing (Chauhan and Samant, 2022). (The datasets generated and analyzed during the current study are available under the name Mohanasrinivasan V repository; Submission ID: SUB15186439, BioProject ID: PRJNA1238542, BioSample: SAMN47475936, SRA: SRR35894991, Organism: Kurthia gibsonii VITAM20; https://www.ncbi.nlm.nih.gov/sra/PRJNA1238542.)

2.2. Determination of the growth curve

A single bacterial colony was inoculated into fresh liquid nutrient broth medium and incubated overnight at 30 °C and 250 rpm until the culture reached a stationary phase. After that, 1 mL of the inoculum was transferred to 100 mL of nutrient broth and incubated at 30 °C and 250 rpm for 48 h, along with setting up a control. Optical density at 600 nm was measured at every 2-h intervals (Roy et al., 2025).

2.3. Identification of reference genes in the Kurthia gibsonii VITAM20 genome

For the identification of a few reference genes associated with anti-inflammatory and antioxidant activity, we implemented a customized analysis pipeline built on the BLAST+ suite and supported by automated parsing and filtering routines. Reference sequences of interest, consisting of eps, NEMO, dps, gshR3, gshR4, KatA, spa, spaB, CAT, oxyR, KatA_pseudo, perR, sodA, SodM, and trxA, were retrieved from NCBI and prepared in the FASTA format as query inputs (the query inputs are given as Supplementary Table S1). The assembled genome of Kurthia gibsonii VITAM20, containing nearly 2.9 million base pairs, was formatted into a searchable BLAST+ database using the “makeblastdb” utility. Query sequences were searched against this database, and alignments were retrieved in tabular format, which provided information on sequence identity, alignment length, mismatches, bit score, and statistical significance. Alignments meeting the defined quality thresholds, percentage identity ≥70%, and E-value ≤1e-5 were maintained for further analysis.

2.4. HRBC membrane stabilization method

Blood samples were drawn from healthy individuals and mixed with an equivalent volume of Alsever's solution (2% dextrose, 0.8% sodium citrate, 0.05% citric acid, and 0.42% sodium chloride in distilled water). For the collection of human blood, an approval from the institutional ethical committee for human studies (VIT/IECH/2025/16 IECH/15 February 2025/19) was obtained. The sample was then centrifuged for 10 min at 3,000 rpm, and concentrated red blood cells were cleaned using isotonic saline with 0.9% NaCl. A 10% suspension of packed red blood cells (HRBC) was then formulated using isotonic saline. Different volumes of the crude samples were prepared by utilizing distilled water, which involved mixing phosphate buffer (1 mL), hypotonic saline (2 mL), and HRBC solution (0.5 mL). The assay mixtures were kept constant for 30 min at 37 °C and then spun at 3,000 rpm for 20 min to collect the liquid on the top for further evaluations (Joseph et al., 2013). Diclofenac was used as the reference standard. All the experiments were carried out in triplicates (n = 3). The percentage of HRBC membrane protection was calculated using the following formula.

% Protection=100−OD of test sample÷OD of control×100.

The percentage of hemolysis of the HRBC membrane can be calculated using following equation.

% Hemolysis=OD of test sample÷OD of control×100.

2.5. Heat-induced hemolysis

The hemolysis experiment, induced by thermal conditions, evaluated the anti-inflammatory effects of the bacterial crude sample under temperature stress. Sample concentrations of 50, 100, 200, 400, and 800 μg/mL were prepared in ethanol using isotonic phosphate buffer. Tubes were incubated for 20 min at 54 °C in a regulated water bath. After centrifuging the mixture for 5 min at 5,000 rpm, the absorbance was quantified at 560 nm. Diclofenac was used as the reference standard (Akwayamai John and Henry Bulama, 2020).

The following formula was used to calculate the percentage inhibition of hemolysis.

% Protection=100− OD of test sample÷OD of control×100.

2.6. Egg albumin denaturation assay

The anti-inflammatory efficiency of the crude extract sample was evaluated using the denaturation of albumin assay, following the method described by Hasan et al. (2024) with minor modifications. The reaction mixture consisted of 1 mL of phosphate buffer (pH 6.5), 200 μL of hen’s egg albumin, and various concentrations from hen’s egg albumin, and various concentrations of the crude sample. An equal volume of distilled water served as the control. The reaction mixtures then heated and incubated at 37 °C for 15 min and then later received a thermal treatment at 70 °C for a span of 5 min. The absorbance was taken after the cooling process at 660 nm. All the experiments were carried out in triplicates (n = 3). Diclofenac was used as a reference standard drug (Chaiya et al., 2022). The percentage inhibition was calculated in accordance with the following formula.

% Inhibition=1− Absorbance of sample÷Absorbance of control×100.

2.7. Antioxidant activity assay

The antioxidant potential of the bacterial isolate was analyzed using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical-scavenging activity. Different volumes of the supernatant obtained from the freshly inoculated culture were mixed with DPPH (2.5 mL) solution, and the total volume was adjusted to 3 mL with the addition of distilled water. The mixture was agitated thoroughly and incubated for 30 min at 37 °C in the dark. The absorbance was later measured at 515 nm. Ascorbic acid was used as the standard reference (Pourramezan et al., 2020). The percentage of free radical scavenging activity was calculated using the following equation.

% Inhibition= Absorbance of control−Absorbance of sample÷Absorbance of control×100.

2.8. Statistical analysis

All experiments were performed with clearly defined biological triplicates. GraphPad Prism software version 9.0.0 (GraphPad Software, San Diego, CA, United States) was used for statistical analysis for the determination of the difference among samples. The results were expressed as the mean ± standard deviation (SD). Two-way ANOVA using Sidak’s multiple comparisons test was performed.

