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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 20 Documents

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Search results for , issue "Vol 16, No 2 (2011)" : 20 Documents clear

Effect of Oxidative Stress on AhpC Activity and Virulence in katG Ser315 Thr Mycobacterium tuberculosis Mutant Ning Rintiswati; Tri Wibawa; Widya Asmara; Hardyanto Soebono
Indonesian Journal of Biotechnology Vol 16, No 2 (2011)
Publisher : Universitas Gadjah Mada

Mycobacterium tuberculosis strains resistance to INH is mainly caused by the alteration in several genesencoding the molecular targets. Mutation of katG at codon 315 especially Ser315Thr are responsible forINH resistance in a large proportion of TB cases. The aim of this study is to evaluate the influence of stressoxidative on AhpC activity of katG Ser315Thr of M.tuberculosis, and to find out the relation of AhpC and thevirulence of this mutant. The study design was laboratoric experimental, subjects of study were M.tuberculosisINH resistance strains, and the treatment were serial dose of H2O2. Eighty five M.tuberculosis INH resistantclinical strain were screened for mutation of katGSer315Thr by PCR/RFLP and characterized on the basis ofphenotypic properties (catalase activity and AhpC activity). AhpC activity of katG Ser315Thr M.tuberculosisstrains in response to oxidative stress condition was evaluated by culturing the strains on liquid culturemedium containing 1mM H2O2.To ascertain role of AhpC in the virulence of katGSer315Thr mutant strains, themutants were infected into human macrophages culture, and several indicator of virulence were observed (i.e:replication competence, and apoptosis induction on human macrophages). The results showed that katG Ser315Thr were identified in 23 (27,05%) of 85 INH resistance strains, all mutant strains had decrease of catalaseactivity. AhpC activity of katG Ser315Thr of M.tuberculosis increased significantly with increase of hydrogenperoxide dose. In addition , it has been shown that increased AhpC activity related to replication ability ofmutant, and reduction of apoptosis macrophages induction significantly. We conclude that the productionof AhpC of katG Ser315Thr M.tuberculosis induced by oxidative stress. There was a role of AhpC in virulenceof the M.tuberculosis katG Ser315Thr strains by replication capability and macrophages apoptosis.

Genetic Variation of Apolipoprotein E (ApoE) in Surabaya, Palu and Alor Populations of Indonesia Pramudji Hastuti; Abdul Salam Mudzakir Sofro; Ahmad Husain Asdie; Ahmad Hamim Sadewa
Indonesian Journal of Biotechnology Vol 16, No 2 (2011)
Publisher : Universitas Gadjah Mada

Apolipoprotein E (ApoE) has been considered to play an important role in cardiovascular disorders.Several studies reported that genetic variation in ApoE locus influence plasma lipoprotein level. The objectivesof this study was to compare the frequency of ApoE genotypes and alleles in some populations of Indonesia.One hundred and ninety five voluntarily unrelated apparently healthy individuals were recruited fromSurabaya, Palu and Alor representing the western, middle and eastern populations of Indonesia, respectively.Blood samples were collected from each subject for DNA extraction. The common allelic variants of ApoE werescreened using polymerase chain reaction (PCR) and restriction fragment length polymorphism. Three allelesi.e. ε2, ε3 and ε4 were identified and five genotypes i.e. ApoE ε2/ε2, ApoE ε2/ε3, ApoE ε3/ε3, ApoE ε2/ε4, ApoE ε3/ε4 were found in three populations studied, while ApoE ε4/ε4 was absent in Surabaya, representing the westernpopulations of Indonesia. The frequency of ε2, ε3 and ε4 alleles in the western population were 0.208, 0.701and 0.092 respectively; in the middle population were 0.242, 0.618 and 0.140 respectively and in the easternpopulation of Indonesia were 0.267, 0.466 and 0.267 respectively. The highest frequency of ε2 and ε4 allelewas found in the eastern population of Indonesia. The distribution of ε2 allele were not significantly differentamong all Indonesian populations, but significantly different were found in ε3 and ε4 allele in the easternpopulation compared to those in the western and middle populations of Indonesian. It can be concluded thatthe frequency of three ApoE alleles in the western and middle populations of Indonesia was not significantlydifferent however, significantly different was observed in the frequency of ApoE ε3 and ε4 alleles from theeastern compared to those in the western and middle populations of Indonesia.

