JIPK.
Volume 17 No 3 October 2025 Sinta 1 (Decree No: 158/E/KPT/2.
e-ISSN:2528-0759.
p-ISSN:2085-5842 Available online at https://e-journal.
id/JIPK JIPK
(JURNAL ILMIAH PERIKANAN DAN KELAUTAN)
Scientific Journal of Fisheries and Marine Research Article In vitro and In silico Antibacterial Activity of Centella asiatica Leaves Bioactive Compounds Against Fish Pathogenic Bacteria Septyan Andriyanto 1,4* .
Maftuch 1 .
Sri Andayani 1 Hessy Novita 2 , and Muhammad Nursid 3 .
Nunak Nafiqoh 2 .
Lila Gardenia 2 Department of Aquaculture.
Faculty of Fisheries and Marine Science.
Brawijaya University.
Malang, 65145.
Indonesia Research Centre of Veterinary Science.
National Research and Innovation Agency (BRIN).
Cibinong.
Bogor, 16915.
Indonesia Research Center for Marine and Land Water Bioindustry.
National Research and Innovation Agency.
Lombok, 83352.
Indonesia Research Center for Conservation of Marine and Inland Water Resources.
National Research and Innovation Agency.
Cibinong.
Bogor, 16915.
Indonesia
Abstract
ARTICLE INFO
Received: April 24, 2025 Accepted: June 19, 2025 Published: July 16, 2025 Available online: Sept 27, 2025 *) Corresponding author:
E-mail: septian09@student.
Keywords:
GC-MS
Molecular docking DNA gyrase Crude Extracts MIC This is an open access article under the CC BY-NC-SA license .
ttps://creativecommons.
org/licenses/by-nc-sa/4.
Antimicrobial agents are crucial for managing bacterial infections in fish Centella asiatica is a medicinal plant recognised for its diverse bioactive compounds with important antibacterial properties.
The present study aimed to investigate the antibacterial activity of C.
asiatica leaves bioactive compounds on fish pathogenic bacteria using an In vitro and In silico approach.
The maceration method was used to extract bioactive compounds from C.
asiatica leaves and was identified using Gas Chromatography-Mass Spectrometry (GC-MS).
In vitro analysis of antibacterial activity was evaluated using the minimum inhibitory concentration method.
While In silico molecular docking is applied alongside assessing LipinskiAos rules of five, as well as absorption, distribution, metabolism, excretion, and toxicity properties.
The result of the GC-MS examination of the asiatica leaf extracts identified 53 bioactive compounds.
In vitro studies showed antibacterial efficacy of leaf extracts against fish pathogenic bacteria (Streptococcus agalactiae.
Bacillus subtilis, and Staphylococcus aureu.
with minimum inhibitory concentration values of 12,5 mg/ml.
In silico molecular docking analysis showed that several bioactive compounds have the potential to be DNA gyrase inhibitors.
Compound 13-Hexyloxacyclotridec-10-en-2-one has the highest inhibition with binding energy of Oe7,4 Kcal/mol compared to ciprofloxacin as drug standard with a binding energy value Oe7,3 Kcal/mol.
The following compound is gamma.
-Muurolene (Oe6,7 Kcal/mo.
Copaene (Oe6,6 Kcal/mo.
and Humulene (Oe6,6 Kcal/mo.
These results suggest that bioactive compounds of C.
asiatica leaves extracts hold promise as potential antibacterial agents for treating fish pathogenic bacteria infections.
Cite this as: Andriyanto.
Maftuch.
Andayani.
Nafiqoh.
Gardenia.
Novita.
, & Nursid.
In vitro and In silico antibacterial activity of Centella asiatica leaves extracts against fish pathogenic bacteria.
Jurnal Ilmiah Perikanan dan Kelautan, 17.
:591-607.
https://doi.
org/10.
20473/jipk.
Copyright A2025 Faculty of Fisheries and Marine Universitas Airlangga Andriyanto et al.
/ JIPK, 17.
:591-607
Introduction Pathogenic bacteria increasingly impact fish culture, as higher population densities can exacerbate disease outbreaks.
Effective antimicrobial agents are essential for addressing bacterial infections in fish culture frameworks.
Recent studies show that traditional medicinal plant extracts, particularly Centella asiatica, may help manage bacterial pathogens in aquaculture (Si et al.
, 2023.
Jenitha, 2.