2.9. Antibacterial activity

Kurthia gibsonii VITAM20’s antibacterial activity was analyzed against Gram-positive bacteria, especially Staphylococcus aureus (MTCC 3160) and Listeria monocytogenes (MTCC 657), along with its effects on Gram-negative bacteria including Escherichia coli (MTCC 443) and Pseudomonas aeruginosa (MTCC 2582). This experiment utilized ciprofloxacin as the standard reference. The antibacterial activity was analyzed according to the recommendations established by the National Committee for Clinical Laboratory Standards (NCCLS) utilizing the agar well diffusion procedure. The bacteria were grown in the nutrient broth for a duration of 24 h (1.5 × 10^8 CFU/mL) and swabbed onto sterile Mueller–Hinton agar plates. Following that, a sterile well punch was used to punch wells with a diameter of 6 mm, and the crude sample supernatant (100 μg/mL) was added. The agar plates were later incubated at 37 °C for a full day, using unprocessed sterile nutrient broth as the negative control for comparison (Valgas et al., 2007). The minimum inhibitory concentration (MIC) of the crude enzyme extract was determined according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI), 2010 using the broth microdilution method in Mueller–Hinton broth. A bacterial inoculum (1.5 × 10^8 CFU/mL) was prepared from 24-hour-old cultures and standardized to the 0.5 McFarland standard. The crude extract was tested against Staphylococcus aureus and Listeria monocytogenes. Ciprofloxacin was used as the positive control at a concentration of 2 μg/mL, while distilled water served as the solvent control, and Mueller–Hinton broth served as the sterility control. In each well of a 96-well microplate, 100 µL of Mueller–Hinton broth, 10 µL of the standardized bacterial suspension, and 100 µL of the crude enzyme extract at concentrations ranging from 9.06 to 1000 μg/mL were added. The plates were then incubated at 37 °C for 24 h, and the optical density was measured at 600 nm. All experiments were performed in triplicate (n = 3). The MIC determination using the resazurin-based 96-well microtiter plate method is shown in the supplementary image (Al-Mijalli et al., 2023).

3. Results

3.1. Identification of VITAM20

The VITAM20 strain was identified as Kurthia gibsonii by whole-genome sequencing and characterization (Oxford Nanopore Technologies). The genomic DNA of the strain was successfully extracted and quantified. The NanoDrop analysis revealed a DNA concentration of 172.1 ng/μL, with A260/280 and A260/230 ratios of 1.86 and 2.29, respectively, indicating high purity of the extracted DNA. The GC content of the strain Kurthia gibsonii VITAM20 was 36.5%. Genome assembly was performed from scratch with Unicycler, and then scaffolding with Ragout led to the draft genome of approximately 2.97 Mb for VITAM20. BLAST-based identification results are provided in Supplementary Material to support the taxonomic placement of the strain.

3.2. Growth curve

The growth curve showed a typical microbial growth pattern with distinct phases. Initially, a lag phase was observed (0 h–4 h), during which a minimal increase in optical density indicated cellular adaptation to the new environment. This was followed by a log phase (6 h–28 h), characterized by a rapid and consistent increase in growth, which reflected active cell division and metabolic activity. The growth reached a maximum at approximately 34 h –36 h, indicating the onset of the stationary phase, where the rate of cell proliferation balanced the rate of cell death. Later, a gradual decline in growth was observed from 38 h onward, marking the death phase, during which a reduction in cell density indicated loss of viability and cell lysis. Overall, the strain exhibited a distinct growth profile with the peak biomass achieved at approximately 36 h (Figure 1).

FIGURE 1.

Line graph showing absorbance at 600 nanometers on the vertical axis and time in hours on the horizontal axis. Absorbance starts near zero, rises steadily to a peak around 36 hours, then gradually decreases.

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Growth curve of Kurthia gibsonii VITAM20.

3.3. Computational identification of functional gene clusters

The whole-genome characterization of the bacterial isolate revealed that it was a Kurthia gibsonii VITAM20 strain. To assess the presence of few reference genes responsible for the anti-inflammatory and antioxidant properties within the Kurthia sp. Genome, a nucleotide sequence resemblance analysis was conducted using a custom Python-based pipeline. The evaluation yielded high-confidence alignments for various target genes. The eps gene demonstrates 95.455% nucleotide identity, with an E-value of 0.037 and a bit score of 36.2. The NEMO gene also exhibits complete identity (100%) with an E-value of 0.39 and a bit score of 32.5. The p75 gene shares 95% identity, with an E-value of 0.7 and a bit score of 32.5. The spaA gene shows 100% sequence identity, confirmed by a low E-value of 0.033 and a bit score of 38.1. The spaB gene shows the lowest percentage identity (90.625%), albeit with a highly significant E-value of 0.000686 and the highest bit score among the identified genes (43.6). The CAT gene exhibits 92.308% nucleotide similarity, with an E-value of 0.015 and a bit score of 38.1. The dps gene determines 95.238% identity (E-value 0.065, bit score 34.4), while the gshR3 gene shares 91.304% identity (E-value 0.64, bit score 32.5). Complete sequence identity (100%) is observed for the gshR4 gene (E-value 0.63, bit score 32.5), the KatA gene (E-value 0.69, bit score 32.5), and the oxyR gene (E-value 0.033, bit score 36.2). The KatA gene shows 88.462% sequence identity (E-value 0.69, bit score 32.5). The perR gene, although presenting a lower percentage identity of 74.041%, exhibits a highly significant E-value of 8.94 × 10−30 with the highest bit score among this set (126), signifying strong alignment confidence despite divergence. The sodA gene also displays complete identity (100%) with an E-value of 0.28 and bit score of 32.5, while the sodM gene shares 76.471% identity (E-value 0.022, bit score 36.2). Finally, the trxA gene records 77.778% identity with an E-value of 0.000886 and a bit score of 39.9.

3.4. Detection of secondary metabolites using antiSMASH

Using the antiSMASH software designed for detecting secondary metabolites, we identified a biosynthetic pathway associated with terpenes and their precursors. In addition, we observed that there is no similarity confidence because these are categorized into unidentified clusters. These compounds contributed to the increased functionality of bacteriocins by providing additional antimicrobial metabolites. We also identified key genes that are essential to produce cyclic-lactone auto-inducers, which revealed a quorum-sensing network that influenced the activation of bacteriocin gene clusters (Figure 2).

FIGURE 2.

