Leaves of jambu air (Syzygium samarangense L.) has been used by local residents as medicine for various diseases caused by free radical agents in human’s body. This study aims to find a new source of antioxidants from medicinal plants and their endophytic fungi Syzygium samarangense leaves was fractionated by maceration method using gradient solvent i.e. n-hexane, ethyl acetate, and methanol. The antioxidant activity of the leaf fractions was determined using 1,1diphenyl-2-picryl hydrazyl (DPPH) method. The chemical compound was isolated from active fraction by chromatographic techniques and their chemical structure was identified using spectroscopy techniques. The endophytic fungi was isolated from S. samarangense leaves and continued to cultivation in potato-dextrose broth (PDB) medium for four weeks. The medium was extracted using ethyl acetate and determined its antioxidant activity. Fungi’s isolate with highest activity was analyzed to identify its molecular. Ethyl acetate fraction of S. samarangense leaves showed highest antioxidant activity. Spectroscopy analysis result concluded the isolated compound is 5,7-dihydroxy-6,8-dimethyl flavanone. Four endophytic fungi had been isolated form S. samarangense leaves. Antioxidant activity test showed that ethyl acetate extract of endophytic fungi BJA-1 has the highest value. Molecular identification of BJA-1 shows high homology with Lasiodiplodia venezuelensis strain CBS 129753.
Microbial communities in an acidic hot spring, namely Kawah Hujan B, at Kamojang geothermal field, West Java-Indonesia was examined using culture dependent and culture independent strategies. Chemical analysis of the hot spring water showed a characteristic of acidic-sulfate geothermal activity that contained high sulfate concentrations and low pH values (pH 1.8 to 1.9). Microbial community present in the spring was characterized by 16S rRNA gene combined with denaturing gradient gel electrophoresis (DGGE) analysis. The majority of the sequences recovered from culture-independent method were closely related to Crenarchaeota and Proteobacteria phyla. However, detail comparison among the member of Crenarchaeota showing some sequences variation compared to that the published data especially on the hypervariable and variable regions. In addition, the sequences did not belong to certain genus. Meanwhile, the 16S Rdna sequences from culture-dependent samples revealed mostly close to Firmicute and gamma Proteobacteria.
Abstract. Fadhillah, Elfita, Muharni, Yohandini H, Widjajanti. 2019. Chemical compound isolated from antioxidant active extract of endophytic fungus Cladosporium tenuissimum in Swietenia mahagoni leaf stalks. Biodiversitas 20: 2645-2650. Swietenia mahagoni L. Jacq is a medicinal plant that widely used as an antimicrobial, anti-inflammatory, antimutagenic, anticancer, antitumor, and antidiabetic. Several bioactive compounds have been reported to be associated with this plant that supports its treatment activities. Flavonoid content of the S. mahagoni has antioxidant activity. Therefore the plant is widely used to treat degenerative diseases caused by free radical activity in the body. Scientists have proven that the medicinal plants have a symbiotic relationship with the number of microorganisms that can produce bioactive compounds. One of the symbiotic microorganisms is endophytic fungi believed to be associated with every plant on earth. In this paper, we report a secondary metabolite compound from endophytic fungi isolated from S. mahagoni. The endophytic fungi (DMG1-DMG4) were isolated from leaf stalks of S. mahagoni. Antioxidant activity of endophytic fungi extracts was tested by 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. The chemical compound from the active extract was isolated by the chromatographic method, and the structure was elucidated by spectroscopic methods including UV, IR, NMR 1D (1H-NMR and 13C-NMR), NMR 2D (HMQC and HMBC). Antioxidant activity test showed the endophytic fungus (DMG3) has the highest activity. The chemical compound from DMG3 was identified as 5-hydroxy-2-oxo-2H-piran-4-yl) methyl acetate). The phylogenetic method used to molecular analysis of DMG3 confirmed the fungus as Cladosporium tenuissimum.
A novel thioesterse gene was successfully cloned and sequenced directly from natural sample of Domas Hot Spring, West Java, Indonesia. Homological analysis of the sequence showed that the gene appeared high homology to thioesterase genes with the highest to a putative thioesterase gene from uncultured Acidilobus sp. JCHS at 66% identity. However, phylogenetic analysis showed that the protein was separated from the branch with other known thioesterases. The size of the gene is around 500 base pairs, lied into 2 kb DNA fragment from a random PCR amplicon. The gene was overexpressed in Escherichia coli , a dominant band appeared at 17 kDa in SDS-PAGE with expression level at around 32% of total proteins. The activity of the purified protein using acetyl-CoA as substrate showed that the protein exhibited thioesterase activity. Furthermore, the enzyme also showed esterase activity on p-nitrophenyl ester as substrate. Detail characterization of esterolytic activity showed that the enzyme preferred p-nitrophenyl decanoate as substrate. The optimum activity of the enzyme was at 80 °C and pH 8. Activity of the enzyme was maintained after incubation at 80 °C up to 24 h. In addition, the enzyme was favorable on polar organic solvents. All the data obtained suggested that the enzyme is a novel alkaline thermostable thioesterase.
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