Ability of Curcuminoid from Curcuma domestica Val. in Reducing the Secretion of Reactive Oxygen Intermediates by Synovial Fluid Monocytes in Patients with Osteoarthritis Nyoman Kertia; Ahmad Husain Asdie; Wasilah Rochmah; M. Marsetyawan
Indonesian Journal of Biotechnology Vol 16, No 2 (2011)
Publisher : Universitas Gadjah Mada

Increasing the secretion of reactive oxygen intermediates (ROI) by monocytes in the synovial fluid is anindicator to determine the severity of joint inflammation. Previous studies have shown that curcumin inhibitthe osteoarthritis progression with its ability to inhibite the activity of the nitric oxide synthase (NOS) enzymefrom macrophages. In this prospective randomized open end blinded evaluations = PROBE study, 80 patientswith knee osteoarthritis were eligable. The subject were devided in to two group: group who received 3 x 30mg of curcuminoid from Curcuma domestica Val. extract (curcuminoid group) and group who received 3 x 25mg of diclofenac sodium (diclofenac group) as comparison. The treatment was for 4 weeks time. The secretionof ROI by sinovial fluid monocytes was calculated by scoring the amount of formazan formation after neutralred staining in nitrobleu tetrazolium reduction assay. The result of this study showed that the secretion of ROIby synovial fluid monocytes was significantly decreased in both groups (p <0.001) respectively. There wasno significant difference in decreasing of ROI secretion of synovial fluid monocytes between both treatmentgroups (p = 0.92).

T-786c Polymorphism in nitric oxide synthase 3 gene and Nitrit Oxide Level of Diabetic Retinopathy in Javanese Population Putri Widelia Welkriana; S. Sunarti; Pramudji Hastuti
Indonesian Journal of Biotechnology Vol 16, No 2 (2011)
Publisher : Universitas Gadjah Mada

Complication of retinopathy in type 2 DM is caused of lower level of NO. Nitric oxide level is synthesizedfrom L-arginin in reaction that catalyze Nitric oxide synthase (NOS) 3. The T-786C mutation in NOS 3 genedecreases the expression of nitric oxide synthase (NOS) 3 so decreases NO synthesis. To investigate theassociation between T-786C polymorphism in NOS 3 gene with NO level of diabetic retinopathy patients. Thisstudy was a case control study, consist of 40 patient of type 2 diabetic with DR (case group) and 40 patient oftype 2 diabetic without DR (control group) of Javanese ethnic. The genotyping of T-786C polymorphism wasperformed by PCR-RLFP. Level of NO was measured by spectrophotometry. Chi square test and odd ratiowere used to analyze the association of the T-786C polymorphism in NOS 3 gene with DR. Differences ofNO level between TT and TC genotypes were analyzed using independent t test. The distribution of T-786Cpolymorphism in NOS 3 gene of DR subjects showed that frequency of TT genotype was 22.5% and TC genotypewas 77.5%. Non DR subjects showed the frequency of TT genotype was 50% and TC genotype was 50%, (p=0.011). Frequency of T allele in DR group was 61.25% and C allele was 38.75%, and frequency of T allele in nonDR group was 75% and C allele was 25%, (p= 0.62). Odd ratio of TC genotype was 3.444(CI; 95% : 0.964-3.735)and C allele was 1.898 (CI; 95% : 1.310-9.058). The NO level of TC genotype was 1.43+0.126 and TT genotypewas 11.27+5.87 (p=0.000). Level of NO between RD and non RD showed not different significantly (p=0.160)for retinopathy. The T-786C polymorphism of NOS 3 gene is risk factor for retinopathy in type 2 DiabetesMellitus. Individual with TC genotype of NOS 3 gene has lower level of NO than TT genotype.

