Abstract. Ardhi A, Sidauruk AN, Suraya N, Pratiwi NW, Pato U, Saryono. 2020. Molecular identification of amylase-producing thermophilic bacteria isolated from Bukit Gadang Hot Spring, West Sumatra, Indonesia. Biodiversitas 21: 994-1000. Amylase is one of the hydrolytic enzymes that is widely used in a wide number of industrial processes such as food, fermentation, textile, paper, detergent, and pharmaceutical industries. Amylase produced by thermophilic bacteria may be thermostable, which is very beneficial in several applications requiring high temperature, for example, the process of gelatinization, liquefaction, and saccharification are performed in high temperature involved in the starch processing. In this study, the amylase-producing ability of thermophilic bacteria isolated from Bukit Gadang hot spring, West Sumatra, Indonesia, was checked and followed by molecular identification. Thirteen isolates that were successfully isolated from the hot springs were microscopically and macroscopically characterized, biochemically tested, and determined their amylase enzyme activity both qualitatively and quantitatively. The isolate that performed the best amylase activity was identified using the molecular technique. The DNA sequencing was carried out in 16S rRNA and continued with BLAST search for species identification. The result of molecular identification showed that the isolate with the best amylase activity was identified as Bacillus licheniformis. The optimum amylase production (231.33 U/ml) and the best enzyme-specific activity (101.79 U/mg) were obtained at the incubation time of 36 hours.
Fusarium oxysporum is one of endophyte microbes isolated from dahlia tuber (Dahlia variabilis) which has the ability to produce secondary metabolites. In this research, optimization of chemichal and physical fermentation conditions were carried out. Corn, potato, and sweet potato with particle size of 80 mesh were used as corbon sources to produce secondary metabolite for 5, 10, 15, 20, and 25 days respectively with ethyl acetate was used to extract secondary metabolites. The assessment of antimicrobial activity was then performed against Candida albican, Escherichia coli and Staphylacoccus aureus. A serial of metabolite concentrations 5.7 mg/ml, 3.8 mg/ml, and 1.9 mg/ml were used. The optimum inhibition against microbial pathogen growth was corn for 15 day fementation (C15). The inhibition zone against Candida albicans, E.coli, and S. aureus were 7.62±0.32, 14.15±0.09, and 15.24±0.24 mm respectively at 5.7 mg/ml metabolite concentration. The result of metabolites screening showed overall extract secondary metabolites comprised terpenoid group. The separation of the active ingredients was performed using both Thin Layer Chromatoghraphy and High Performance Liquid Chromatoghraphy (HPLC) depicted 4 component.
Abstract. Fachrial E, Anggraini S, Harmileni, Nugroho TT, Saryono. 2019. Isolation and molecular identification of carbohydrase and protease producing Bacillus subtilis JCM 1465 isolated from Penen Hot Springs in North Sumatra, Indonesia. Biodiversitas 20: 3493-3498. The application of enzymes industrially is increasing every year and thermophilic microbes are a promising source of these enzymes for industrial use due to their temperature stability. The aim of this study, therefore, was to isolate, characterize and identify the enzyme producer-thermophilic bacteria from the Penen Hot Spring in Deli Serdang, North Sumatra, Indonesia. In the experimental setup, carbohydrase activities including inulinase and amylase were determined by the formation of clear zone around the colonies after soaking with Lugol on the TSA medium supplemented with 1% inulin powder and 1% starch. Similarly, the protease activities were determined by the formation of clear zone around the colonies on Skim Milk Agar Medium. Then, of the 11 isolates, only one known as UTMP 12 showed maximum enzyme activity. The isolate was then characterized based on morphology and biochemistry and found to be Bacillus subtilis strain JCM 1465 (accession number NR_113265, homolog 99.72%). Furthermore, this is the first study on carbohydrase and protease activities of Bacillus subtilis strain JCM 1465 and the result shows that the thermophilic bacteria are needed in the production of carbohydrase and protease.
Fusarium oxysporum LBKURCC41 is an endophytic isolated from Dahlia variabilis tubers from Padang Luar, West Sumatra. The strain was fermented using modified Huangs’ media, ie using corn as a carbon source for 15 days. The purpose of this study is to isolate secondary metabolites from LBKURCC41 strains. The filtrate from the liquid fermentation media was extracted using EtOAc:MeOH at a ratio of 95:5 to 50:50. Purification using fast chromatography was performed with a mixture of BAW (5:1:4). The structure was elucidated by, FTIR and 1H NMR spectroscopy. The isolate was identified as a sesquiterpenoid, with weak antimicrobial activity against E.coli and S. aureus with 2.1 mm clear zone and concentration of 10 ppm.
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