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All Journal Indonesian Journal of Biotechnology Jurnal Jamu Indonesia Indonesian Journal of Jamu
Lucia Dhiantika Witasari, Lucia Dhiantika
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemistry Health Professions Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology Other

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Cloning of Thermostable DNA Polymerase Gene from a Thermophilic Brevibacillus sp. Isolated from Sikidang Crater, Dieng Plateu, Central Java Witasari, Lucia Dhiantika; Prijambada, Irfan Dwidya; Widada, Jaka; Arif Wibawa, Dionysius Andang
Indonesian Journal of Biotechnology Vol 15, No 2 (2010)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (239.792 KB)

Abstract

Thermostable DNA polymerase has an important role for amplifying small amount of DNA through polymerase chain reaction (PCR). Thermophillic bacteria Brevibacillus sp. was isolated from Sikidang Crater, Dieng Plateu, Central Java. Previous study showed that crude protein of the isolate could be used in PCR. Unfortunately, like most native thermostable enzymes, the thermostable DNA polymerase of the isolate is synthesized in a very low level and therefore is cumbersome to purify. The purpose of this research is to clone thermostable DNA polymerase gene of the isolate. The DNA polymerase gene was amplified by means of PCR using spesific primers. The amplified fragment was then isolated, purified, and ligated into the pGEM-T cloning vector. The recombinant plasmid was then transformed to competent E. coli JM109 cells using heat shock method. The cloned thermostable DNA polymerase gene from the thermophilic isolate was then characterized for its nucleotide base sequence. The result showed that the DNA Pol I gene was successfully be amplified from the isolate DNA genom, resulting in ± 2,7 kb DNA fragment in length. Sequence analysis of segment of targeted gene showed high similarity to that of thermostable DNA polymerase genes from other Bacillus.Key words : Thermostable DNA Pol I, Brevibacillus sp., PCR, cloning
Fingerprints, Antioxidant Activity, and Anti-Tyrosinase of Fungus Comb Extract Dewi, Trisni Untari; Diposani, Diva Putri; Rohaeti, Eti; Nandika, Dodi; Karlinasari, Lina; Arinana, Arinana; Santoso, Djoko; Witasari, Lucia Dhiantika; Rachmayanti, Yanti; Batubara, Irmanida
Jurnal Jamu Indonesia Vol. 8 No. 3 (2023): Jurnal Jamu Indonesia
Publisher : Tropical Biopharmaca Research Center, IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/jji.v8i3.346

Abstract

Fungus comb is a structure made by termite colony Macrotermitinae with lots of potential because it contains secondary metabolites such as steroids, terpenoids, and saponins and compounds containing antioxidant and anti-tyrosinase properties. This study aimed to determine fungus comb extracts' fingerprint pattern, antioxidant capacity, and anti-tyrosinase activity. Thin layer chromatography (TLC) was used to determine fingerprint patterns. The resulting TLC pattern was processed with ImageJ and converted into a densitogram. The fungus comb was extracted with n-hexane and ethyl acetate. TLC results showed that fungus comb extracts contain steroids and terpenoids and have antioxidant properties. The antioxidant capacity assay was carried out with 2,2-diphenyl-1-picrylhydrazyl reagent. N-hexane extract had an antioxidant capacity value of 9.07 mg ascorbic acid equivalent (AAE)/g extract, while ethyl acetate extract had 6.04 mg AAE/g extract. Anti-tyrosinase activity assay was carried out with L-tyrosine and L-3,4-dihydroxyphenylalanine (L-DOPA) as substrates. N-hexane extract had a better anti-tyrosinase activity with IC50 of 1804 µg/mL for L-tyrosine and 6784 µg/mL for L-DOPA.
Fingerprints, Antioxidant Activity, and Anti-Tyrosinase of Fungus Comb Extract Dewi, Trisni Untari; Diposani, Diva Putri; Rohaeti, Eti; Nandika, Dodi; Karlinasari, Lina; Arinana, Arinana; Santoso, Djoko; Witasari, Lucia Dhiantika; Rachmayanti, Yanti; Batubara, Irmanida
Jurnal Jamu Indonesia Vol. 8 No. 3 (2023): Jurnal Jamu Indonesia
Publisher : Tropical Biopharmaca Research Center, IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/jji.v8i3.346

Abstract

Fungus comb is a structure made by termite colony Macrotermitinae with lots of potential because it contains secondary metabolites such as steroids, terpenoids, and saponins and compounds containing antioxidant and anti-tyrosinase properties. This study aimed to determine fungus comb extracts' fingerprint pattern, antioxidant capacity, and anti-tyrosinase activity. Thin layer chromatography (TLC) was used to determine fingerprint patterns. The resulting TLC pattern was processed with ImageJ and converted into a densitogram. The fungus comb was extracted with n-hexane and ethyl acetate. TLC results showed that fungus comb extracts contain steroids and terpenoids and have antioxidant properties. The antioxidant capacity assay was carried out with 2,2-diphenyl-1-picrylhydrazyl reagent. N-hexane extract had an antioxidant capacity value of 9.07 mg ascorbic acid equivalent (AAE)/g extract, while ethyl acetate extract had 6.04 mg AAE/g extract. Anti-tyrosinase activity assay was carried out with L-tyrosine and L-3,4-dihydroxyphenylalanine (L-DOPA) as substrates. N-hexane extract had a better anti-tyrosinase activity with IC50 of 1804 µg/mL for L-tyrosine and 6784 µg/mL for L-DOPA.
Co-Authors Arinana Arinana Dionysius Andang Arif Wibawa, Dionysius Andang Diposani, Diva Putri Djoko Santoso Dodi Nandika Eti Rohaeti Irfan Dwidya Prijambada Irmanida Batubara JAKA WIDADA Lina Karlinasari Rachmayanti, Yanti Trisni Untari Dewi