Three annotated gene cluster schematic diagrams for scaffold_1 show regions labeled as terpene or terpene-precursor, each with color-coded gene types including core biosynthetic, additional biosynthetic, transport-related, regulatory, other genes, resistance, and binding sites; legends and location coordinates are provided for each region.

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Secondary metabolite production pathways predicted by antiSMASH, including terpene-precursor and terpene.

3.5. Results for anti-inflammatory assays

In vitro studies of Kurthia gibsonii VITAM20 indicated that the isolate possesses remarkable anti-inflammatory properties against inflammation. The results are summarized in Figure 3. Compared to the stabilization reference outlined by diclofenac, the crude enzyme sample (1,000 μg/mL) revealed a notable stabilization of 90.78% ± 1.03%.

FIGURE 3.

Bar graph comparing percentage of protection in an HRBC membrane stabilization assay at five concentrations for VITAM20 and Diclofenac, with separate bars for stabilization and hemolysis; VITAM20 shows higher stabilization and lower hemolysis at all concentrations, with statistical significance indicated by asterisks.

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HRBC membrane stabilization assay showing the percentage protection at different concentrations of crude enzyme extract from Kurthia gibsonii VITAM20 and the standard drug diclofenac. All experiments were performed in triplicate (n = 3), and data are presented as the mean values.

For the different concentrations of samples, Sidak’s multiple comparisons test (α = 0.05) showed that VITAM20 exhibited significantly higher stabilization percentages than diclofenac (group C) at all concentrations (200 μg/mL–1,000 μg/mL), with adjusted p-values <0.0001. The mean difference in stabilization ranged from 7.65% to 28.04%, indicating a stronger anti-inflammatory effect of Kurthia gibsonii VITAM20 compared to that of diclofenac.

At the tested concentrations, Kurthia gibsonii VITAM20 exhibited protection of 69.92% ±1.08% under heated conditions and 90.78% ± 1.03% in the absence of heat, signifying a significant degree of membrane stabilization (Figure 4A). Conversely, the reference drug diclofenac showed protection of 83.83% ± 1.23% (upon heating) and 83.01% ± 0.72% (without heating). A two-way repeated-measures ANOVA showed significant effects of treatment, heating, and their interaction. Both VITAM20 and diclofenac differed significantly in anti-inflammatory activity (p = 0.0112), while unheated samples exhibited significantly higher activity than heated samples (p = 0.0095). The significant interaction (p = 0.0122) indicates that heating affected Kurthia gibsonii VITAM20 more strongly than diclofenac, indicating that Kurthia gibsonii VITAM20 is comparatively less heat-stable.

FIGURE 4.

Bar chart figure compares two sets of experimental results. Panel a displays percentage of protection between heated and unheated samples for VITAM20 and Diclofenac in an HRBC membrane stabilization assay, with VITAM20 showing significant difference (****) and Diclofenac showing no significance (ns). Panel b visualizes inhibition of protein denaturation using egg albumin across five sample concentrations (200 to 1000 micrograms per milliliter) for VITAM20 and Diclofenac. Both samples exhibit high and similar inhibition, with several significance markers noted (****, ***), and some comparisons labeled as not significant (ns).

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(a) Comparison of heat-induced and non-heat-induced hemolysis between Kurthia gibsonii VITAM20 crude enzyme extract and the standard drug diclofenac. (b) Effect of crude enzyme extract from Kurthia gibsonii VITAM20 on egg albumin denaturation compared with that of the standard diclofenac. All experiments were performed in triplicate (n = 3), and data are presented as the mean values.

The experiment evaluating the denaturation of egg albumin was conducted to analyze the anti-inflammatory characteristics of Kurthia gibsonii VITAM20 at different concentrations, with diclofenac acting as the reference. Protein denaturation was strongly increased in Kurthia gibsonii VITAM20 in a concentration-dependent manner, such as 80.94% ± 0.57% (200 μg/mL), 82.58% ± 0.43% (400 μg/mL), 82.7% ± 0.32% (600 μg/mL), 83.41% ± 0.32% (800 μg/mL), and 84.47% ± 0.39% (1000 μg/mL). Simultaneously, diclofenac demonstrated 74.82% ± 1.08%, 78.58% ± 0.47%, 78.35% ± 0.44%, 82.23% ± 0.44%, and 83.29% ± 0.69% increase (Figure 4B). Sidak’s multiple comparisons test found that VITAM20 showed significantly higher activity than diclofenac at 200 μg/mL, 400 μg/mL, and 600 μg/mL (p < 0.0001, p = 0.0003, and p = 0.0001), respectively, but no considerable difference at 800 μg/mL and 1000 μg/mL (p = 0.5085 and p = 0.2565). This demonstrates that the advantage of VITAM20 over diclofenac is prominent only at lower concentrations.

3.6. Antioxidant activity results

The antioxidant effect of the crude extract of Kurthia gibsonii VITAM20 was systematically assessed through the DPPH radical scavenging assay. The DPPH assay demonstrated the test isolate’s ability to eliminate free radicals. All experiments were performed in triplicate, and the results are expressed as the mean ± standard deviation. The cell free supernatant sample of Kurthia gibsonii VITAM20 showed 64.27% ± 0.46% (500 μg/mL) of scavenging, respectively. Ascorbic acid (100 μg/mL), the positive control, showed 91.40% ± 0.46% free radical scavenging. Ascorbic acid exhibited significantly higher radical scavenging activity than Kurthia gibsonii VITAM20 (p = 0.0081). Increasing the concentration significantly enhanced the antioxidant activity in both the samples (p = 0.0005). A significant interaction between the sample type and concentration was also observed (p = 0.0006), indicating that the response to concentration differed between Kurthia gibsonii VITAM20 and ascorbic acid. Overall, ascorbic acid demonstrated superior antioxidant potential than Kurthia gibsonii VITAM20. The concentration required to inhibit 50% of DPPH radicals (IC50) was calculated by linear interpolation of the dose–response curve. The IC50 value of the VITAM20 extract was determined to be 286.5 μg/mL, whereas ascorbic acid exhibited a substantially lower IC50 value, indicating superior antioxidant potency. These findings indicate that the VITAM20 extract shows moderate free radical scavenging activity in a dose-dependent manner (Figure 5).

