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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.

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Articles 523 Documents

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Diversity of Actinomycetes at Several Forest Types in Wanagama I Yogyakarta and Their Potency as a Producer of Antifungal Compound Reni Nurjasmi; Jaka Widada; N. Ngadiman
Indonesian Journal of Biotechnology Vol 14, No 2 (2009)
Publisher : Universitas Gadjah Mada

Actinomycetes are bacterial groups that produce many secondary metabolites, which different biological activities, such as antifungi, antibacteria, antivirus, antitumor, etc. Actinomycetes are widely distributed in soil and their diversity is influenced by type of forest. The aim of this study is to investigate diversity of actinomycetes in several forest types of Wanagama I forest in Yogyakarta and their potency as a producer of antifungal compound. Soil samples under the forest of Tectona grandis, Swietenia macrophylla King, Bamboosa vulgaris, Melaleuca leucadendron, and Gliricidia maculata were used as sources of soil bacteria. Bacteria and actinomycetes communities were analyzed through culture-independent approach by RISA and nested-PCR RISA using actinomycetes spesific primer (F243), respectively. Through culture-dependent approach, isolated actinomycetes diversity were analyzed by identification of morphology (colony and cell), genetic (BOX element by rep-PCR), and secondary metabolites (thin layer chromatography). In addition, isolates were assayed for their antifungal activity against Saccharomyces cerevisae, Candida albicans, Fusarium oxysporum and Aspergillus flavus. The presence of Polyketide Synthase-I (PKS-I) and NonRibosomal Peptide Synthetase (NRPS) genes were amplified by PCR to study their correlation with antifungal activity of the actinomycete isolates. The results showed that types of forest influence diversity of rhizobacteria especially actinomycetes. According to culture-independent approach, relatively, com-</div><div>munity of rhizobacteria from the highest were soil under the forest of B. vulgaris, G. maculata, T. grandis, S.macrophylla King, and M. leucadendron, respectively. Meanwhile, community of actinomycetes from the highest were soil under the forest of G. maculata, B. vulgaris, M. leucadendron, S. macrophylla King, and T. grandis, respec- tively. Fourty-three morphologically different isolates were found by using culture-dependent approach consisting of 17 isolates were found in soil under the forest of M. leucadedron, each of 9 isolates in G. maculata and T. grandis, 6 isolates in S. macrophylla King. and 2 isolates in B. vulgaris. More diversity of secondary metabolites were observed in soil actinomycetes under the forest of M. leucadendron. Of the 43 isolates, 100% were active against S.cerevisae, 37.20% against C. albicans, 95.30% against F. oxysporum, and 83.70% against A. flavus. Antifungal activity of actinomycete isolates did not always have correlation with the presence of PKS-I and NRPS.

Expression of haloacid dehalogenase gene and its molecular protein characterization from Klebsiella pneumoniae ITB1 Ridani Rino Anggoro; Enny Ratnaningsih
Indonesian Journal of Biotechnology Vol 22, No 1 (2017)
Publisher : Universitas Gadjah Mada

Organohalogen compounds are widely used industrially and agriculturally, as well as in households as flame retardants and refrigerants. However, these compounds can become significant pollutants through their accidental or deliberate release into the environment in large quantities. Dehalogenase is an enzyme with the potential to be used in the removal of organohalogen contaminants. A previous study successfully subcloned a 690 bp of haloacid dehalogenase gene (hakp1) from Klebsiella pneumoniae ITB1 into a pET-30a(+) expression system to achieve high enzyme productivity. IPTG was used as an inducer to express a pET-hakp1 recombinant clone in Escherichia coli BL21 (DE3). The molecular mass of the haloacid dehalogenase Hakp1 protein was 30 kDa as determined by SDS-PAGE. Zymogram analysis showed that this recombinant protein has dehalogenase activity as shown by the formation of AgCl white precipitate. A quantitative assay of haloacid dehalogenase Hakp1 gave a specific activity of 84.29 U/mg with the optimum temperature of 40°C at pH 9. Predicted three-dimensional structure of Hakp1 showed α/β motif folding which comprised of cap and core domain. The predicted active sites of Hakp1 were Asp8, Glu10, Leu22, Phe23, Trp90, Ser125, Ser126, Lys159, and Asp184 with Asp8, Glu10, Ser126, and Lys159 act as binding residue. This recombinant haloacid dehalogenase clone provides an alternative agent for effective bioremediation of organohalogen pollutants.

