Antibacterial Effect of an Ethyl Acetate Extract of Emericella nidulans Derived from Endophytic Fungus Rhizophora mucronata Against Methicillin-Resistant Staphylococcus aureus
DOI:
https://doi.org/10.48048/tis.2025.9130Keywords:
Emericella nidulans, MRSA, Molecular docking, PBP2a, Bacterial resistance, Rhizopora mucronataAbstract
Methicillin-resistant Staphylococcus aureus (MRSA) poses a significant global public health challenge due to its resistance to conventional antibiotics, leading to persistent infections, increased healthcare costs, and high mortality rates. MRSA can cause various diseases, including those affecting the skin, subcutaneous tissues, and internal organs. Its resistance is largely driven by the mecA gene, which encodes Penicillin Binding Protein 2a (PBP2a), which prevents β-lactam antibiotics from effectively targeting the bacterial cell wall. In this study, crude extract and fractions of Emericella nidulans derived from Rhizophora mucronata mangrove were prepared and screened for antibacterial activity against MRSA. In vitro tests on ethyl acetate extracts and fractions of E. nidulans revealed that they can inhibit MRSA growth in a moderate range. The crude extracts were analyzed using liquid chromatography-high-resolution mass spectrometry (LC-HRMS) to determine the compound contained. Forty-eight compounds were screened through molecular docking analysis to identify the bioactive compounds inhibiting the SauPBP2a receptor. It is indicated that E. nidulans shows promise as raw materials for antibacterial against MRSA.
HIGHLIGHTS
- Ethyl acetate extract of nidulans showed antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) in vitro and contained 47 compounds based on LC-HRMS analysis.
- In silico studies predicted nine compounds such as 1-linoleoyl glycerol, 9-oxo-10(E),12(E)-octadecadienoic acid, monoolein, methyl ester of 9(Z),11(E),13(E)-octadecadienoic acid, bis(4-ethylbenzylidene) sorbitol, α-linolenic acid, and oleamide have ability to inhibit the SauPBP2a receptor.
GRAPHICAL ABSTRACT

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P Nandhini, PK Gupta, AK Mahapatra, AP Das, SM Agarwal, S Mickymaray, A Alothaim and M Rajan. In-silico molecular screening of natural compounds as a potential therapeutic inhibitor for Methicillin-resistant Staphylococcus aureus inhibition. Chemico-Biological Interactions 2023; 374, 110383.
PY Ong. Recurrent MRSA skin infections in atopic dermatitis. The Journal of Allergy and Clinical Immunology: In Practice 2014; 2(4), 396-399.
KL Urish and JE Cassat. Staphylococcus aureus osteomyelitis: Bone, bugs, and surgery. Infection and Immunity 2020; 88(7), e00932-19.
WD Longhurst and JM Sheele. Spontaneous methicillin-resistant Staphylococcus aureus (MRSA) meningitis. The American Journal of Emergency Medicine 2018; 36(5), 909.
KZ Vardakas, DK Matthaiou and ME Falagas. Incidence, characteristics and outcomes of patients with severe community acquired-MRSA pneumonia. European Respiratory Journal 2009; 34(5), 1148-1158.
AD Lewis, MR Bridwell, MD Hambuchen, TB Clay and KW Orwig. Correlation of MRSA polymerase chain reaction (PCR) nasal swab in ventilator-associated pneumonia, lung abscess, and empyema. Diagnostic Microbiology and Infectious Disease 2023; 105(1), 115836.
SP Bergin, TL Holland, VG Fowler and SYC Tong. Bacteremia, sepsis, and infective endocarditis associated with Staphylococcus aureus. Current Topics in Microbiology and Immunology 2017; 409, 263-296.
R Tabassum, S Kousar, G Mustafa, A Jamil and SA Attique. In silico method for the screening of phytochemicals against methicillin‐resistant Staphylococcus aureus. BioMed Research International 2023; 2023(1), 5100400.
