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Indonesian Journal of Biotechnology

ijbiotech
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Published by Universitas Gadjah Mada

ISSN : 08538654 EISSN : 20892241 DOI : -

Core Subject : Science,

Biochemistry, Genetics & Molecular Biology Immunology & microbiology Materials Science & Nanotechnology

The Indonesian Journal of Biotechnology (IJBiotech) is an open access, peer-reviewed, multidisciplinary journal dedicated to the publication of novel research in all aspects of biotechnology, with particular attention paid to the exploration and development of natural products derived from tropical—and especially Indonesian—biodiversity. IJBiotech is published biannually and accepts original research articles featuring well-designed studies with clearly analyzed and logically interpreted results. A strong preference is given to research that has the potential to make significant contributions to both the field of biotechnology and society in general.

Arjuna Subject : -

Articles 530 Documents

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Construction and Immunogenicity Testing of Salmonella, STM1 Vaccine Vector Expressing HIV-1 Antigen Endang Winiati Bachtiar; Peter Smooker; Peter J Coloe
Indonesian Journal of Biotechnology Vol 14, No 2 (2009)
Publisher : Universitas Gadjah Mada

Objective of this study: to determine the ability of Salmonella enterica serovar Typhimurium STM1 as a delivery vehicle for the HIV p24 gene and HIV env gene. The STM1 delivery HIV-p24 vaccination was carried out in the form of a recombinant or a DNA vaccine whereas only a DNA vaccine was used for HIV env. Naked DNA vaccination was also tested and immune responses were evaluated following immunisation in mouse model. Results: vaccination cellular immune responses induced by recombinant p24 STM1 (STM1/pHly-p24) were greater than those elicited by the p24 DNA vaccine in STM1 (STM1/VR-p24), (but statistically not significance) than those induced by oral vaccination. However, IgA responses induced by oral vaccination with either a recombinant or DNA vaccine of p24 in STM1 are higher than those induced by IP vaccination. Conclusions: This result confirms</div><div>other studies that Salmonella was able to deliver HIV antigens to the immune system and induced specific immune responses to the HIV antigen.

Relationship between Nucleus Swelling and Development Competence of Bovine Cloned Embryos Reconstructed by Enucleated Oocytes with Serum-starved or Serum-fed Fetal Somatic Cells Mokhamad Fahrudin; Ni Wayan Kurniani Karja; Tatsuyuki Suzuki
Indonesian Journal of Biotechnology Vol 10, No 2 (2005)
Publisher : Universitas Gadjah Mada

This study was conducted to examine the occurrence of nuclear remodeling (nucleus swelling) and its effects on the subsequent in vitro development of bovine embryos reconstructed by serum-starved and serum-fed somatic cells. Results from this study demonstrated that all of the reconstructed embryos that received serum-starved and serum-fed somatic cells exhibited condensed-nuclei. More than 90% of the transferred nuclei exhibited nuclear envelope breakdown and premature chromatin condensation which clearly distinct from an intact nucleus. There was no significant difference on the degree of nucleus swelling in SS-NT embryos or SF-NT embryos, indicating that either serum-starved or confluent somatic cell lines could be reprogrammed by the recipient cytoplasm environments in similar pattern. Although the fusion rate was not significantly different among the groups, the proportion of SS-NT embryos which developed to the 2- to 4-cell stage (89.7%) and to the 8- to 16-cell stage (74.7%) was significantly higher than that of SF-NT embryos. Whereas, the proportion of reconstructed embryos that developed to the morula and blastocyst stages were not significantly different among the groups. Results of these studies demonstrate that reconstructed embryos, which received either serum-starved or serum-fed confluent somatic cells, showed similar developmental competence to the blastocyst stage.

Combination Methods for Screening Marine Actinomycetes Producing Potential Compounds as Anticancer Yuyun Farida; Jaka Widada; Edy Meiyanto
Indonesian Journal of Biotechnology Vol 12, No 2 (2007)
Publisher : Universitas Gadjah Mada

