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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 518 Documents

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Effect of Probiotic Lactobacillus sp. Dad13 on Humoral Immune Response of Balb/C Mice Infected with Salmonella typhimurium Ika Dyah Kusumawati; Eni Harmayani; Widya Asmara
Indonesian Journal of Biotechnology Vol 11, No 1 (2006)
Publisher : Universitas Gadjah Mada

An indigenous strain of lactic acid bacterium (LAB) identified as Lactobacillus spp. Dad13 (Dad13), isolated from traditional fermented buffalo milk, was found to be potential as probiotic. The aim of this research was to study the effect of probiotic Dad13 on humoral immune response of Balb/C mice infected with Salmonella typhimurium. The specific objective was to find out the effect of different Dad13 consumption time (before and along with infection of S. typhimurium) on the humoral immune response of Balb/C mice. The experiment was conducted by in vivo trial on 20 males of Balb/C mice, age of 6-8 weeks, fed with AIN-93 standard diet. The mice were assigned into 4 groups. Each group received the following treatments, ie. Dad13 only, Dad13 before infection, Dad13 along with infection and Salmonella infection only. A volume of 100 µl Dad13 cell suspensions (1010 CFU/ml) were given by oral forced feeding daily for a week, at week 3 for group before infection and at week 4 for group of Dad13 only and Dad13 along with infection. Salmonella infection (109 CFU/ml) was given once orally at week 4 to all groups except group treated with Dad13 only. The humoral immune response of Balb/C mice was detected 2 weeks after infection by measuring the titers of IgG and IgA specific from serum and mucosal intestinal liquid samples using Enzyme-linked Immunosorbent Assay (ELISA) method. The result indicated that humoral immune response of Balb/C mice consuming Dad13 before and along with Salmonella infection were significantly different (p<0.05). Dad13 consumption along with Salmonella infection increased circulated IgG and IgA as well as secretory IgA. It can be concluded that Dad13 probiotic feeding along with infection increased humoral immune response more significantly compared to that before infection.

In Vitro Screening of Falcataria moluccana (Miq.) with Gall Rust (Uromycladium tepperianum (Sacc.) Filtrate as Media Selection Asri Insiana Putri; Mohammad Na'iem; Sapto Indrioko; Sri Rahayu; Ari Indrianto
Indonesian Journal of Biotechnology Vol 19, No 2 (2014)
Publisher : Universitas Gadjah Mada

In vitro screening of Falcataria moluccana (Miq.) was conducted by tissue culture method. Seeds fromtwo different site of community forest, 400 m (S1) and 800 m (S2) above sea level, were used as material.Double concentration of MS (Murashige & Skoog, 1962) with 40 mg/l gall rust (Uromycladium tepperianum(Sacc.) fi ltrate were used for media selection. The results of this research showed that 66 % axenic plantlets invitro from S1 and 27 % from S2 were still survived after 3 months incubation without subculture. The meanof fresh weight (2. 21 ± 0. 26 g) and dry weight (1. 97 ± 0. 12 g) from S1 plantlets lower than the mean of freshweight (2. 87 ± 0,18 g) and dry weight (2. 16 ± 0. 14 g) from S2 plantlets. Qualitative of terpenes, saponins andquantitative of total phenolics were analyzed from those gall rust extract, as source of fi ltrate media, attackedand un-attacked of F. moluccana. They all qualitatively have capability to produce terpenoid and saponin. Itis notice that U. tepperianum, un-cultured pathogen, contain of those compound that may play a role as codeterminantsof pathogenecity. While the highest total phenolic compound were contained in gall rust extract(2. 35 %), followed by attacked F. moluccana branches (1. 18 %) and un-attacked F. moluccana branches (0. 44%). This indicated that phenolic compound in gall rust has higher activity as a response of F. moluccana to U.tepperianum pathogen pressures and result of this study suggest the great value of gall rust fi ltrate for use asmedia selection in vitro.