M Fitranda, SIO Salasia, O Sianipar, DA Dewananda, AZ Arjana, F Aziz, M Wasissa, FB Lestari and CM Santosa. Methicillin-resistant Staphylococcus aureus isolates derived from humans and animals in Yogyakarta, Indonesia. Veterinary World 2023; 16(1), 239-245.
Y Katayama, HZ Zhang and HF Chambers. PBP 2a mutations producing very-high-level resistance to beta-lactams. Antimicrobial Agents and Chemotherapy 2004; 48(2), 453-459.
JM Intravia, MN Osterman and R Tosti. Antibiotic management and antibiotic resistance in hand infections. Hand Clinics 2020; 36(3), 301-305.
R Sobral and A Tomasz. The staphylococcal cell wall. Microbiology Spectrum 2019. http://doi.org/10.1128/microbiolspec.gpp3-0068-2019.
D Lim and NCJ Strynadka. Structural basis for the β lactam resistance of PBP2a from methicillin-resistant Staphylococcus aureus. Nature Structural Biology 2002; 9(11), 870-876.
J Zhou, Z Feng, W Zhang and J Xu. Evaluation of the antimicrobial and cytotoxic potential of endophytic fungi extracts from mangrove plants Rhizophora stylosa and R. mucronata. Scientific Reports 2022; 12(1), 2733.
KF Ghazawi, SA Fatani, SGA Mohamed, GA Mohamed and SRM Ibrahim. Aspergillus nidulans-Natural metabolites powerhouse: Structures, biosynthesis, bioactivities, and biotechnological potential. Fermentation 2023; 9(4), 325.
J Kjer, A Debbab, AH Aly and P Proksch. Methods for isolation of marine-derived endophytic fungi and their bioactive secondary products. Nature Protocols 2010; 5(3), 479-490.
A Windarsih, Suratno, HD Warmiko, AW Indrianingsih, A Rohman and YI Ulumuddin. Untargeted metabolomics and proteomics approach using liquid chromatography-Orbitrap high resolution mass spectrometry to detect pork adulteration in Pangasius hypopthalmus meat. Food Chemistry 2022; 386, 132856.
M Balouiri, M Sadiki and SK Ibnsouda. Methods for in vitro evaluating antimicrobial activity: A review. Journal of Pharmaceutical Analysis 2016; 6(2), 71-79.
S Abraham, A Basukriadi, S Pawiroharsono and W Sjamsuridzal. Insecticidal activity of ethyl acetate extracts from culture filtrates of mangrove fungal endophytes. Mycobiology 2015; 43(2), 137-149.
A Maurya, K Kalani, SC Verma, R Singh and A Srivastava. Vacuum liquid chromatography: Simple, efficient and versatile separation technique for natural products. Organic & Medicinal Chemistry International Journal 2018; 7(2), 80-82.
R Tautenhahn, C Böttcher and S Neumann. Highly sensitive feature detection for high resolution LC/MS. BMC Bioinformatics 2008; 9, 504.
E Chekmeneva, GDS Correia, M Gómez-Romero, J Stamler, Q Chan, P Elliott, JK Nicholson and E Holmes. Ultra-performance liquid chromatography-high-resolution mass spectrometry and direct infusion-high-resolution mass spectrometry for combined exploratory and targeted metabolic profiling of human urine. Journal of Proteome Research 2018; 17(10), 3492-3502.
S Singha, V Gowda and MS Hedenqvist. Plant cuticle-inspired polyesters as promising green and sustainable polymer materials. ACS Applied Polymer Materials 2021; 3(8), 4088-4100.
Y Wang, D Li, J Gao, X Li, R Zhang, X Jin, Z Hu, B Zheng, S Persson and P Chen. The 2'-O-methyladenosine nucleoside modification gene OsTRM13 positively regulates salt stress tolerance in rice. Journal of Experimental Botany 2017; 68(7), 1479-1491.