Centella asiatica, also known as Pegagan in Indonesia, is a notable medicinal plant known for its many bioactive components demonstrating remarkable antibacterial activities.
This distinctive herb, part of the Apiaceae family, has been traditionally utilized in several civilizations, especially in Asia, to treat numerous diseases.
The medicinal effectiveness of C.
asiatica is largely attributed to its wide range of bioactive substances, notably terpenoids, saponins, flavonoids, tannins, alkaloid, and steroids, which play a significant role in enhancing its bioactivity, particularly its antibacterial properties (Liu et al.
, 2020.
Magaya et al.
, 2020.
Yusof et al.
Akkol et al.
, 2021.
Mohapatra et al.
, 2.
This plant contains several important triterpenes, including asiaticoside, madecassoside, asiatic acid, and madecassic acid, all of which have recognized health benefits, especially their antibacterial effects (Sun et al.
, 2020.
Tripathy et al.
, 2022.
Wei et al.
Wang et al.
, 2024.
Previous studies reported strong antibacterial effects of C.
asiatica extracts against common fish pathogens like Vibrio harveyi and Aeromonas hydrophila, which cause significant economic losses in aquaculture (Rukisah et al.
, 2.
Leaf extracts and endophytic fungi associated with C.
asiatica have been shown strong antimicrobial activity in aquaculture and can suppress the growth of fish and shellfish pathogenic bacteria (Shankar and Sathiavelu, 2.
Centella asiatica extracts are beneficial not only for pathogen control but also for improving biosecurity in aquaculture systems.
This will reduce the use of conventional antibiotics, leading to increased antimicrobial resistance (Bondad-Reantaso et al.
, 2.
Streptococcus agalactiae.
Bacillus subtilis, and Staphylococcus aureus are aquatic bacterial pathogens that significantly impact the ecosystem.
These pathogens can be found in a variety of fish species and shrimp.
They persist in both freshwater and marine ecosystems, potentially leading to infection outbreaks (Wang et al.
, 2020.
Zelellw et al.
, 2021.
Chen et al.
, 2.
The present study focused on investigation of the antimicrobial activity of C.
asiatica leaves bioactive compounds on pathogenic bacteria by in vitro and in silico approach.
This study offers a novel approach to address antibiotic resistance in fish pathogenic bacteria, including S.
aureus and B.
The use of in silico methods to assess the affinity of these compounds for DNA gyrase offers important insights into their potential as antimicrobial agents.
This study investigates antibacterial activity by in vitro and continued with molecular docking of bioactive compounds from C.
asiatica, as potential inhibitors of DNA gyrase in pathogenic bacteria which is still rarely practiced today, and only a few data have been published.
Materials and Methods 1 Materials 1 The equipments The main equipment and tools used in this research included: vacuum rotary evaporator (Buchi.
Swis.
, spectrophotometer (Thermoscientific.
USA), bacterial incubator (Memmert.
German.
, micropipettes (Eppendorf.
German.
, microtips (Axygen.
USA), microtubes (Axygen.
USA), laboratory glassware (Pyrex.
USA).
Separating funnel (Schott Duran.
German.
Petridish (SPL Life Sciences.
South Kore.
, and 96well plate (Biologix.
USA).
2 The materials The plant material from Centella asiatica was obtained in Tegal Waru.
Ciampea.
Bogor Regency.
West Java.
Indonesia .
A34A19AS 106A41A58AE).
The leaves used in the study were old leaves.
Other materials used in this study were distilled water, ethanol (Merck.
USA), n-hexane (Merck.
USA), tryptic soy agar (Merck.
USA), mueller hinton broth (MHB) (Himedia.
Indi.
NaCl (Oxoid.
United Kingdo.
, and Phosphat Buffer Saline (Himedia.
Indi.
3 Ethical approval Ethical approval was not required for this study as no experimental animals were involved.
2 Methods 1 Identification of bioactive compounds The extraction procedure involved submerging 300 g of dried plant material in 1500 mL of a 70% ethanol solution.
The precipitate was then separated from the filtrate.
The filtrate was further concentrated with a rotary evaporator set at a temperature JIPK: Scientific Journal of Fisheries and Marine JIPK Vol 17 No 3.
October 2025 | In vitro and In silico Antibacterial Activity of Centella asiatica Leaves Bioactive.
between 40-45AC until a concentrated extract was obtained (Biradar and Rachetti, 2.