FIGURE 5.

Bar chart titled DPPH Assay showing DPPH scavenging percentages for VITAM20 and ascorbic acid at three concentrations: 100, 250, and 500 micrograms per milliliter. Ascorbic acid exhibits higher scavenging activity across all concentrations compared to VITAM20, with statistical significance indicated by asterisks.

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Antioxidant activity of Kurthia gibsonii VITAM20 extract measured by DPPH radical scavenging and expressed as the mean ± SD (n = 3).

3.7. Anti-bacterial activity

The crude enzyme samples formed a 13-mm sized zone of inhibition against Staphylococcus aureus and Listeria monocytogenes, respectively (Table 1), and the zone size of the positive control was 25 mm.

TABLE 1.

Antibacterial activity of the crude enzyme extract from Kurthia gibsonii VITAM20 against selected bacterial strains.

Sl. No Concentration of the sample (µg/mL) Test organism Zone of inhibition (mm)
1 100 Listeria monocytogenes No zone
2 100 Staphylococcus aureus 13 mm
3 100 Pseudomonas aeruginosa No zone
4 100 Escherichia coli 13 mm
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The antibacterial activity of the crude enzyme extract against Listeria monocytogenes and Staphylococcus aureus was evaluated using MIC. All experiments were performed in triplicate (n = 3), and the results are presented as the mean optical density (OD ± SD). A concentration-dependent inhibitory effect was observed against both test organisms. For L. monocytogenes, the mean OD decreased from 0.339 ± 0.001 at 9.0625 μg/mL to 0.165 ± 0.005 at 250 μg/mL. Similarly, for S. aureus, it exhibited a reduction from 0.375 ± 0.004 to 0.212 ± 0.001 over the same concentration range. Against L. monocytogenes, the percentage inhibition ranged from 61.91% at 9.0625 μg/mL to 81.46% at 250 μg/mL. Likewise, for S. aureus, it showed inhibition ranging from 60.36% at 9.0625 μg/mL to 77.82% at 250 μg/mL. The MIC was determined to be 250 μg/mL (0.25 mg/mL) for both pathogens. At this concentration, the crude enzyme extract inhibited the growth of L. monocytogenes and S. aureus by 81.46% and 77.82%, respectively. The MIC50, representing the concentration required to inhibit 50% of bacterial growth, was calculated as 72.5 μg/mL for both bacterial strains. Since 90% inhibition was not achieved within the tested concentration range, the MIC90 was estimated to be greater than 250 μg/mL for both organisms.

The positive control exhibited OD values of 0.240 for L. monocytogenes and 0.159 for S. aureus, whereas the sterility and solvent controls showed negligible absorbance values (<0.07), confirming the validity and reliability of the assay.

4. Discussion

Bacteria play an important role, including the synthesis of antimicrobial substances and various metabolites. Various studies have shown that the most effective strategy to safeguard human beings from the consequences of oxidative reactions is to preclude such reactions, which can be achieved through the utilization of antioxidants. In recent years, antioxidants that occur in nature and are extracted from biological origins have garnered notable interest as a sensible replacement for synthetic antioxidants (Faraki and Rahmani, 2020).

Considering the pharmaceutical significance and industrial demand of this enzyme, the current study was concentrated on the identification of a strain with bioactive potential. The whole-genome characterization of the bacterial isolate revealed that it was the Kurthia gibsonii VITAM20 strain with 2,970,044 base pairs (bp), including 6,973 predicted genes. The GC content of the strain VITAM20 (36.5%) was within the reported range for bacterial genomes (13%–75%), indicating that the strain possesses a typical genomic base composition that is consistent with previously characterized bacteria (Mesbah et al., 2011). In previous studies, K. sibirica NBRC 101530 possessed the largest genome size (3.4 Mb), whereas K. gibsonii M6 exhibited the smallest genome (2.8 Mb). The highest GC content (39.50%) was observed in K. massiliensis JC30, while the lowest (36.00%) was reported in K. sibirica NBRC 101530. In comparison, our Kurthia gibsonii VITAM20 exhibited a GC content of 36.5% and a genome size of 2.9 Mb, which was within the range reported for the genus (Roy et al., 2025).

The growth curve follows the lag (0 h–12 h), exponential (12 h–30 h), stationary (30 h–38 h), and decline (>38 h) stages. This agrees with normal microbial growth. Compared to other studies, the lag phase here is slightly longer, indicating slower adaptation of the cells to the medium. The exponential phase shows moderate growth rather than steep increases, indicating possible environmental or nutrient limitations. The stationary phase peak and slight fluctuation are consistent with reports of metabolic adjustments in dense cultures, while the sharp decline phase indicates nutrient depletion and accumulation of toxic byproducts, which occurs more gradually in some studies (Jones et al., 2023; Kumakura et al., 2023).

In the Planococcaceae family, Kurthia has an evolutionary relationship with some species such as Lactobacillus, Bacillus, Staphylococcus, and Streptococcus (Paul et al., 2022). The strain Kurthia gibsonii VITAM20 was isolated from a soil sample and was identified with significant bioactive properties. The analysis revealed a diverse collection of genes linked to anti-inflammatory and antioxidant functions within the bacterial strain. In addition to this, the presence of both terpene compounds and their precursors was identified by antiSMASH 7.0.

Genes associated with anti-inflammatory properties such as eps (95.455%), NEMO (100%), spa (100%), and spaB (90.625%) were identified within the genome. Especially, luxS and NEMO are important for the development of quorum sensing and the regulation of NF-κB signaling. The genes eps and slpA are substantial in the structural and interactive components of bacterial extracellular polysaccharides and surface layers and are valued for their effect on decreasing inflammatory responses. The presence of the spa and spaB genes further validates the strain’s potential in the modulation of immune responses.