J Das, R Kumar, SK Yadav and G Jha. Nicotinic acid catabolism modulates bacterial mycophagy in Burkholderia gladioli strain NGJ1. Microbiology Spectrum 2023; 11(3), e0445722.
N Fujimori and H Ashihara. Adenine metabolism and the synthesis of purine alkaloids in flowers of Camellia. Phytochemistry 1990; 29(11), 3513-3516.
A Turk, S Lee, SW Yeon, SH Ryu, YK Han, YJ Kim, SM Ko, BS Kim, BY Hwang, KY Lee and MK Lee. Adenosine deaminase inhibitory activity of medicinal plants: Boost the production of cordycepin in cordyceps militaris. Antioxidants 2023; 12(6), 1260.
MK Arumugam, MC Paal, TMJ Donohue, M Ganesan, NA Osna and KK Kharbanda. Beneficial effects of betaine: A comprehensive review. Biology 2021; 10(6), 456.
D Dobrijević, K Pastor, N Nastić, F Özogul, J Krulj, B Kokić, E Bartkiene, JM Rocha and J Kojić. Betaine as a functional ingredient: Metabolism, health-promoting attributes, food sources, applications and analysis methods. Molecules 2023; 28(12), 4824.
W Wang, S Yamaguchi, M Koyama, S Tian, A Ino, K Miyatake and K Nakamura. LC-MS/MS analysis of choline compounds in Japanese-cultivated vegetables and fruits. Foods 2020; 9(8), 1029.
C Akin-Osanaiye, A Gabriel and R Alebiosu. Characterization and antimicrobial screening of ethyl oleate isolated from Phyllanthus Amarus (Schum and Thonn). Annuals of Biological Research 2011; 2(2), 298-305.
S Ramachandran, P Fontanille, A Pandey and C Larroche. Gluconic acid: Properties, applications and microbial production. Food Technology & Biotechnology 2006; 44(2), 185-195.
A Durazzo, M Lucarini, A Nazhand, SB Souto, AM Silva, P Severino, EB Souto and A Santini. The nutraceutical value of carnitine and its use in dietary supplements. Molecules 2020; 25(9), 2127.
ACMF Araújo, WMC Araújo, UML Marquez, R Akutsu and EY Nakano. Table of phenylalanine content of foods: Comparative analysis of data compiled in food composition tables. JIMD Reports 2017; 34, 87-96.
MR Hasan, MM Haque, MA Hoque, S Sultana, MM Rahman, MAA Shaikh and MKU Sarker. Antioxidant activity study and GC-MS profiling of Camellia sinensis Linn. Heliyon 2024; 10(1), e23514.
E Okuyama, T Hasegawa, T Matsushita, H Fujimoto, M Ishibashi and M Yamazaki. Analgesic components of saposhnikovia root (Saposhnikovia divaricata). Chemical & Pharmaceutical Bulletin 2001; 49(2), 154-160.
G Chen, M Zhu and M Guo. Research advances in traditional and modern use of Nelumbo nucifera: Phytochemicals, health promoting activities and beyond. Critical Reviews in Food Science and Nutrition 2019; 59(S1), S189-S209.
PW Harlina, V Maritha, R Shahzad, M Rafi, F Geng, I Musfiroh, M Muchtaridi, R Wahab, AA Al-Khedhairy, S Koerniati and NN Amalina. Comprehensive profiling and authentication of porcine, bovine, and goat bone gelatins through UHPLC-HRMS metabolomics and chemometric strategies. LWT 2024; 205, 116529.
CY Ma, WK Liu and CT Che. Lignanamides and nonalkaloidal components of Hyoscyamus niger seeds. Journal of Natural Products 2002; 65(2), 206-209.
D Wang, TJ Girard, TP Kasten, RM LaChance, MA Miller-Wideman and RC Durley. Inhibitory activity of unsaturated fatty acids and anacardic acids toward soluble tissue factor-factor VIIa complex. Journal of Natural Products 1998; 61(11), 1352-1355.