The GC-MS method was employed for the qualitative and quantitative characterization of C.
asiatica leaves extract (Magaya et al.
, 2.
Bioactive compound characterization was conducted in the Integrated Advanced Chemistry Laboratory.
Serpong-BRIN, using Gas Chromatography-Mass Spectrometry (GC-MS).
mg/mL in MHB medium.
A 96-microwell plate was utilized, where test tubes were filled with 160 AAL of MHB.
Subsequently, 20 AAL of crude extracts and fraction solutions at varying concentrations were added, followed by inoculation with 60 AAL of bacterial isolates at a density of 108 cfu/mL.
The mixture was then incubated for 24 hours.
The concentration of crude extracts that most effectively inhibits the Figure 1.
The minimum inhibitory concentration assay.
Agar well diffusion.
agalactiae isolate.
substilis isolate.
aureus isolate.
2 In vitro analysis Pathogenic bacterial isolates Bacillus subtilis (Inacc B1.
and Staphylococcus aureus (Inacc B.
were obtained from the Indonesian Culture Collection Laboratory (InaCC).
Cibinong.
BRIN.
Meanwhile.
Streptococcus agalactiae was isolated from infected fish.
In vitro crude extracts antibacterial activity was evaluated using modified minimum inhibitory concentration (MIC) methods.
The MIC was determined using the serial two-fold dilution method (Choudhury et al.
, 2.
Experiments involved preparing a solution of crude extracts .
at concentrations of 50, 25, 12.
5, 6.
25, 3.
563, 0.
781, 0.
390, 0.
195, 0.
098, 0.
049, and 0.
growth of selected bacteria is indicated by the precise visual assessment of the turbidity of the test Bacterial growth was observed visually, and the MIC value was established as the lowest concentration capable of halting bacterial growth, marked by a transition in color from yellow to pink.
In the MIC test.
Oxytetracycline antibiotic was used as the 3 In silico analysis The PubChem database was used to obtain the details of the bioactive compounds, including their LipinskiAos rules of five (Ro.
and ADME/T (Absorption.
Distribution.
Metabolism.
Excretion.
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JIPK: Scientific Journal of Fisheries and Marine JIPK Vol 17 No 3.
October 2025 | In vitro and In silico Antibacterial Activity of Centella asiatica Leaves Bioactive.
Figure 2.
3D and 2D interactions of four potential ligands and ciprofloxacin at the DNA gyrase site residues during molecular docking.
13-Hexyloxacyclotridec-10-en-2-one.
-muurolene.
and Toxicit.
In silico method applying molecular docking, alongside the assessment of Ro5 and ADME/T properties.
By analyzing the chemical properties of compounds.
LipinskiAos Ro5 serves as a method to predict their oral bioavailability.
LipinskiAos criteria suggest that a compound is more likely to be an effective oral drug if it possesses the following characteristics: .
a molecular weight (MW) not exceeding 500 Da, .
a partition coefficient .
ogP) less than or equal to 5, .
no more than 5 hydrogen bond donors (HBD), and .
no more than 10 hydrogen bond acceptors (HBA) (Frau et al.
Kumari and Kumar, 2.
The median lethal dosage (LD.
values were determined through an assessment of the toxicity class utilizing ProTox-II.
The in silico analysis material used three-dimensional structures of all ligands of bioactive compounds derived in .
sdf file format from the reputable National Center for Biotechnology Informa- tion (NCBI) PubChem database .
ttps://pubchem.
gov/).
At CB-Dock .
ttps://cadd.
cn/cb-dock2/index.
, molecular docking studies were conducted.
This involved identifying binding sites, assessing their dimensions and central coordinates, and adjusting the docking box size based on the ligands in the query.
AutoDock Vina performed molecular docking based on three-dimensional structures of specific proteins and analyzed the mechanisms of action of bioactive compounds (Eberhardt et al.
, 2021.
Xu et al.
, 2.
The proteins analyzed comprised DNA gyrase from S.
aureus (PDB ID: 6tc.
We assessed the typical inhibitor orientation in crystal structures.
The highly credible Protein Data Bank was referenced for the protein X-ray structures.
Binding postures and interaction diagrams were generated using BIOVIA Discovery Studio Visualizer 24.
ttps://w.
com/products/biovia/discovery-studio/visualizatio.
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Table 1.
Bioactive compounds identified in the leaves extract of Centella asiatica and their Lipinski properties No.