Phylogenetic relationship of Gram Negative Bacteria of Enterobacteriaceae Family in the Positive Widal Blood Cultures based on 16S rRNA Gene Sequences Sri Darmawati; Langkah Sembiring; Widya Asmara; Wayan T. Artama; Masashi Kawaichi
Indonesian Journal of Biotechnology Vol 19, No 1 (2014)
Publisher : Universitas Gadjah Mada

The purpose of this study was to analyze the phylogenetic relationship of Gram negative bacteria (3strains of Salmonella typhi, 1 strain of Escherichia coli, 1 strain of Serratia marcescens, and 3 strains of Enterobactercloacae) of Enterobacteriaceae family in positive Widal blood cultures based on 16S rRNA gene sequences. Theresults respectively showed that each two 16S rRNA gene clones of Serratia marcescens KD 08.4 had a closerelationship with 16S rRNA gene of Serrratia marcescens ATCC 13880 (similarity: 99.53-99.8%), Eschericia coliBA 30.1 with Eschericia coli ATCC 11775T (similarity: 99.38-99.67%), Salmonella typhi BA 07.4, Salmonella typhiKD 30.4, and Salmonella typhi SA 02.2 with Salmonella typhi ATCC 19430T (similarity: 99.4-100%) as well as theisolates of Enterobacter cloacae SA 02.1, Enterobacter cloacae BA 45.4.1, one 16S rRNA gene clone of Enterobactercloacae TG 03.5 with Enterobacter cloacae ATCC 23373 (similarity: 99.0-99.87%).

Regression Analysis for the Identification of RAPD Markers Linked To Drought Tolerance in Sorghum Paramita Cahyaningrum; T. Taryono; Anto Rimbawanto
Indonesian Journal of Biotechnology Vol 17, No 1 (2012)
Publisher : Universitas Gadjah Mada

Sorghum (Sorghum bicolor) can actually withstand in dry or drought condition better than other crops,therefore it can be grown at different agroclimatic conditions and its product can be used for different purposessuch as food, feed and industrial raw material. However at severe condition, the productivity will also dropdrastically. The aim of this research was to identify RAPD marker linked to the drought tolerance. In thisresearch, varieties of sorghum used as research materials were Durra, Zhengzu, the mutants of Durra andZhengzu (from 300 Gy gamma radiation) B-100 and Zh-30, and the F2 seeds from Zh-30 x B-100 and B-100 xZh-30. Drought screening was carried out using 0.3 % KI during sorghum vegetative stage. DNA extractionwas done using a modified CTAB method. PCR was carried out for RAPD analysis. PCR amplification productswere scored and analyzed using SAS program. The result showed that potassium iodide can be used fordrought screening during the vegetative stage and regression analysis using the logistic method can be usedto identify RAPD markers that is linked to drought tolerance in sorghum. The logistic analysis showed thatband A8-480 was linked to drought tolerance in sorghum.

Citrus reticulata's Peels Modulate Blood Cholesterol Profile and IncreaseBone Density of Ovariectomized Rats Rosa Adelina; Maria Dwi Supriyati; Dwi Ana Nawangsari; Riris I Jenie; Edy Meiyanto
Indonesian Journal of Biotechnology Vol 13, No 2 (2008)
Publisher : Universitas Gadjah Mada

Hormon Replacement Therapy is a common therapy for estrogen deficiency but in other side it will increase the risk of cardiovascular disease. Another alternative therapy which relatively more safe is using phytoestrogen. The Citrus reticulata’s peel contain flavanone and polimethoxyflavone which are suspected to give estrogenic effect, therefore it is potential to be used as phytoestrogen.The purpose of this study was to examine the estrogenic effect of Citrus reticulata’s peel extract in modulation of bone density and blood cholesterol profile of ovariectomized rats (OVX), an animal model of postmenopausal osteoporosis. Thirty six 7-weeks-old female Sprague Dawley rats were assigned to six groups: a SO group, an OVX group, an OVX+CMCNa group, an OVX+extract dose 500 mg/kgBW group, an OVX+extract dose 1000 mg/kgBW group, and an OVX+estradiol group. After 7 weeks, the rats were killed then blood and femoral were collected immediately. The rontgenogram indicated that extract and estradiol administration increase the bone density. And the data analysis with Oneway ANOVA test ,followed by Shceffé test (P 0.05) showed that extract can improve blood cholesterol profile in dose depend manner. These results suggest a possible role of Citrus reticulata’s peel extract as women’s health agent because of its beneficial effects on bone and lipids.