RM Munavu. Fatty acid composition of seed kernel oil of Calodendrum capense (L.f.) Thunb. Journal of the American Oil Chemists Society 1983; 60(9), 1653-1653.
K Machida, LS Trifonov, WA Ayer, ZX Lu, A Laroche, HC Huang, KJ Cheng and JL Zantige. 3(2H)-Benzofuranones and chromanes from liquid cultures of the mycoparasitic fungus Coniothyrium minitans. Phytochemistry 2001; 58(1), 173-177.
M Isaka, P Chinthanom, S Kongthong, S Supothina and P Ittiworapong. Hamigeromycins C-G, 14-membered macrolides from the fungus Hamigera avellanea BCC 17816. Tetrahedron 2010; 66(4), 955-961.
K Fidyt, A Fiedorowicz, L Strządała and A Szumny. β‐caryophyllene and β‐caryophyllene oxide-natural compounds of anticancer and analgesic properties. Cancer Medicine 2016; 5(10), 3007-3017.
C Delgado, G Mendez-Callejas and C Celis. Caryophyllene oxide, the active compound isolated from leaves of Hymenaea courbaril L.(Fabaceae) with antiproliferative and apoptotic effects on PC-3 androgen-independent prostate cancer cell line. Molecules 2021; 26(20), 6142.
X Liu and W Quan. Progress on the synthesis pathways and pharmacological effects of naturally occurring pyrazines. Molecules 2024; 29(15), 3597.
K Mogi, Y Koya, M Yoshihara, M Sugiyama, R Miki, E Miyamoto, H Fujimoto, K Kitami, S Iyoshi, S Tano, K Uno, S Tamauchi, A Yokoi, Y Shimizu, Y Ikeda, N Yoshikawa, K Niimi, Y Yamakita, H Tomita, K Shibata, A Nawa, Y Tomoda and H Kajiyama. 9-oxo-ODAs suppresses the proliferation of human cervical cancer cells through the inhibition of CDKs and HPV oncoproteins. Scientific Reports 2023; 13(1), 19208.
ZP Yu, C An, Y Yao, CY Wang, Z Sun, C Cui, L Liu and SS Gao. A combined strategy for the overproduction of complex ergot alkaloid agroclavine. Synthetic and Systems Biotechnology 2022; 7(4), 1126-1132.
MM Gutiérrez-Pacheco, H Torres-Moreno, ML Flores-Lopez, NV Guadarrama, JF Ayala-Zavala, LA Ortega-Ramírez and JC López-Romero. Mechanisms and applications of citral’s antimicrobial properties in food preservation and pharmaceuticals formulations. Antibiotics 2023; 12(11), 1608.
SA Ashraf, AEO Elkhalifa, AJ Siddiqui, M Patel, AM Awadelkareem, M Snoussi, MS Ashraf, M Adnan and S Hadi. Cordycepin for health and wellbeing: A potent bioactive metabolite of an entomopathogenic medicinal fungus Cordyceps with its nutraceutical and therapeutic potential. Molecules 2020; 25(12), 2735.
K Naumoska, U Jug, V Metličar and I Vovk. Oleamide, a bioactive compound, unwittingly introduced into the human body through some plastic food/beverages and medicine containers. Foods 2020; 9(5), 549.
KK Yadav, N Gupta, S Prasad, LC Malav, JK Bhutto, A Ahmad, A Gacem, BH Jeon, AM Fallatah, BH Asghar, MMS Cabral-Pinto, NS Awwad, OKR Alharbi, M Alam and S Chaiprapat. An eco-sustainable approach towards heavy metals remediation by mangroves from the coastal environment: A critical review. Marine Pollution Bulletin 2023; 188, 114569.