Marine actinomycetes is a robust source of secondary metabolites including anticancer compounds . The objective of this research was to select marine actinomycetes producing potential compounds as anticancer used combination methods that consist of amplification PKS I (polyketide synthases type I) and NRPS (non ribosomal peptide synthetases) genes, analysis the diversity of secondary metabolites and genetic. Selected isolates were used for cytotoxicity assay. PKS I and NRPS genes were amplified using sets of degenerate primers. K1F and M6R were used for amplify ketosynthase and methyl-malonyl-CoA transferase modules of PKS I gene which targeted sequences 1200-1400 bp. A3F and A7R were used for amplify adenilation domains of NRPS gene which targeted sequences 700-800 bp. The diversity of secondary metabolites was analized by TLC and densitometry of ethyl acetate extracts. Genetic diversity was analized by repetitive DNA fingerprinting using BOXA1R primers. The cytotoxicity of secondary metabolites on T47D and MCF7 breast cell lines cancer was measured by MTT assay method. Fifty two marine actinomycetes isolates were screened using combination methods. Ten isolates were detected encoding both PKS I and NRPS genes, whereas 11 isolates were detected encoding the NRPS gene. The screening by analysis of secondary metabolites and genetic diversity methods were obtained 6 selected isolates for cytotoxicity assay, which consist of 3 isolates encoding both PKS I and NRPS genes and 3 isolates encoding NRPS gene.Isolate 1 had high cytotoxicity with the IC50 on T47D cell was 19 μg/ml and the IC50 on MCF7 cell was 7 g/ml. This findings suggests that combination methods were effective and efficient way to select marine actinomycetes producing potential compounds as anticancer.

Purification and Characterization of Streptomyces sp. IK Chitinase Sebastian Margino; Agustinus Joko Nugroho; Widya Asmara
Indonesian Journal of Biotechnology Vol 15, No 1 (2010)
Publisher : Universitas Gadjah Mada

Streptomyces sp. IK isolated from compost inoculants, could produce extra cellular chitinase in a medium containing 0.2% (w/v) colloidal chitin, fermented for 96 hours at 30oC. The enzyme was purified by a combination of ammonium sulphate precipitation and DEAE-Cellulose anion-exchange chromatography. On SDS-polyacrylamide gel electrophoresis analysis, the purified enzyme showed a mass of 71 kDa. Chitinase was optimally active at pH of 6.7 and at 37oC. Km value and Vmax of the protein for colloidal chitin were 2.92 mg/ml and 4.26 ìg/h, respectively.Key words : chitinase, Streptomyces, purification, characterization

Reactive Oxygen Intermediate (ROI) in Dog Macrophage Infected with Mycobacterium tuberculosis Ida Tjahajati
Indonesian Journal of Biotechnology Vol 11, No 2 (2006)
Publisher : Universitas Gadjah Mada

experiment used 24 healthy dogs, aged between 1 and 2 years, both male and female which were divided into twodifferent groups consisting of 12 dogs each. The first group was the treatment group, that is they were infected with Mtuberculosis and the second one was the control group. The activity of macrophages ROI secretion were measured at1st, 2nd, 12th, and 24th after infection using nitroblue tetrazolium (NBT) reduction assay. Three cats were used to measure themacrophage activity in each period, using triplicate sample for each cat. The results of the experiment showed thatROI secretion increased in infected group compared with the control group, and this activity reached to the plateaulevel at 2 weeks after infection. Although these enhanced activities were gradually diminished thereafter, higherlevels of these activities were consistently observed until the end of experiment compared with control group. Theresults of the experiment indicated that ROI played an important role to against M.tuberculosis infection in dogs.Keyword: macrophage, ROI, M.tuberculosis, dogs

Decolorization of Remazol Briliant Blue R by Laccase from White Rot Fungus Polyporus sp. S133 Tony Hadibarata; Sanro Tachibana
Indonesian Journal of Biotechnology Vol 14, No 2 (2009)
Publisher : Universitas Gadjah Mada

The decolourization of the recalcitrant dye RBBR by the culture filtrate of Polyporus sp. S133 and its isolatedlaccase was investigated. The laccase alone decolorized RBBR. A small molecular weight redox mediator (HBT) wasnecessary to increase the decolorization. The purified laccase totally decolorized the dye of 200 mg l-1 initialconcentration of RBBR when only 1.5 U ml-1 of laccase was used in the reaction mixture. The effects of differentphysicochemical parameters were tested and optimal decolorization rates occurred at pH 5 and at a temperature of 50°C. The effect of surfactants on the decolourization of RBBR was tested with Tween 80, Tween 20, and Brij 35. It wasdemonstrated that Tween 80 was inhibiting substrate for the decolorization while Tween 80 and Brij 35 was noinhibiting effect for the decolorization. Provided that all of the condition is included, it is suggested that laccase maybe suitable for the wastewater treatment of similar anthraquinone dyes.Keywords: Decolorization; Laccase; Remazol Brilliant Blue R (RBBR); Polyporus sp. S133

Mineral Phosphate Solubilizing Bacteria Isolated from Various Plant Rhizosphere under Different Aluminum Content Dolly Iriani Damarjaya; Jaka Widada; Keishi Senoo; Masaya Nishiyama; Shigeto Otsuka
Indonesian Journal of Biotechnology Vol 10, No 2 (2005)
Publisher : Universitas Gadjah Mada