NEMO is required for the preservation of epithelial integrity and the regulation of intestinal inflammation through NF-κB signaling and the modulation of apoptotic processes, thereby serving as a prospective therapeutic target in the modulation of inflammatory diseases (Tas et al., 2006; Nenci et al., 2007). Genes linked with antioxidant properties, such as gshR3 (91.304%), gshR4 (100%), KatA (100%), CAT (92.308%), dps (95.238%), oxyR (100%), KatA_pseudo (88.462%), sodA (100%), SodM (76.471%), perR (74.041% with the maximum bit score of 126), and trxA (77.778%), specify a robust system for the detoxification of reactive oxygen species. These genes encode enzymes such as superoxide dismutase, catalases, and thioredoxins that together function to maintain the cellular redox balance and oxidative damage. The completeness and characteristic diversity in this antioxidant gene set recommend the strain’s strong ability to withstand oxidative stress and support host health through antioxidative mechanisms. Kurthia gibsonii has been reported to exhibit antioxidant potential as an adaptive response to environmental stress. Certain strains, for example, K. gibsonii TYL-A1, showed antioxidant activity through the production of enzymes such as superoxide dismutase and catalase (Wang et al., 2024).

In an earlier study, hepatocytes and fibroblasts obtained from Tg (CAT) (+/+) mice demonstrated an increased resistance to cell death induced by hydrogen peroxide (Chen et al., 2004). The thioredoxin (Trx) system is needed for electron transfer to thiol-dependent peroxidases, supporting the effective eradication of reactive oxygen. The antioxidant function of Trx was also evidenced through its participation in DNA and protein repair mechanisms by inhibiting ribonucleotide reductase and methionine sulfoxide reductases, in addition to modulating several redox-sensitive transcription factors. Furthermore, Trx systems played a vital role in immune responses, cell death, and viral infection processes through interactions with thioredoxin-interacting proteins (Lu and Holmgren, 2014). The DNA-binding protein derived from starved cells (Dps) conferred protection against oxidative stress during growth in the exponential phase and additionally conferred resistance to UV and gamma irradiation, along with protection against harmful effects of iron and copper, thermal pressure, and acid–base shock (Nair and Finkel, 2004). Glutathione reductase (GSR), a pivotal enzyme within the glutathione antioxidant defense mechanism, catalyzes the reduction of oxidized glutathione (GSSG) to its reduced form (GSH), thus preserving the intracellular redox state to guard cellular defense against oxidative damage (Han et al., 2017). In another study, Pseudomonas aeruginosa OxyR was recognized as an important transcriptional regulator of the katA gene, capable of finding H2O2 through conserved cysteine residues and undergoing many oxidation and activation states in vivo (Heo et al., 2010). Together, the important enzymes capable of antioxidant resistance such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), were required for the dismutation of superoxide radicals (O2-) and hydrogen peroxide (H2O2). SOD inhibits the synthesis of peroxynitrite and thus maintains physiologically significant levels of nitric oxide, which is necessary for vasodilation neurotransmission and inflammatory reactions (Jomova et al., 2024). The antiSMASH analysis further revealed the presence of biosynthetic gene clusters linked with terpenes and their precursors. Terpenes and their precursors emphasize the strain’s capacity to produce these important secondary metabolites. The bioactive potential of terpenes is known for their diversity, including antimicrobial, anti-inflammatory, and antioxidant potential, which could increase the genetic makeup of the strain to improve the health benefits. The presence of terpene biosynthesis pathways significantly enhances the therapeutic potential of this bacterial strain, providing biochemical importance for the observed genetic traits related to immune modulation and resistance to oxidative stress. Kurthia gibsonii, isolated from paneer, and the characterization of bacteriocin produced by this bacterium was well-explained in a previous study (Chauhan and Samant, 2022).

Denaturation of tissue proteins has been marked as a key component of the disease pathways tied to arthritis and inflammation. Research findings reveal that when proteins are denatured, autoantigens can develop in various types of arthritis. An increase in the absorbance of the protein hydrolysate and the reference drug, in comparison to the control group, signified the stabilization of albumin protein. Due to the structural resemblance between erythrocyte and lysosomal membranes, the HRBC assay was utilized for the in vitro study of anti-inflammatory potential. The stability of lysosomal membranes is significant in the regulation of inflammatory responses by inhibiting the secretion of bactericidal and proteolytic enzymes from activated neutrophils, which cause tissue damage and inflammatory processes (Wang et al., 2021; Rivera-Jiménez et al., 2022).

The in vitro studies on Kurthia gibsonii VITAM20 showed prominent antibacterial, anti-inflammatory, and antioxidant properties, thereby strengthening its potential therapeutic applications. The membrane stabilization assay showed that Kurthia gibsonii VITAM20 exhibited 90.78% protection under non-heated conditions and 69.92% protection under heated conditions. This confirmed that the isolate efficiently stabilizes the lysosomal membrane’s integrity by stopping the release of inflammatory mediators. The protein denaturation assay using egg albumin further confirmed the anti-inflammatory potential of Kurthia gibsonii VITAM20, demonstrating a concentration-dependent increase of protein denaturation across five different concentrations, with an increase of activity varying from 80.94% to 84.47%. Similar studies using egg albumin denaturation assays have validated this assay as a reliable approach for evaluating the anti-inflammatory potential of both natural and microbial agents, displaying a strong relationship with NSAIDs such as diclofenac, which served as the positive control. Previous studies have confirmed that the crude extract of S. marcescens VS56 exhibits significant dose-dependent HRBC membrane stabilization and shows the anti-inflammatory potential of 94.6%, which is comparable to the standard drug diclofenac (94.6%) (Nair and Subathra Devi, 2024). Although some studies have reported that HRBC protection by diclofenac differs depending on the concentration, incubation conditions, and the type of hemolysis inducer used (Parameswari et al., 2019).

The present study demonstrated that the Kurthia gibsonii VITAM20 extract possesses considerable antioxidant activity, as evidenced by its concentration-dependent DPPH radical scavenging ability. The increase in scavenging activity with increasing concentration indicates the presence of bioactive compounds capable of donating hydrogen atoms or electrons to neutralize free radicals. Although the antioxidant activity of the extract was lower than that of ascorbic acid, the observed IC50 value of 286.5 μg/mL confirms its moderate antioxidant potential.