Compound Name
Area % HBA
HBD
TPSA
Log P .
LogP) LD50 Toxicity Class Terpenes Copaene Caryophyllene -Farnesene Humulene .
R,9R,E)-4,11,11-Trimethyl-8-methylenebicyclo.
undec-4-ene -Muurolene Caryophyllene oxide 2-Pentadecanone, 6,10,14-trimethyl- Phytol Fatty Acids Heptanal Hexanoic acid Heptanoic acid Hexadecanoic acid, methyl n-Hexadecanoic acid Ethyl 9-tetradecenoate Hexadecanoic acid, ethyl 9,12-Octadecadienoic acid, methyl ester JIPK: Scientific Journal of Fisheries and Marine JIPK Vol 17 No 3.
October 2025 | In vitro and In silico Antibacterial Activity of Centella asiatica Leaves Bioactive.
No.
Compound Name
Area % HBA
HBD
TPSA
Log P
LogP) LD50
Toxicity Class
9-Octadecenoic acid (Z)-, methyl ester
9,12-Octadecadienoic acid
(Z,Z)-
9,12,15-Octadecatrienoic acid, (Z,Z,Z)- Linoleic acid ethyl ester 9,12,15-Octadecatrienoic acid, ethyl ester, (Z,Z,Z)- Ricinoleic Acids Ricinoleic acid 9-Octadecenoic acid, 12-hydroxy-, methyl ester, [R-(Z)]- Acyclic Acids 2-Propenamide Ketones 13-Hexyloxacyclotridec-10-en-2-one Amines Benzyl alcohol, p-hydroxy-.
-[.
- Nitrosamines Ethanamine.
N-ethyl-N-nitroso- Pyrones 4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl- Copyright A2025 Faculty of Fisheries and Marine Universitas Airlangga Andriyanto et al.
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Compound Name
Area % HBA
HBD
TPSA
Log P .
LogP) LD50 Toxicity Class 2.
H)-Benzofuranone, 5,6,7,7a-tetrahydro-4,4,7a-trimethyl- No.
Lactones Alkenes Neophytadiene Benzofurans Loliolide Aldehydes Benzeneacetaldehyde Carbohydrates Erythritol Monosaccharide dl-Threitol Epoxide .
R,3E,7E,11R)-1,5,5,8-Tetramethyl-12-oxabicyclo.
dodeca-3,7-diene Others -D-Mannopyranoside, methyl 3,6-anhydro- Acetic acid, hydroxy-, ethyl Oxime-, methoxy-phenyl-_ Tetraacetyl-d-xylonic nitrile JIPK: Scientific Journal of Fisheries and Marine JIPK Vol 17 No 3.
October 2025 | In vitro and In silico Antibacterial Activity of Centella asiatica Leaves Bioactive.
No.
Compound Name
Area % HBA
HBD
TPSA
Log P .
LogP) LD50 Toxicity Class 2,4-Hexanedione, 5,5-dimethyl-1-phenyl- -Alanine.
TMS derivative Hydrazinecarboximidothioic acid, ethyl ester 11,11-Dimethyl-4,8-dimethylenebicyclo.
undecan-3-ol 2-Propenoic acid, pentadecyl -Methyl-3,4-.
phenethylamine Oxazepam, 2TMS derivative Decane, 3,8-dimethyl- 1-Octadecanesulphonyl chloride 2-.
Ao,4Ao,4Ao,6Ao,6Ao,8Ao,8Ao-Heptamethyltetrasiloxan- 2Ao-ylox .
-2,4,4,6,6,8,8,10,10-nonam Palmitic Acid.
TMS derivative Hexasiloxane, tetradecamethyl- Octadecanoic acid, 2-methyl-, methyl ester Description: MW .
olecular weight-g/mo.
HBA .
ydrogen bond acceptor.
HBD .
ydrogen bond donor.
TPSA .
opological polar surface are.
LD50 .
ethal dose-mg/k.
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Table 2.
Molecular docking results analysis of potential bioactive compounds identified from C.
asiatica and ciprofloxacin .
rug standar.
against DNA Gyrase (PDB id: 6tc.
DNA Gyrase No.
Compound Name Compound ID Binding Energy Cavity Volume 13-Hexyloxacyclotridec-10-en-2-one Oe7.
-Muurolene Oe6.
Copaene Oe6.
Humulene Oe6.
Caryophyllene Oe6.
