Coleus amboinicus (Lour.) is a medicinal plant containing various bioactive compounds. Endophytes are microorganisms living inside intracellular tissue of plants and known as the source of bioactive compounds. In order to explore the potential of endophyte in producing novel bioactive compounds, this study focused on isolating endophytic fungi from the leaves of C.amboinicus, characterisation and screening their metabolite bioactivity during submerged culture fermentation. Isolation of endophytic fungi from the leaves segment of C.amboinicus was conducted on PDA media and fungus identification was carried out by analyzing its morphology and molecular examination. The production of metabolites was examined using thin layer chromatography and gas chromatography. Identification of the endophytic fungus showed 98.84% similarity with Eutypa linearis. This species was fermented submergedly in PDB medium for 14 days under dark and exposed to light and the fermentation broth was extracted using ethyl acetate. The results showed that exposure to the light did not significantly influence metabolite production. The ethyl acetate extract exhibited antioxidant and cytotoxic activities. Antioxidant activity of this extract as examined by DPPH assay showed IC50 of 105.31 ± 2.11 µg/mL. Cytotoxic activity against Hela cell line was known to be the best among other cell lines (IC50 301.53 ± 11.34 µg/mL) although it was found to be non selective (SI<1). The extract contained five major compounds namely Benzenemethanol, 4-nitro-(CAS) p-Nitrobenzyl alcohol; 2-Pentadecanone (CAS) Pentadecan-2-one; (1R*,6S*,10R*)-5,5-Dimethyl-11,12-dioxatricyclo[8.2.1.0(1,6)] tridecan-10-ol; 9,12-Octadecadienoic acid (Z,Z)-, methyl ester (CAS) Methyl linoleate; and 3-Furanacetic acid, 4-hexyl-2,5-dihydro-2,5-dioxo- (CAS) 2-carboxymethyl-3-N-hexyl-maleic anhydride.
Abstract. Eltivitasari A, Rahmawati, Gemantari BM, Romadhonsyah F, Nurrochmad A, Wahyuono S, Astuti P. 2021. Effect of light exposure on secondary metabolites production of an endophytic fungus Arthrinium rasikravindrae and its antioxidant and anticancer activities. Biodiversitas 22: 3156-3163. Endophytic microorganisms are one of the promising sources in producing bioactive compounds, to be developed for new drug candidates. They are found to have the ability to generate the same compounds as their host plant. Metabolite producing capacity of the endophytes is known to be affected by light exposure during fermentation process. This study focused on an endophytic fungus Arthrinium rasikravindrae isolated from Coleus amboinicus stem to reveal out its metabolite profiles due to light exposure as well as its bioactivity consequences. A. rasikravindrae was cultured on potato dextrose broth medium for 14 days and fermented in dark and exposed to natural light. Metabolite profiling was performed using TLC and GC-MS analysis. The activities were observed using DPPH assay for antioxidant and MTT assay for cytotoxicity potential. The results showed that A. rasikravindrae ethyl acetate extract produced during dark and exposed to light fermentation conditions contained different compounds but there was some which showed similarity with their host plant. Methyl octadec-9-enoate was found in all fermentation conditions as well as in C. amboinicus stem extract. Besides methyl octadec-9-enoate, methyl palmitate was also found present in both A. rasikravindrae extract fermented exposed to light and its host plant. The antioxidant activity of extract generated from dark fermentation condition was better as compared to that exposed to light with IC50 value of 66.36±0.53 vs 556.92±34.37 µg/mL. However, cytotoxic activity screening against several cancer cell lines exhibited opposing results in which extract from light-exposed fermentation resulted in better cytotoxic activity (IC50 value of 291.40 ± 2.34 µg/mL on WiDr, 336.80 ± 5.05 µg/mL on T47D, and 404.73 ± 3.46 µg/mL on Hela cell lines). Extract obtained from dark fermentation condition showed IC50 value of more than 500 µg/mL in all tested cancer cell lines. Preliminary examination on cytotoxic activity against WiDR cells suggested that the extract from light-exposed fermentation might induce cell death through mechanisms involving cell cycle arrest.
Fungi endofit banyak dilaporkan mampu memproduksi senyawa metabolit yang memiliki beragam aktivitas biologis. Diisolasi dari salah satu tanaman berkhasiat Indonesia yang dikenal dengan nama tanaman jinten (Coleus amboinicus Lour.), pada penelitian ini dilakukan observasi aktivitas antimikroba fungi endofit Eutypa linearis. Ekstrak etil asetat dari media fermentasi E.linearis dalam beragam konsentrasi diujikan kepada beberapa mikroba bakteri dan fungi yakni Staphyloccocus aureus, Pseudomonas aeruginosa, Bacillus subtilis, dan Candida albicans. Aktivitas antimikroba dievaluasi menggunakan metode mikrodilusi pada media Mueller Hinton Broth untuk kultur bakteri dan Brain Heart Infussion Broth untuk kultur fungi. Nilai optical density pada panjang gelombang 600 nm yang diperoleh dari penggunaan microplate reader digunakan untuk menghitung nilai IC50 ekstrak. Hasil penelitian menunjukkan nilai IC50 ekstrak etil asetat media fermentasi E.linearis pada S.aureus adalah sebesar 260,69 µg/mL; P.aeruginosa 359,32 µg/mL; B.subtilis 163,74 µg/mL; dan C.albicans 92,75 µg/mL. Berdasarkan metode bioautografi kontak, diketahui senyawa aktif sebagai antimikroba tersebut merupakan kelompok senyawa yang polar.
The aim of this study was to determine the physical and chemical stabilities of tablets containing Sonchus arvensis L leaves extract and Lumbricus rubellus. The tablets were produced by wet granulation process. The stability test was observed during a 3-month accelerated stability under 40 ± 2ºC and 75 ± 5% RH storage conditions. The tablets were evaluated every month for the physical properties include organoleptic, weight variation, friability, hardness, and disintegration time, while the chemical properties of the tablets include Rf value, and spotting profile was examined using TLC-Densitometry. The result of the physical stability test up to the third month showed that the tablets of Sonchus arvensis leaves extract and Lumbricus rubellus met the requirements of Indonesian Pharmacopeia 5th and United State Pharmacopeia 32. The qualitative chemical property stability test on the tablets also demonstrated that luteolin as a marker compound remained stable in the tablets after three months of storage in an accelerated condition. Conclusion, the formulation of a tablet containing Sonchus arvensis leaves extract and Lumbricus rubellus stable after this stability studies.
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