In terms of antibacterial efficiency, the crude enzyme extracts derived from Kurthia gibsonii VITAM20 exhibited inhibitory activity against Staphylococcus aureus and Listeria monocytogenes, producing zones of inhibition of 13 mm. Although the zones of inhibition were moderate, these findings were significant considering previous studies that reported similar antimicrobial properties of bacterial isolates against these pathogens. The crude enzyme extract exhibited potent antibacterial activity against both Listeria monocytogenes and Staphylococcus aureus, as evidenced by the concentration-dependent reduction in bacterial growth. The identical MIC value of 250 μg/mL against both organisms indicates broad-spectrum efficacy against Gram-positive foodborne pathogens.

The higher inhibition observed against L. monocytogenes (81.46%) compared to S. aureus (77.82%) indicates a slightly greater susceptibility of Listeria to the bioactive compounds present in the crude extract. The MIC value obtained in the present study falls within the range considered indicative of moderate-to-strong antibacterial activity for crude natural extracts. Extracts exhibiting MIC values below 625 μg/mL are generally regarded as promising antimicrobial candidates (Zouine et al., 2024). Comparable antibacterial activities have been reported for extracellular metabolites and crude extracts from bacterial isolates. For instance, the ethyl acetate extract of Bacillus amyloliquefaciens VJ-1 exhibited a MIC of 25 μg/mL against Staphylococcus aureus, demonstrating the remarkable antimicrobial potential of Bacillus-derived bioactive compounds (Kadaikunnan et al., 2015). Likewise, cell-free supernatants from Bacillus subtilis KU43 effectively inhibited Listeria monocytogenes, with a MIC of 12.5% (v/v), further supporting the susceptibility of Listeria to Bacillus-derived antimicrobial metabolites (Park et al., 2023). The inability to achieve MIC90 within the tested concentration range indicates that concentrations above 250 μg/mL may be required for near-complete bacterial eradication. Overall, these results highlight the promising antibacterial potential of the crude enzyme extract as a natural antimicrobial agent. Further purification, characterization, and mechanistic studies are required to identify the active constituents responsible for the observed inhibitory activity and evaluate their potential applications.

The present study was limited to a preliminary evaluation using the crude supernatant, and the specific bioactive compounds responsible for the observed activity were not purified or characterized. Further validation through in vivo studies will be conducted in future studies. Overall, these findings indicated that the extract showed anti-inflammatory activity comparable to that of the standard drug, diclofenac, while also exhibiting moderate antibacterial activity against clinically significant pathogens. This dual functionality was consistent with previous studies on this isolate and its derived compounds, highlighting its potential as a promising source for the development of novel anti-inflammatory, antibacterial, and antioxidant agents.

5. Limitations of the study

The present study is preliminary in nature and has certain limitations. Genome mining was based primarily on sequence similarity and requires further validation through phylogenetic, comparative genomic, and gene expression analyses. Although significant biological activities were observed, the specific bioactive compounds responsible for these effects were not purified or structurally characterized. Furthermore, the anti-inflammatory effects were evaluated only through in vitro assays, and detailed mechanistic and in vivo studies are needed. These limitations will be addressed in future studies.

6. Conclusion

The presence of anti-inflammatory and antioxidant genes, combined with the terpene biosynthetic capability, underscored the Kurthia gibsonii VITAM20 strain’s functional potential for bioactive compound production. These genomic features indicated a strong potential for managing inflammation and oxidative stress-related conditions, requiring further experimental validation to confirm their efficacy and bioactivity in vivo. To the best of our knowledge, this was the first report to reveal the anti-inflammatory and antioxidant properties of Kurthia gibsonii.

Acknowledgements

The authors would like to thank the Vellore Institute of Technology for providing them the required facilities to pursue this research.

Funding Statement

The author(s) declared that financial support was not received for this work and/or its publication.

Footnotes

Edited by: Arun Karnwal, Graphic Era University, India

Reviewed by: Abdel Moneim Elhadi Sulieman, University of Hail, Saudi Arabia

Sugam Gupta, Tula’s Institute, India

Data availability statement

The original contributions presented in the study are publicly available. This data can be found here: BioProject: PRJNA1238542 (https://www.ncbi.nlm.nih.gov/bioproject/PRJNA1238542); BioSample: SAMN47475936 (https://www.ncbi.nlm.nih.gov/biosample/SAMN47475936); SRA: SRR35894991 (https://www.ncbi.nlm.nih.gov/sra/SRR35894991); Genome accession: CP185371.1 (https://www.ncbi.nlm.nih.gov/nuccore/CP185371.1).

Ethics statement

The study protocol involving human blood samples and all the related in vitro assays was reviewed and approved by the Vellore Institute of Technology Institutional Ethics Committee for Human Studies (Approval No: VIT/IECH/2025/16 IECH/15 February 2025/19). The studies were conducted in accordance with the local legislation and institutional requirements. The participants provided their written informed consent to participate in this study.

Author contributions

AA: Investigation, Software, Visualization, Writing – original draft, Conceptualization, Formal Analysis. MV: Conceptualization, Supervision, Methodology, Visualization, Investigation, Formal Analysis, Writing – review and editing, Project administration.

Conflict of interest

The author(s) declared that this work was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Generative AI statement

The author(s) declared that generative AI was not used in the creation of this manuscript.

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Supplementary material

The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fphar.2026.1799206/full#supplementary-material

SUPPLEMENTARY IMAGE 1

Determination of the minimum inhibitory concentration (MIC) of the crude enzyme extract from Kurthia gibsonii VITAM20 against Staphylococcus aureus and Listeria monocytogenes using the broth microdilution method.