9,12,15-Octadecatrienoic acid, ethyl ester, (Z,Z,Z)- Oe6.
2,4-Hexanedione, 5,5-dimethyl-1-phenyl- Oe6.
-Farnesene Oe6.
1-Octadecanesulphonyl chloride Oe6.
Phytol Oe6.
9,12,15-Octadecatrienoic acid, (Z,Z,Z)- Oe6.
R,9R,E)-4,11,11-Trimethyl-8-methylenebicyclo.
undec-4-ene Oe6.
Caryophyllene oxide Oe5.
Benzyl alcohol, p-hydroxy-.
-[.
Oe5.
R,3E,7E,11R)-1,5,5,8-Tetramethyl-12-oxabicyclo.
dodeca-3,7-diene Oe5.
9,12-Octadecadienoic acid, methyl ester Oe5.
JIPK: Scientific Journal of Fisheries and Marine JIPK Vol 17 No 3.
October 2025 | In vitro and In silico Antibacterial Activity of Centella asiatica Leaves Bioactive.
DNA Gyrase No.
Compound Name Compound ID Binding Energy Cavity Volume 2-Pentadecanone, 6,10,14-trimethyl- Oe5.
9,12-Octadecadienoic acid (Z,Z)- Oe5.
Oxime-, methoxy-phenyl-_ Oe5.
Hexadecanoic acid, ethyl ester Oe5.
9-Octadecenoic acid, 12-hydroxy-, methyl ester, [R-(Z)]- Oe5.
Ethyl 9-tetradecenoate Oe5.
2-Propenoic acid, pentadecyl ester Oe5.
Hexadecanoic acid, methyl ester Oe5.
Linoleic acid ethyl ester Oe5.
9-Octadecenoic acid (Z)-, methyl ester Oe4.
Erythritol Oe4.
dl-Threitol Oe4.
Heptanal Oe4.
Acetic acid, hydroxy-, ethyl ester Oe4.
-Alanine.
TMS derivative Oe4.
Oe7.
Ciprofloxacin .
rug standar.
Results and Discussion 1 Results 1 Gas chromatography analysis The GC-MS results of the C.
asiatica leaves extracts yielded 53 bioactive compounds (Table .
The compounds of C.
asiatica leaves extract include terpenes, fatty acids, ricinoleic acids, acyclic acids, ketones, amines, nitrosamines, pyrones, lactones, alkenes, benzofurans, aldehydes, carbohydrates, monosaccharides, epoxides, and various other component classes.
2 Antibacterial activity of C.
In vitro analysis shows that the minimum inhibitory concentration is the lowest concentration of antimicrobial agents capable of inhibiting the growth of harmful microorganisms.
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The MIC of crude extract was evaluated using the agar well diffusion assay (Figure .
The MIC values of C.
asiatica leaves extracts against S.
subtilis, and S.
aureus were 12.
5 mg/mL.
The results showed a positive antibacterial effect of C.
asiatica extract against Gram-positive bacteria.
In silico Ro5 and ADME/T analysis identified 31 potential drug-candidate compounds with antibacterial properties (Table .
The 31 active compounds derived from the leaves extract of C.
asiatica demonstrated molecular weights between 1 Da and 353 Da.
The toxicity assessment of 31 compounds indicated an LD50 toxicity range of 2,000 to 34,900 mg/kg.
Figure 2 illustrates the comparison of binding patterns and molecular interactions of the evaluated compounds with the highest binding energies against the drug standard ciprofloxacin, recognized as a DNA gyrase inhibitor.
The four highest-ranking ligands for DNA Gyrase, determined by Vina score, are 13-Hexyloxacyclotridec-10-en-2-one (Oe7.
4 kcal/ mo.
, -Muurolene (Oe6.
7 kcal/mo.
Copaene (Oe6.
kcal/mo.
, and Humulene (Oe6.
6 kcal/mo.
2 Discussion 1 Gas chromatography identification GC-MS analysis of C.
asiatica leaves extract in accordance with previous studies by Micheli et al.
Jenitha .
, and Taleghani et al.
found the metabolites of C.
asiatica, which include triterpenoids, phenolics, flavonoids, phenylpropanoids, acyclic acids, ketones, and amines.
The findings align with those of Yang et al.
Pillai et al.
, and Rafi et al.
, which indicate that C.
asiatica comprises numerous bioactive components, such as terpenoids, flavonoids, saponins, tannins, amino acids, fatty acids, alkaloids, steroids, and and other categories.
