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All Journal Indonesian Journal of Biotechnology Majalah Obat Tradisional
Subagus Wahyuono
Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, Indonesia
Biochemistry, Genetics & Molecular Biology Health Professions Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology Public Health

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Identifcation of antibiotic producing endophytic microbe isolates from a national park in Java island Sri Yuwantiningsih; Sebastian Margino; Subagus Wahyuono
Indonesian Journal of Biotechnology Vol 20, No 2 (2015)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (786.998 KB) | DOI: 10.22146/ijbiotech.24345

Abstract

Endophytic microbes are potential sources of antibiotics. Some numbers of endophytic bacteria were isolated from plants in Ujung Kulon, Kaliurang, Meru Betiri and Baluran National Park, Bogor Botanical Garden, and Nusakambangan forest, Indonesia. Previous studies have been conducted to examine and obtain endophytic bacteria isolates from the selected plants, which resulted in three selected isolates, namely OOH-1, STG-1, and CMB-2. This research was conducted to determine the molecular identity of OOH-1 and STG-1 isolates, as well as to identify antibiotic compounds produced by STG-1 isolate. Molecular identifcation of selected isolates was based on 16S rRNA gene analysis and amplifed using primers 27F and 1492R. A phylogeny tree was then constructed by comparing the resulting sequences with data from Gene Bank using the BLAST-N program. The identifcation showed that STG-1 isolate had a 99% similarity with Pseudomonas brenneri strain SFML 97-391, and OOH-1 isolate had a 99% similarity with Enterobacter xiangfangensis. Identifcation of antibiotic compounds was done by purifcation and separation of the compounds. Antibiotic activity was also examined based on Lethal Concentration (LC50) on Fusarium oxysporum with a LC50 of 0.01–0.02% against Fusarium oxysporum.
Isolation and Identification of DPPH Radical (2,2-diphenyl-1-pikrylhidrazyl) Scavenging Active Compound in Ethyl acetat fraction of Piper acre Blume Hifdzur Rashif Rijai; Nanang Fakhrudin; Subagus Wahyuono
Majalah Obat Tradisional Vol 24, No 3 (2019)
Publisher : Faculty of Pharmacy, Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/mot.48173

Abstract

Piper acre Blume, known as Black Betel (local name), is a plant that is widely used by the people of East Kalimantan, especially in Samarinda, for the treatment of illness. Leaves (3-4 months old) are collected from Samarinda, extracted, fractionated, and monitored by DPPH antiradical activity. The isolation of the Piper acre Blume is performed on the active fraction, and the structure identification is based on spectroscopic data of the compound.  The leaves were dried, pulverized, and macerated with MeOH. Dried MeOH extract was obtained upon evaporation of the solvent. The extract was then fractionated by vacuum liquid chromatography (vlc), eluted gradually by solvents having different polarities (n-hexane, ethyl acetate and methanol). The fractions obtained were monitored using TLC [n-hexane: ethyl acetate (3: 1 v/v)] that was visualized by UV254 nm, UV366 nm and DPPH. The isolation was performed by preparative TLC [SiO2, n-hexane: ethyl acetate (3: 1)] on ethyl acetate fraction that showed the highest DPPH antiradical value. A single compound was obtained, and it appeared as a round spot and pure according to TLC performances at 3 different solvent systems. The isolated Piper acre Blume compound displayed the IC50 value on the anti-radical DPPH (measured at λ 520 nm) as 10.41µg/mL. The IR spectrum (KBr) showed –OH band (3450 cm-1), aliphatic bands [alkene, 3010 cm-1; alkana 2900 cm-1), an aromatic overtone bands (1900-200 cm-1) and a strong C=O band (1725 cm-1). The NMR (1H- and 13C-) (mono and 2D) indicated the present of a p-di-substituted aromatic signals (δ, 7.54 and 7.52, d, J =6 Hz, 1 H each), 2 methyl (δ, 0.96, d, J = 7.0 Hz, 6 Hs), a triplet signal (δ, 4.22 ppm). Other signals of CH- and CH2 were shown as m signals at δ, 1.64 and 1.34 ppm.  Based on those data, the compound was identified as isoamyl p-OH benzoate that is grouped as parabens used as a preservative in the pharmaceutical preparations. In conclusion, the anti-radical (DPPH) active compound present in the leaves of Piper acre Blume is identified as isoamyl p-OH benzoate, having IC50 value anti-radical DPPH 10,41µg/mL.
Co-Authors Hifdzur Rashif Rija’i Nanang Fakhrudin, Nanang Sebastian Margino Sri Yuwantiningsih