Cloning and Expression of hGAD65 Gene in E. Coli BL21 Rista Nikmatu Rohmah; Soraya Widyasari; A. Aulanni’am; F. Fatchiyah
Indonesian Journal of Biotechnology Vol 18, No 1 (2013)
Publisher : Universitas Gadjah Mada

The aim of this study is to construct the hGAD65 gene and to identify the hGAD65 clone by using PCR & RFLP. The samples were derived from normal person & DM patient’s blood. Blood DNA was isolated by salting out method and then amplified by PCR with a pair of specific primer, GAD65-F-BamH1-807 & GAD65-R-Xho1-945. The PCR-product was cloned into vector pET-28a and the pET28a-hGAD65-clone was transformed into E.coli BL21 competent cells. The pET28a-hGAD65-clone was confirmed by PCR and RFLP by BamH1 & XhoI. The PCR product of pET28a-hGAD65-clone was one band of 159bp and has two bands 5.3 kb and 159 bp by RFLPwith both restriction enzymes. The GAD65 protein is expressed in 65kD of pET28a-hGAD65-clone. PET28a-hGAD65-clone was able to recognize by gold standard monoclonal antibody specifically. These results indicated that the hGAD65 gene inserted into pET28a properly and provided the GAD65 protein expression. Key words: hGAD65, PCR, pET-28a, RFLP

Distribution of Camphor Monooxygenase Genes in Soil Bacteria N. Ngadiman; Hikaru Suenaga; Masatoshi Goto; Kensuke Furukawa
Indonesian Journal of Biotechnology Vol 10, No 2 (2005)
Publisher : Universitas Gadjah Mada

In microbial degradation of camphor, the first step is oxidation by multiunit enzyme, camphor monooxygenase, encoded by cam genes (camA,B,C). Seven camphor-utilizing bacterial strains have been isolated from soil at various locations. CamA,B,C genes of Pseudomonas putida strain PpG1 and strain GF2001 were used as probes to explore their abundance in the camphor-utilizing bacteria. Southern analysis revealed that all of the cam genes of GF2001 could hybridize well to the SpeI-digested genomic DNA of strains tested, whereas PpG1 cam genes were not. This result suggested that the GF2001 type cam genes are widely distributed among the camphor- utilizing strains in the environment. Thus strain GF2001 and seven newly isolated strains share a common evolutionary origin.

The effect of methanol extract of soybean seeds (Glycine max L.Merr.) on the histology and immunohistochemical distribution of Cyp19 aromatase in rat testis (Rattus norvegicus L.) Retno Aryani; Sukarti Moeljopawiro; Laurentius Hartanto Nugroho; Pudji Astuti
Indonesian Journal of Biotechnology Vol 20, No 2 (2015)
Publisher : Universitas Gadjah Mada

Soybean (Glycine max L. Merr.) contains phytoestrogens that have a chemical structure resembling estrogen in the body. They function like estrogen and antiestrogen, affecting the metabolism of sex steroid hormones. This research aimed to determine the effect of the methanol extract of soybean on the histological structure and distribution of immunohistochemical Cyp19 aromatase in rat testis . Twenty males of Wistar rats were divided into 4 groups of 5. The frst group was the control and the second to fourth groups were given soybean extract (250 mg/kg of body weight, 500 mg/kg of body weight) and genistein (0.3 mg/kg of body weight), respectively, for 52 days. The results of this study indicate that the effect of methanol extract from soybean caused weight gain, and the weight of the testis and epididymis decreased. In addition, the histological results showed that seminiferous tubules were reduced in size, became irregular, were separated by a wide interstitium, and spermatogenic cells were decreased. The immunohistochemical results showed that the expression of Cyp19 aromatase in the rats decreased both in spermatocyte cells and Leydig cells. It could be concluded that the methanol extract of soybean induced testicular damage and reduced Cyp19 aromatase expression in rat testis.

The Detection of Mutational Changes in Sorghum using RAPD T. Taryono; Paramita Cahyaningrum
Indonesian Journal of Biotechnology Vol 16, No 1 (2011)
Publisher : Universitas Gadjah Mada

Sorghum is a multifunction plant due to its high economic value as a source of food, feed and industrial raw material of biofuel. Sorghum improvement can be done through mutation breeding and this research was conducted to evaluate the power of mutation breeding by observing the difference between mutant and its original counterpart. Varieties of sorghum Durra, Zhengzu and their mutants B-100 and Zh-30 were arranged in completely randomized design. DNA extraction was done using a modified CTAB method. After purification, quantification, and dilution, PCR was carried out for RAPD analysis. The result showed that Durra and Zhengzu varieties were significantly different in plant height, number of leaves and seed colour, however the mutant and its original counterpart cannot be differentiated morphologically. RAPD can be used to differentiate mutant and its original counterpart by observing the specific band pattern from each primer.