MUE Sanam, AIR Detha and NK Rohi. Detection of antibacterial activity of lactic acid bacteria, isolated from Sumba mare’s milk, against Bacillus cereus, Staphylococcus aureus, and Escherichia coli. Journal of Advanced Veterinary and Animal Research 2022; 9(1), 53-58.
V Jeyanthi and P Velusamy. Anti-methicillin resistant Staphylococcus aureus compound isolation from halophilic Bacillus amyloliquefaciens MHB1 and determination of its mode of action using electron microscope and flow cytometry analysis. Indian Journal of Microbiology 2016; 56(2), 148-157.
MAW Shalaby, EME Dokla, RAT Serya and KAM Abouzid. Penicillin binding protein 2a: An overview and a medicinal chemistry perspective. European Journal of Medicinal Chemistry 2020; 199, 112312.
B Morak-Młodawska, M Jeleń, E Martula and R Korlacki. Study of lipophilicity and ADME properties of 1,9-diazaphenothiazines with anticancer action. International Journal of Molecular Sciences 2023; 24(8), 6970.
D Gadaleta, K Vuković, C Toma, GJ Lavado, AL Karmaus, K Mansouri, NC Kleinstreuer, E Benfenati and A Roncaglioni. SAR and QSAR modeling of a large collection of LD50 rat acute oral toxicity data. Journal of Cheminformatics 2019; 11(1), 58.
J Raj, M Chandra, TD Dogra, M Pahuja and A Raina. Determination of median lethal dose of combination of endosulfan and cypermethrin in wistar rat. Toxicology International 2013; 20(1), 1-5.
M Masumi, F Noormohammadi, F Kianisaba, F Nouri, M Taheri and A Taherkhani. Methicillin‐resistant Staphylococcus aureus: Docking‐based virtual screening and molecular dynamics simulations to identify potential penicillin‐binding protein 2a inhibitors from natural flavonoids. International Journal of Microbiology 2022; 2022, 9130700.
SJ Uddin, J Bettadapura, P Guillon, ID Grice, S Mahalingam and E Tiralongo. In-vitro antiviral activity of a novel phthalic acid ester derivative isolated from the Bangladeshi mangrove fern Acrostichum aureum. Journal of Antivirals & Antiretrovirals 2013; 5(6), 139-144.
ALDS Pontes, VC Mesquita, FDO Chaves, AJRD Silva, MAC Kaplan and CE Fingolo. Phthalates in Avicennia schaueriana, a mangrove species, in the State Biological Reserve, Guaratiba, RJ, Brazil. Environmental Advances 2020; 2, 100015.
CP Victório, MSD Santos and NK Simas. Phthalates: Environmental pollutants detected in leaf epicuticular wax of Avicennia schaueriana and Rhizophora mangle from a mangrove ecosystem. International Journal of Environmental Studies 2022; 79(1), 114-123.
HJ Kwon, H Ahn, BS Kim, SS Kang and KG Lee. Anti-bacterial and anti-inflammatory activities of lactic acid bacteria-bioconversioned indica rice (Oryza sativa L.) extract. Chemical and Biological Technologies in Agriculture 2022; 9, 44.
SB Izzati, SIN Anidah and AA Yusmur. Antibacterial activity profile of mangrove endophytic fungi isolated from berau regency, Indonesia. Indonesian Journal of Biotechnology & Bioscience 2022; 9(1), 86-99.
D Kusumah, M Wakui, M Murakami, X Xie, K Yukihito and I Maeda. Linoleic acid, α-linolenic acid, and monolinolenins as antibacterial substances in the heat-processed soybean fermented with Rhizopus oligosporus. Bioscience, Biotechnology, and Biochemistry 2020; 84(6), 1285-1290.
BK Yoon, JA Jackman, ER Valle-González and NJ Cho. Antibacterial free fatty acids and monoglycerides: Biological activities, experimental testing, and therapeutic applications. International Journal of Molecular Sciences 2018; 19(4), 1114.
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