According to Sieberi et al.
and Taghizadeh and Jalili .
that C.
asiatica bioactive compounds, especially triterpenoids, flavonoids, and phenolic compounds, play an important role as antibacterial agents.
Likewise.
Pham et al.
and Menon et al.
mentioned that the antibacterial properties of phytochemical compounds contained in C.
asiatica are applied to a variety of pathogenic microbial organisms.
2 In vitro and in silico antibacterial activity The antibacterial activity of C.
leaves extracts against three pathogenic bacteria (S.
subtilis, and S.
was determined.
In the antibacterial activity, the extract con- centration is significant in preventing the growth of pathogenic bacteria.
The MIC values against S.
subtilis and S.
aureus were 12.
5 mg/mL.
The results are in accordance with those reported by Zhang et al.
Qurrotuaini et al.
, and Kathirvel et al.
asiatica leaves extracts exhibit activity against Acinetobacter calcoaceticus anitratus.
Bacillus cereus.
Enterococcus avium.
Klebsiella pneumoniae.
Proteus mirabilis.
Pseudomonas aeruginosa.
Salmonella typhi.
Staphylococcus aureus, and Streptococcus agalactiae with a MIC ranging from 1.
25 to 25 mg/mL.
The results indicate that C.
asiatica has a significant inhibitory effect on the growth of Gram-positive bacteria within 24 hours.
Sieberi et .
reported that ethanol and Dichloro methane (DCM) extracts of C.
asiatica inhibit the growth of Gram-positive and Gram-negative bacteria.
While Menon et al.
suggested that in vitro studies have also shown a significant reduction in the number of colonies of pathogenic bacteria after treatment with C.
asiatica extract.
The antibacterial mechanism of bioactive compounds in C.
asiatica functions synergistically within bacterial cells by inhibiting nucleic acid synthesis, which is thought to involve the loss of bacterial membrane integrity.
This results in increased permeability and subsequent cell death and influences the bacterial metabolic system (Wong and Ramli, 2021.
Maitra et al.
, 2022.
Qurrotuaini et al.
Wei et al.
, 2.
Another study found that the antibacterial mechanism is related to the inhibition of quorum-sensing activity that prevents communication between bacteria in biofilm formation as well as in increased pathogenicity (Sieberi et al.
, 2020.
Taghizadeh and Jalili, 2.
The results of the in silico analysis showed that 31 bioactive compounds passed the Lipinski Ro5 and ADME/T test.
Zafar et al.
and Nguyen et al.
reported that Lipinski Ro5 indicate that a molecular weight of under 500 Da implies potential for cellular membrane penetration.
Both compounds exhibited HBA.
HBD, and iLog P values of less than 10, less than 5, and less than 5, respectively, while the TPSA value was less than or equal to 140 yI.
LD50 value indicates reduced chemical toxicity to the tested organism.
Determining toxicity levels using computer-based tools such as ProTox-II and Swiss ADME in molecular docking can facilitate the classification of bioactive compounds based on their toxicity in accordance with standard drug criteria (Lane et al.
, 2023.
Li et al.
, 2024.
Ghanem et al.
, 2.
Abishad et al.
and Wu et al.
suggested that enhancing the safety of these JIPK: Scientific Journal of Fisheries and Marine JIPK Vol 17 No 3.
October 2025 | In vitro and In silico Antibacterial Activity of Centella asiatica Leaves Bioactive.
drugs prior to market introduction requires a focus on ADME and toxicity-related factors.
The SWISS ADME online program was employed to assess the drug-likeness of phytocompounds.
The molecular factors associated with rule violations and the acquisition of bioactive compounds are detailed in the table of Lipinski parameters.
The results of in vitro studies demonstrated the antibacterial potential of C.
asiatica leaves extracts, so further in silico evaluations were carried out to identify the compounds that could significantly inhibit DNA gyrase, a critical enzyme in bacterial cell development.
In the docking investigations of the DNA gyrase binding site, 13-Hexyloxacyclotridec-10-en-2-one had the greatest binding energy of Oe7.
4 kcal/mol surpassing drug standard ciprofloxacin with binding energy Oe7.
3 kcal/mol.
Based on previous research.
Selvarajan et al.
reported that 13OeHexyloxacyclotridecOe10OeenOe2Oeone exhibits wide antibacterial activity against numerous pathogenic bacteria.