Determination of Haemaglutinin and Gene Encoding Fibronectin Binding Proteins Staphylococcus aureus Isolated from Dairy Milk Cows Feny Prabawati Pratomo; Siti Isrina Oktavia Salasia; Syarifudin Tato
Indonesian Journal of Biotechnology Vol 16, No 2 (2011)
Publisher : Universitas Gadjah Mada

Staphylococcus aureus is a major pathogen causing clinical and subclinical mastitis in dairy milk cows. Themastitis has immense economical impacts, where it reduces of the quantity and quality of milk production.The aims of the research were to analyse haemaglutinin and gene encoding fbronectin binding proteins.Nineteen Staphylococcus aureus isolates used in the present study were isolated from dairy milk cows fromYogyakarta, Solo, Boyolali and Sumedang. The haemagluitinin of S. aureus were determined based onhaemaglutination reaction to erythrocytes of rabbit. Detection of gene encoding fbronectin binding proteinscould be performed with specifc primers using polymerase chain reaction (PCR). The results of studiesshowed that most of S. aureus (78,95%) expressed haemaglutinin based on their ability to aglutinate rabbiterythrocytes. Analysis of gene encoding fbronectin binding proteins of S. aureus revealed gene fnbA withsize of approximately 1300 bp for 57,89% isolates, gene fnbB with size of approximately 900 bp for 31,58%isolates and both of gene fnbA and fnbB could be detected for 31,58% isolates. The characters of S. aureusbased on haemaglutinin, gene fnbA and fnbB of the present study could be used as an information to controlof S. aureus infection in dairy herds.

CYP3A4*1G gene Polymorphism on Javanese People Em Sutrisna; Iwan Dwiprahasto; Erna Kristin
Indonesian Journal of Biotechnology Vol 16, No 2 (2011)
Publisher : Universitas Gadjah Mada

Most of drugs are metabolized by cytochrome P 450 (CYP) enzyme. Cytochrome P450 3A4 is thecytochrome that is involved in metabolizing more than 60% of all medicine used in human. The variationof this CYP3A4 gene will affect the catalytic activity of this enzyme. Recently, CYP3A4*1G in intron 10 wasfound in Chinese and Japanese population. There is a substitution of G to A at position 82266 in intron 10. Thepurpose of this research was to investigate the frequency of allele and genotype CYP3A4*1G. Samples weretaken from bloods of the subjects of the research. The examination of CYP3A4*1G was conducted by RTLP-PCRmethod.As the results of this research, the frequency of CYP3A4*1G in Javanese people is CYP3A4*1/*1 0.25,CYP3A4*1/*1G 0.55 and CYP3A4*1G/*1G 0.20. Frequency of allele G: 0.53, allele A: 0.47. The Fisher’s exact- testshows that the allele and genotype frequencyis p. 1.000. The allele and genotype frequency of Javanese peopleisstill in Hardy-Weinberg equilibrium.

Early Detection and Serotyping of Dengue Viruses Clinical Isolates Using Reverse Transcription Polymerase Chain Reaction (RT-PCR) 2 Primers Abdul Rahman Siregar; Tri Wibawa; Nastiti Wijayanti
Indonesian Journal of Biotechnology Vol 16, No 2 (2011)
Publisher : Universitas Gadjah Mada