Analysis of whole cell protein profiles by SDS-PAGE to identify indigenous cellulose-producer acetic acid bacteria Sarkono Sarkono; Soekarti Moeljopawiro; Bambang Setiaji; Langkah Sembiring
Indonesian Journal of Biotechnology Vol 21, No 2 (2016)
Publisher : Universitas Gadjah Mada

This study was carried out to analyze the suitability of the identification of four indigenous cellulose-producing acetic acid bacterial isolates (ANG29, KRE65, ANG32 and SAL53) based on the analysis of whole cellular protein profiles against identification based on phenotypic traits. Whole cellular protein profiles were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) method. The whole cellular protein profiles obtained from sample isolates, were compared with reference isolates for species identification. The results showed that based on visual observations can be determined as much as 12 bands of protein with a molecular weight of 19,099 KDa up to 132.182 KDa. Based on the analysis of protein bands were detected visually, fourth indigenous cellulose- producing acetic acid bacterial isolates in the study had a higher similarity profile to the reference strain Gluconacetobacter xylinus BTCC 769 compared with other reference strains namely G. hansenii NBRC 14820T. This condition is consistent with the results of the identification of fourth cellulose producing acetic acid bacterial isolates based on phenotypic traits. Thus, the whole cellular protein profiles by SDS-PAGE technique can be used as a one of method to identification of cellulose producing acetic acid bacterial isolates.

Human Origin Lactobacillus casei Isolated from Indonesian Infants Demonstrating Potential Characteristics as Probiotics in vitro W. Widodo; Tiyas Tono Taufiq; Ety Aryati; Asih Kurniawati; Widya Asmara
Indonesian Journal of Biotechnology Vol 17, No 1 (2012)
Publisher : Universitas Gadjah Mada

The aim of this experiment was to isolate and identify Lactic Acid Bacteria (LAB) from infant faecesand subsequent evaluation of its potential probiotics. LAB was isolated from faeces of infants who consumedbreast milk as the only source of diet on L-cysteine-supplemented MRS Agar, and incubated on 37oC for 48hours. Colonies grew on this media were then identifi ed based on morphological, physiological and molecularapproaches. Morphological and physiological identifi cations based on Gram staining, shape, motility, sporeformation, catalase, CO2 and NH3 production, and the ability to grow on temperature at 10oC and 45oC.Molecular identifi cation based on the amplifi cation of 16S rRNA gene. The potential application of selectedisolates for probiotics was evaluated based on the ability to grow on media with low pH and the additionof 0.5% bile salts, the ability to inhibit the growth of pathogenic Bacillus cereus and Eschericia coli, and in vitroadherence ability. On the basis of morphological, physiological and molecular analysis of 16S rRNA gene, itwas concluded that the selected isolate 1AF was a strain of Lactobacillus casei. Evaluation of probiotic in vitro showed that 60.4% of cells were resistant to pH 2.0 for 90 minutes. Survival of isolate 1AF after growing at0.5% bile salts was 70.8%. The selected isolate 1AF showed the ability to inhibit the growth of Eschericia coli and Bacillus cereus with inhibitory zone of 12.00±1,00 and 15.33±1.53 mm, respectively. In vitro study on theadherence value of isolate to solid plate was found at 46.5%. It is concluded that Lactobacillus casei isolate 1AFis a potential candidate as probiotics and subject to further in vivo evaluation.

Determination of secondary and tertiary structures of cervical cancer lncRNA diagnostic and siRNA therapeutic biomarkers Arli Aditya Parikesit; Didik Huswo Utomo; Nihayatul Karimah
Indonesian Journal of Biotechnology Vol 23, No 1 (2018)
Publisher : Universitas Gadjah Mada