The objectives of this study was to isolate and characterize the mineral phosphate solubilizing bacteria from rhizosphere and evaluate their potential as plant growth promoting bacteria in Al-toxic soils. The halo zone formation method was used to isolate PSB using the media containing insoluble phosphates (Ca-P or Al-P) as a source of phosphate. Eight of acid and Al-tolerant PSB isolates that were able to solubilize Ca-P were obtained from rhizosphere of clover, wheat, corn, and sunflower grown in Al-toxic soil. Identification of the isolates based on the 16S rRNA gene sequence analysis demonstrated that the isolates were strains of Burkholderia (5 strains), Pseudomonas (1 strain), Ralstonia (1 strain), and unidentified bacterium (1 strains). All PSB isolates showed the capability to dissolve Ca-P, and only 1 strain (Ralstonia strain) was able to dissolve Al-P in agar plate medium. The P-solubilization by these isolates was correlated with pH of medium. Inoculation of the bacterial strains on clover on Al-toxic medium showed that all isolates increased the plant dry weight compared with uninoculated treatment. Our results showed that those PSB isolates have potential to be developed as a biofertilizer to increase the efficiency of P-inorganic fertilizer used in Al-toxic soils.

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

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

Effect of Staurosporine on the Intracellular Localization of Hepatitis B Virus Core Protein Aris Haryanto; Nastiti Wijayanti; Michael Kann
Indonesian Journal of Biotechnology Vol 12, No 1 (2007)
Publisher : Universitas Gadjah Mada

protein is also including in the HBV genome targeting into the nucleus through modulating carboxyl residues byphosphorylation. Nuclear localication Signal (NLS) in HBV core protein is inside the virion structure and it must beunmasked in order to function, perhaps by phosphorylation. Phosphorylation of of HBV core protein in turn couldbegin to alter capsid conformation. Staurosporine is a natural product originally isolated from bacteriumStreptomyces staurosporeus. Staurosporine was discovered to have biological activities ranging from anti-fungal toanti-hypertensive. The interest in these activities resulted in a large investigative effort in chemistry and biology andthe discovery of the potential for anti-cancer treatment. The main biological activity of Staurosporine is the inhibitionof protein kinases through the prevention of ATP binding to the kinase. In the present study, we have studied theintracellular localization of EGFP-Core fusion protein with triple HBV core and SV-40 nuclear localization signal atits carboxyl terminal in presence and absence of Staurosporine. We also to study the effect of Staurosporine treatmenton the intracellular localization of EGFP-Core fusion protein in the hepatocyte cells line of HepG2 cell. Resultsshowed that effect of Staurosporine is prevent the nuclear localization of EGFP-Core fusion protein into nucleusthrough an inhibition of the phosphorylation of core protein. Stauroporine also prevents cell division so that passivetrapping of core protein is inhibited.

Cytotoxic activity and apoptosis induction of avocado Persea americana Mill. seed extract on MCF-7 cancer cell line Yuli Widiyastuti; Rarastoeti Pratiwi; Sugeng Riyanto; Subagus Wahyuono
Indonesian Journal of Biotechnology Vol 23, No 2 (2018)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.32141

Avocado Persea Americana Mill. is a commercially important crop and studies have shown that the pulp may have benefits to cardiovascular health, dermatological health and possibly anti-cancer activity. Avocado seeds have several medicinal properties such as anti-hyperglycemic, antimicrobial, antioxidant and anti-inflammation. This study aim to evaluate the effect of avocado seed extract on viability and apoptosis of breast cancer cell line MCF-7. The anticancer effect was evaluated by cytotoxic test using MTT assay and the effect on apoptosis and cell cycle was examined by flow cytometry method. The cytotoxic test showed that chloroform extract had strong cytotoxic activity against MCF-7 cell lines with IC50 value of 94.87 µg/mL. Furthermore, the chloroform extract was partitioned with methanol and yield of soluble methanol fraction (FLM) and non soluble methanol fraction (FTLM). The cytotoxic activity of the methanol soluble fraction (FLM) and non soluble methanol fraction (FTLM) against MCF-7 cell lines was increased with IC50 of 34.52 and 66.03 µg/mL, respectively. Flow cytometry analysis using annexin-V and propidium iodide staining revealed that methanol soluble fraction could induce apoptosis and modulating the cell cycle arrest in MCF-7 cell. This research indicated that avocado seed has a potency to induce apoptosis and as anti-proliferative to MCF-7 cells lines.

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