Table1.docx (21.5KB, docx)
Image1.tif (1.2MB, tif)
DataSheet1.pdf (367.6KB, pdf)

References

  1. Akwayamai John J., Henry Bulama G. (2020). The in-vitro anti-inflammatory and anti-haemolytic activity of the stem bark methanol extract of khaya senegalensis. World J. Pharm. Res. 9, 2242. 10.20959/wjpr20206-17736 [DOI] [Google Scholar]
  2. Al-Mijalli S. H., El Hachlafi N., Jeddi M., Abdallah E. M., Assaggaf H., Qasem A., et al. (2023). Unveiling the volatile compounds and antibacterial mechanisms of action of Cupressus sempervirens L., against Bacillus subtilis and Pseudomonas aeruginosa . Biomed. Pharmacother. 167, 115609. 10.1016/j.biopha.2023.115609 [DOI] [PubMed] [Google Scholar]
  3. Ali K. A., Maity A., Roy S. D., Das Pramanik S., Pratim Das P., Shaharyar M. A. (2023). Insight into the mechanism of steroidal and non-steroidal anti-inflammatory drugs. How Synthetic Drugs Work: Insights into Molecular Pharmacology of Classic and New Pharmaceuticals , 61–94. 10.1016/B978-0-323-99855-0.00004-X [DOI] [Google Scholar]
  4. Chaiya P., Senarat S., Phaechamud T., Narakornwit W. (2022). In vitro anti-inflammatory activity using thermally inhibiting protein denaturation of egg albumin and antimicrobial activities of some organic solvents. Mater. Today Proc. 65, 2290–2295. 10.1016/J.MATPR.2022.04.916 [DOI] [Google Scholar]
  5. Chauhan A., Samant S. A. (2022). Isolation and identification of Kurthia gibsonii from paneer and study its antibacterial activity against intestinal pathogens. Int. J. Health Sci. 6 (S5), 10457–10466. 10.53730/IJHS.V6NS5.11269 [DOI] [Google Scholar]
  6. Chen X., Liang H., Remmen H. V. J. V.-A., Liang H., Van Remmen H., Vijg J., et al. (2004). Catalase transgenic mice: characterization and sensitivity to oxidative stress. Archives Biochemistry Biophysics, 2004•Elsevier 422, 197–210. 10.1016/J.ABB.2003.12.023 [DOI] [PubMed] [Google Scholar]
  7. Clinical and Laboratory Standards Institute (CLSI). (2010). Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standard, 8th Edition. CLSI document M07-A8 (Wayne, PA: Clinical and Laboratory Standards Institute). [Google Scholar]
  8. Faraki A., Rahmani F. (2020). The antioxidant activity of lactic acid bacteria and probiotics: a review. Journal Food Saf. Hyg. 6 (4), 168–182. 10.18502/jfsh.v6i4.7563 [DOI] [Google Scholar]
  9. Gusev E., Zhuravleva Y. (2022). Inflammation: a new look at an old problem. Int. J. Mol. Sci. 23, 4596. 10.3390/IJMS23094596 [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Han C., Kim M. J., Ding D., Park H. J., White K., Walker L., et al. (2017). GSR is not essential for the maintenance of antioxidant defenses in mouse cochlea: possible role of the thioredoxin system as a functional backup for GSR. PLoS One 12, 12. 10.1371/JOURNAL.PONE.0180817 [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hasan M. M., Islam M. E., Hossain M. S., Akter M., Rahman M. A. A., Kazi M., et al. (2024). Unveiling the therapeutic potential: evaluation of anti-inflammatory and antineoplastic activity of Magnolia champaca Linn’s stem bark isolate through molecular docking insights. Heliyon 10, e22972. 10.1016/J.HELIYON.2023.E22972 [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Heo Y.-J., Chung I.-Y., Cho W.-J., Lee B.-Y., Kim J.-H., Choi K.-H., et al. (2010). The major catalase gene (katA) of pseudomonas aeruginosa PA14 is under both positive and negative control of the global transactivator OxyR in response to, 192, 381–390. 10.1128/JB.00980-09 [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jomova K., Alomar S. Y., Alwasel S. H., Nepovimova E., Kuca K., Valko M., et al. (2024). Several lines of antioxidant defense against oxidative stress: antioxidant enzymes, nanomaterials with multiple enzyme-mimicking activities, and low-molecular-weight. Arch Toxicol. 98, 1323–1367. 10.1007/S00204-024-03696-4 [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jones E., Derrick J., Nisbet R., Reports W. L.-S., Jones comE. W., Derrick J., et al. (2023). First-passage-time statistics of growing microbial populations carry an imprint of initial conditions. Nature 13, 21340. 10.1038/s41598-023-48726-w [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Joseph J., Bindhu A. R., Aleykutty N. A. (2013). In vitro and in vivo antiinflammatory activity of Clerodendrum paniculatum linn. Leaves. Indian J. Pharm. Sci. 75, 376–379. 10.4103/0250-474X.117428 [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kadaikunnan S., Rejiniemon S. S., Khaled J. M., Alharbi N. S., Mothana R., Kadaikunnan S. S., et al. (2015). In-vitro antibacterial, antifungal, antioxidant and functional properties of Bacillus amyloliquefaciens. R MothanaAnnals Clinical Microbiology Antimicrobials 14, 9. 10.1186/S12941-015-0069-1 [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kumakura D., Yamaguchi R., Hara A., Nakaoka S. (2023). Disentangling the growth curve of microbial culture. J. Theor. Biol. 573, 111597. 10.1016/j.jtbi.2023.111597 [DOI] [PubMed] [Google Scholar]
  18. Liu S., Deng Z., Chen K. S. J.-M., Deng Z., Chen K., Jian S., et al. (2022). Cartilage tissue engineering: from proinflammatory and anti-inflammatory cytokines to osteoarthritis treatments. Mol. Med. Rep. 25 (3), 1–15. 10.3892/mmr.2022.12615 [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lu J., Holmgren A. (2014). The thioredoxin antioxidant system. Free Radical Biology Medicine 66, 75–87. 10.1016/J.FREERADBIOMED.2013.07.036 [DOI] [PubMed] [Google Scholar]