Cervical cancer is one of the primary causes of mortality in women due to human papilloma virus (HPV) infection. The fingerprint of an HPV infection could be detected using a long non-coding RNA (lncRNA) biomarker, enabling it to be utilized in molecular diagnostics. The primary structure or sequences of RNA should be annotated within conventional bioinformatics tools. Therefore, this study aimed to determine the fine-grained 2D and 3D structures of lncRNA PVT1 and its respective siRNA inhibitors. lncRNA PVT1 sequences from Homo sapiens, Mus musculus, and Rattus norvegicus were retrieved from Genbank (NCBI). Prediction of the 2D structure and analysis of the interactions of the lncRNA and siRNA were performed using the Vienna RNA package. The 3D structure of the RNA was computed using the SimRNA and ModeRNA software programs. The results showed that lncRNA PVT1 from H. sapiens and M. musculus had a conserved region. However, the lncRNA from both H. sapiens and M. musculus showed a low conserved region, and the 2D structure could not be determined; thus, the annotation and 2D model focused only on H. sapiens. Both of their lncRNA PVT1 also had a short half-life in the cell. Based on the 3D modeling pipeline, the 3D model of lncRNA PVT1 showed the stability and possible function as molecules, while the PVT1 siRNA-lncRNA interaction analysis revealed that the molecules could bind well. Based on these findings, the structures of both lncRNA PVT1 and its siRNA have the potential to be utilized as biomarkers.

The Phylogenetic Relationship Among Varieties of Lansium domesticum Correa Based on ITS rDNA Sequences Laila Hanum; Rina Sri Kasiamdari; S. Santosa; R. Rugayah
Indonesian Journal of Biotechnology Vol 18, No 2 (2013)
Publisher : Universitas Gadjah Mada

Lansium domesticum Corr. with vernacular name in Indonesian duku has been reported containingtherapeutic bioactive compounds, and some of these compounds shown to be potent antitumor, anticancer,antimalaria, antimelanogenesis, antibacteria, and antimutagenic activities. This plant is commonly known asduku, kokosan and langsat by the local community in Indonesia. The morphological appearance of all varieties isnearly the same, and identifi cation of the varieties is very diffi cult for growers. Variation of DNA sequences ofthe ITS (Internal transcribed spacer) region can be used as a molecular character to determine the phylogeneticrelationship of different varieties of L. domesticum. The aims of this study were to determine taxonomy status ofduku, kokosan, and langsat, also phylogenetic relationship among varieties of L. domesticum based on ITS rDNAsequencing. DNA was isolated from leaves of plant and then amplifi ed using F1 and R1 primers. Nucleotidesequences were identifi ed using Sequence Scanner Software Programm version 1.0, nucleotide sequences from18S, ITS1, 5.8S, ITS2 and 26S region, that has been mergered using EditSeq and SegMan in software Suite forSequence Analysis DNASTAR Lasergene DM version 3.0.25. The results of study showed that DNA fragmentsranging in size from 782-810 bp. Different pattern of DNA fragments indicated polymorphism among duku,kokosan, and langsat. Based on the results of the ITS rDNA sequencing and phylogenetic tree analysis. Itwas determined that Lansium and Aglaia are a separated genus with the similarity index value of 0.98. Duku,kokosan and langsat were divided into two cluster, namely cluster kokosan-langsat and cluster duku with thesimilarity index value of 0.996. Keywords : Phylogenetic relationship, ITS region, L. domesticum, duku, kokosan, langsat

Diversity of Nonribosomal Peptide Synthetase Genes in the AnticancerProducing Actinomycetes Isolated from Marine Sediment in Indonesia Camelia Herdini; Shinta Hartanto; Sofia Mubarika; Bambang Hariwiyanto; Nastiti Wijayanti; Akira Hosoyama; Atsushi Yamazoe; Hideaki Nojiri; Jaka Widada
Indonesian Journal of Biotechnology Vol 20, No 1 (2015)
Publisher : Universitas Gadjah Mada

Marine actinomycetes is a group of bacteria that is highly potential in producing novel bioactivecompound. It has unique characteristics and is different from other terrestrial ones. Extreme environmentalcondition is suspected to lead marine actinomycetes produce different types of bioactive compoundfound previously. The aim of this study was to explore the presence and diversity of NRPS genes in 14anticancer-producing actinomycetes isolated from marine sediment in Indonesia. PCR amplificationand restriction fragment analysis of NRPS genes with HaeIII from 14 marine actinomycetes were doneto assess the diversity of NRPS genes. Genome mining of one species of marine actinomycetes (strainGMY01) also was employed towards this goal. The result showed that NRPS gene sequence diversity in 14marine actinomycetes could be divided into 4 groups based on NRPS gene restriction patterns. Analysisof 16S rRNA gene sequences of representatives from each group showed that all isolates belong to genusof Streptomyces. Genome mining result showed that strain GMY01 harboring 10 different NRPS geneclusters that encode secondary metabolites, as pure NRPS or hybrid between NRPS and other compounds.These results indicated that marine actinomycetes having a high potential to be developed as source ofanticancer drugs development.

Page 24 of 53 | Total Record : 523

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