Recently several methods for confirming Dengue Virus have been developed involve virus isolation, detection of virus antigen, and nucleic acid using PCR. It has been reported that rapid detection method for confirming DHF by Multiplex RT-PCR had been successfully developed. It was more effective than the other methods with a high sensitivity and specivicity were 100% at the early phase (day 1-3). This study was designed to develop rapid detection and serotyping methods for Dengue Virus using RT-PCR 2 primers (Dcon and preM) with annealing temperature was 57oC. The whole blood samples were collected from suspected dengue fever patients that had been confirmed with NS1 kit from R.S. Persahabatan DKI Jakarta and R.S. Prof. Dr. Sardjito DI Yogyakarta during Februari-August 2009. The PCR products showed that in 12 samples, 100 % were postitive with different pattern among the serotypes especially for DEN1 and DEN2, but not for DEN3 and Den4. This method was also able to confirm the double infection DEN2-DEN3, but not for the other ones because of the unspecific pattern. From the results, it indicated that the 2 primers can be a promising early detection and serotyping method of Dengue Virus which infected the DHF patients. Key words: Dengue Virus, DHF, early detection, serotyping, RT-PCR 2 primers.

Molecular Identifcation of Lactic Acid Bacteria Producing Antimicrobial Agents from Bakasang, An Indonesian Traditional Fermented Fish Product Helen Joan Lawalata; Langkah Sembiring; Endang Sutriswati Rahayu
Indonesian Journal of Biotechnology Vol 16, No 2 (2011)
Publisher : Universitas Gadjah Mada

Twenty seven strains of lactic acid bacteria (LAB) were isolated from bakasang, Indonesian traditionalfermented fsh product. In general, LAB have inhibitory activity againts pathogenic bacteria and spoilagebacteria. Screening for antimicrobia activity of isolates were performed with well-diffusion method. Oneisolate that was designed as Pediococcus BksC24 was the strongest against bacteria pathogenic and spoilagebacteria. This strain was further identifed by 16S rRNA gen sequence comparison. Isolates LAB producingantimicrobial agents from bakasang were identifed as Pediococcus acidilactici.

Biodegradation of Used Engine Oil by Acinetobacter junii TBC 1.2 Witono Basuki; Khairul Syahputra; Ayu Tri Suryani; Ilham Pradipta
Indonesian Journal of Biotechnology Vol 16, No 2 (2011)
Publisher : Universitas Gadjah Mada

The isolates have capability to degrade used engine oil was obtained from soil in the beach contaminatedwith used engine oil. One of the selected isolates TBC 1.2 was identified by its 16s rDNA as Acinetobacterjunii. The microorganism can use hydrocarbons in used engine oil as the sole carbon source and energy, alsoit significantly degraded almost all hydrocarbon compounds in used engine oil. With its ability Acinetobacterjunii TBC 1.2 has a potency to be utilized for bioremediation of soil polluted with used engine oil.

Development of Random Amplified Polymorphism DNA Markers Linked to Powdery Mildew Resistance Gene in Melon Budi Setiadi Daryono; Ganies Riza Aristya; Rina Sri Kasiamdari
Indonesian Journal of Biotechnology Vol 16, No 2 (2011)
Publisher : Universitas Gadjah Mada

A random amplified polymorphic DNA (RAPD) marker linked to powdery mildew resistance gene (Pm-I) in melon PI 371795 was reported. However, the RAPD marker has problem in scoring. To detect powdery mildew resistance gene (Pm-I) in melon accurately, the RAPD marker was cloned and sequenced to design sequence characterized amplified region (SCAR) markers. SCAPMAR5 marker derived from pUBC411 primer yielded a single DNA band at 1061 bp. Segregation of SCAPMAR5 marker in bulk of F2 plants demonstrated that the marker was co-segregated with RAPD marker from which the SCAR marker was originated. Moreover, results of SCAR analysis in diverse melons showed SCAPMAR5 primers obtained a single 1061 bp linked to Pm-I in resistant melon PI 371795 and PMAR5. On the other hand, SCAPMAR5 failed to detect Pm-I in susceptible melons. Results of this study revealed that SCAR analysis not only confirmed melons that had been clearly scored for resistance to Pm-I evaluated by RAPD markers, but also clarified the ambiguous resistance results obtained by the RAPD markers. Key words: Cucumis melo L., Pm-I, RAPD, SCAPMAR5

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