  20. Melnikova O. V., Ivanenko T. V., Grekova T. A., Kadzharyan Y. V. (2018). “Pathogenesis of inflammation and peripheral circulation disturbances module No 1,” in General Pathophysiology Submodule 2. Typical Pathological Processes. [Google Scholar]
  21. Mesbah N. M., Whitman W. B., Mesbah M. (2011). Determination of the G+C content of prokaryotes. Methods Microbiol. 38, 299–324. 10.1016/B978-0-12-387730-7.00014-0 [DOI] [Google Scholar]
  22. Nair S., Finkel S. E. (2004). Dps protects cells against multiple stresses during stationary phase. J. Bacteriology 186, 4192–4198. 10.1128/JB.186.13.4192-4198.2004 [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nair S. R., Subathra Devi C. (2024). Bioprospecting of serratiopeptidase-producing bacteria from different sources. Front. Microbiol. 15, 1382816. 10.3389/FMICB.2024.1382816/FULL [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Nenci A., Becker C., Wullaert A., Gareus R., van Loo G., Danese S., et al. (2007). Epithelial NEMO links innate immunity to chronic intestinal inflammation. Nature 446, 557–561. 10.1038/NATURE05698 [DOI] [PubMed] [Google Scholar]
  25. Parameswari P., Devika R., Vijayaraghavan P. (2019). In vitro anti-inflammatory and antimicrobial potential of leaf extract from Artemisia nilagirica (clarke) pamp. Saudi J. Biol. Sci. 26 (3), 460–463. 10.1016/j.sjbs.2018.09.005 [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Park Y., Kim Y., Yu H., Lee N., Control H. P.-F. (2023). Cell-Free Supernatants of Bacillus subtilis and Bacillus Polyfermenticus Inhibit Listeria monocytogenes Biofilm Formation. Available online at: https://www.sciencedirect.com/science/article/pii/S0956713522005801 (Accessed April 30, 2026). [Google Scholar]
  27. Paul M. K., Mini K., Antony A. C., Mathew J. (2022). Deproteinization of shrimp shell waste by Kurthia gibsonii Mb126 immobilized chitinase. J. Pure Appl. Microbiol. 16, 909–923. 10.22207/JPAM.16.2.11 [DOI] [Google Scholar]
  28. Pourramezan Z., Oloomi M., Kasra-Kermanshahi R. (2020). Antioxidant and anticancer activities of Lactobacillus Hilgardii strain AG12a. Int. J. Prev. Med. 11, 132. 10.4103/IJPVM.IJPVM_307_19 [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rivera-Jiménez J., Berraquero-García C., Pérez-Gálvez R., García-Moreno P. J., Espejo-Carpio F. J., Guadix A., et al. (2022). “Peptides and protein hydrolysates exhibiting anti-inflammatory activity: sources, structural features and modulation mechanisms,” in Rsc.Org. 10.1039/D2FO0+2223K.+S [DOI] [PubMed] [Google Scholar]
  30. Roy V., Pal M. S., Pal A. (2025). Analysis of heavy metal tolerance and genomics in an Indigenous kurthia strain from Kulik river reveals multi-metal resistance and dominance of selection pressure on codon usage patterns. Archives Microbiol. 207 (3), 57. 10.1007/s00203-025-04255-w [DOI] [PubMed] [Google Scholar]
  31. Sohail R., Mathew M., Patel K. K., Reddy S. A., Haider Z., Naria M., et al. (2023). Effects of Non-steroidal Anti-inflammatory Drugs (Nsaids) and Gastroprotective Nsaids on the Gastrointestinal Tract: A Narrative Review. 10.7759/cureus.37080 [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tas S. W., Vervoordeldonk M. J., Hajji N., May M. J., Ghosh S., Tak P. P., et al. (2006). Local treatment with the selective IκB kinase β inhibitor NEMO-Binding domain peptide ameliorates synovial inflammation. Arthritis Res. Ther. 8 (4), R86. 10.1186/AR1958 [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tumilaar S. G., Hardianto A., Dohi H., Kurnia D. (2024). A comprehensive review of free radicals, oxidative stress, and antioxidants: overview, clinical applications, global perspectives, future directions, and mechanisms of antioxidant activity of flavonoid compounds. J. Chem. 2024, 5594386. 10.1155/2024/5594386 [DOI] [Google Scholar]
  34. Valgas C., De Souza S. M., Smânia E. F. A., Smânia A. (2007). Screening methods to determine antibacterial activity of natural products. Braz. J. Microbiol. 38, 369–380. 10.1590/S1517-83822007000200034 [DOI] [Google Scholar]
  35. Wang B., Wu L., Chen J., Dong L., Chen C., Wen Z., et al. (2021). Metabolism pathways of arachidonic acids: mechanisms and potential therapeutic targets. Signal Transduct. Target. Ther. 6, 94. 10.1038/S41392-020-00443-W [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wang Y., Zhao C., Zhao B., Duan X., Hao P., Liang X., et al. (2024). Transcriptome of Kurthia gibsonii TYL-A1 revealed the biotransformation mechanism of tylosin. Microorganisms 12 (12), 2597. 10.3390/microorganisms12122597 [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Zouine N., Ghachtouli N. E., Abed S. E., African S. K.-S. (2024). A comprehensive review on medicinal plant extracts as antibacterial agents: factors, mechanism insights and future prospects. Scientific African 26, e02395. 10.1016/j.sciaf.2024.e02395 [DOI] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

SUPPLEMENTARY IMAGE 1

Determination of the minimum inhibitory concentration (MIC) of the crude enzyme extract from Kurthia gibsonii VITAM20 against Staphylococcus aureus and Listeria monocytogenes using the broth microdilution method.

Table1.docx (21.5KB, docx)
Image1.tif (1.2MB, tif)
DataSheet1.pdf (367.6KB, pdf)

Data Availability Statement

The original contributions presented in the study are publicly available. This data can be found here: BioProject: PRJNA1238542 (https://www.ncbi.nlm.nih.gov/bioproject/PRJNA1238542); BioSample: SAMN47475936 (https://www.ncbi.nlm.nih.gov/biosample/SAMN47475936); SRA: SRR35894991 (https://www.ncbi.nlm.nih.gov/sra/SRR35894991); Genome accession: CP185371.1 (https://www.ncbi.nlm.nih.gov/nuccore/CP185371.1).

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