Singh et al.
also stated that 13OeHexyloxacyclotridecOe10OeenOe2Oeone efficiently suppresses the development of bacteria such as Staphylococcus aureus and Escherichia coli.
a broad-spectrum antibiotic in the fluoroquinolone group, ciprofloxacin is widely utilized to treat a range of bacterial infections, including those caused by both Gram-positive and Gram-negative bacteria.
Research reveals that ciprofloxacin exerts its bactericidal action by binding to the bacterial enzymes DNA gyrase and topoisomerase IV, which prevents DNA replication from occurring (Hussein et al.
GrigorAoeva et al.
, 2.
Furthermore, the approach illustrated how C.
asiatica demonstrates its antibacterial activity by disrupting bacterial DNA The bioactive compounds of C.
can elicit apoptosis through mechanisms involving DNA synthesis (Jenitha, 2.
This observation underscores a crucial connection to the role of DNA gyrase, a crucial enzyme in bacterial DNA replication.
Inhibition of DNA gyrase obstructs bacterial replication, hence averting infection.
Moreover, studies reveal that C.
asiatica extracts, with their considerable antibacterial activity at minimal concentrations, hold great potential as agents that inhibit bacterial proliferation (Agneeswari et al.
, 2.
It appeared beneficial for performing molecular docking studies that align in silico and in vitro.
results depending on the findings of the in vitro inquiry.
This study employed molecular docking analysis via the CB-Dock server and GC-MS analysis to evaluate the interactions between the bioactive compounds in C.
asiatica leaves extracts and the target protein DNA gyrase.
Eberhardt et al.
and Wang et al.
suggested that docking studies are employed in drug development to forecast the interactions between ligands and receptors, as well as to rank compounds according to binding energies or fitness scores.
While Liu et al.
and Zheng et al.
stated that the CB-Dock methodology consists of three phases: first, assessing the curvature of the protein surface.
second, clustering to pinpoint active site cavities.
and third, performing docking with AutoDock Vina.
The rise of bacterial resistance to existing treatment agents has prompted the development of new antimicrobial drugs aimed at selectively inhibiting evolving bacterial targets that face ongoing This study demonstrates that molecular docking analysis shows 13OeHexyloxacyclotridecOe10OeenOe2Oeone possesses greater selectivity for the DNA gyrase binding site than ciprofloxacin, the standard medication.
The compound 13-Hexyloxacyclotridec-10-en-2-one may offer a solid starting point for developing novel chemical entities that exhibit potent antibacterial effects.
The results suggest that bioactive compounds derived from C.
leaves extracts could function as effective antibacterial agents against fish pathogenic bacteria.
Conclusion The bioactive compounds of Centella asiatica leaves extracts were analyzed via GC-MS, encompassing terpenes, fatty acids, ricinoleic acids, acyclic acids, ketones, amines, nitrosamines, pyrones, lactones, alkenes, benzofurans, aldehydes, carbohydrates, monosaccharides, epoxides, and other compounds classes.
The leaves extract demonstrated antibacterial effectiveness against fish pathogenic bacteria (Streptococcus agalactiae.
Bacillus subtilis, and Staphylococcus aureu.
with MIC values of 12.
5 mg/mL.
Through in silico analysis, 31 compounds met the criteria of five drug-likeness features.
Furthermore, molecular docking investigations showed that 13-Hexyloxacyclotridec-10-en-2-one had the most antibacterial activity.
The results demonstrated that bioactive compounds from Centella asiatica leaves extracts have the potential as antibacterial agents.
Acknowledgement The authors would like to thank Research Centre of Veterinary Science, the Research Organization for Health-BRIN for support this Copyright A2025 Faculty of Fisheries and Marine Universitas Airlangga Andriyanto et al.
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AuthorsAo Contributions All authors have contributed to the final Each authorAos contribution is as follows.
SA.
designed the experiment, collected data, and writing-original manuscript.
NN.
LG.
HN.
analysis and editing.
MM.
SA.
MN.
writing-review and critical revision of the article.
Conflict of Interest The authors declare that there is no conflict of interest.
Declaration of Artificial Intelligence (AI) The author.
affirm that no artificial intelligence (AI) tools, services, or technologies were employed in the creation, editing, or refinement of this All content presented is the result of the independent intellectual efforts of the author.
, ensuring originality and integrity.
Funding Information This research was supported by the Research Organization for Health-the National Research and Innovation Agency.
References