Simultaneous clustering analysis with molecular docking in network pharmacology for type 2 antidiabetic compounds Nur Azizah Komara Rifai; Farit Mochamad Afendi; I Made Sumertajaya
Indonesian Journal of Biotechnology Vol 22, No 1 (2017)
Publisher : Universitas Gadjah Mada

The database of drug compounds and human proteins plays a very important role in identifying the protein target and the compound in drug discovery. Recently, a network pharmacology approach was established by updating the research paradigm from the current “one disease-one target-one drug” to a new “drug-target-disease network”. Ligand-protein interactions can be analyzed quantitatively using simultaneous clustering and molecular docking. The docking method offers the ability to quickly and cheaply predict the ligand-protein binding free energy (DG) in structure-based virtual screening. Meanwhile, simultaneous clustering was used to find subgroups of compounds that exhibit a high correlation with subgroups of target proteins. This study is focused on the interaction between the 306 compounds from medicinal plants (brotowali Tinospora crispa, ginger Zingiber officinale, pare Momordica charantia, sembung Blumea balsamifera, synthetic drugs (FDA-approved) and the 21 significant human proteins associated with type 2 diabetes. We found that brotowali (B018), sembung (S031), pare (P231), and ginger (J036, J033) were close to the synthetic drugs and can possibly be developed as antidiabetic drug candidates. Likewise, the proteins AKT1, WFS1, APOE, EP300, PTH, GCG, and UBC which assemble each other and which have a high association with INS can be seen as target proteins that play a role in type 2 diabetes.

Identification of Pediococcus Strains Isolated from Feces of Indonesian Infants With in vitro Capability to Consume Prebiotic Inulin and to Adhere on Mucus W. Widodo; Nosa Septiana Anindita; Tiyas Tono Taufiq; Tutik Dwi Wahyuningsih
Indonesian Journal of Biotechnology Vol 17, No 2 (2012)
Publisher : Universitas Gadjah Mada

The aim of this experiment was to identify isolates obtained from feces of Indonesian infants and to evaluate their capability as probiotics. Identification of isolates was carried out based on morphology, physiology and biochemical identifications, and molecular identification based on 16S rRNA sequence. Morphological and physiological identification was carried out based on Gram staining, shape, motility, spore formation and catalase production. Biochemical identifications based on production of CO2 and NH3 from glucose, the ability to grow on different temperature (10 and 45°C) and pH (4.4 and 9.6), and different salt concentration (6.5 and 18%). Probiotics capability of isolates was assayed on the ability to grow on low pH (pH 2.0), on different bile salts concentration (0.3; 0.5; 1.0 and 1.5%), the capacity to grow on media with inulin as the only carbon source, and in vitro adhesion ability on porcine mucin. Morphological, physiological and biochemical identification suggest that all of isolates belong to lactic acid bacteria. Further molecular identification of five isolates showedthat isolates AA, BE and BK were strains of Pediococcus acidilactici (similarity 99%), while isolate AP and AG were strains of Lactobacillus casei (similarity 99-100%). Probiotic assays showed that more than 80% of cells of Pediococcus acidilactici isolates AA, BE and BK were viable after grown on pH 2.0 for 90 min, and around 80% of cells from the same isolates were survived on media supplemented with bile salt 1.5% for 2 h. All of isolates had high adhesion capacity as seen by more than 75% of cells attached on pig gastric mucin. Investigationof isolates to grow on inulin showed Pediococcus acidilactici isolate BE was able to consume inulin as the only carbon source. It is concluded that Pediococcus acidilactici isolate BE was a candidate probiotics and subject to further in vivo evaluation using animal models to examine their beneficial health effects.Key word : Pediococcus acidilactici, Lactobacillus casei, human origin and probiotics.

Histological Features of Catecholaminergic Neuron in Substantia Nigra Induced by Paraquat Dichloride (1,1-dimethyl-4,4 bipyridinium) in Wistar Rat as A Model of Parkinson Disease Tri Wahyu Pangestiningsih; Woro Danur Wendo; Yulfia Nelymalik Selan; Filphin Adolfin Amalo; Nemay Anggadewi Ndaong; Victor Lenda
Indonesian Journal of Biotechnology Vol 19, No 1 (2014)
Publisher : Universitas Gadjah Mada

Paraquat dichloride has been used by farmers as a herbicide to kill the grass. On the other hand, paraquatdichloride is harmful if enters to the body, causing Parkinson’s disease, since it is disrupting dopamineproduction in the substantia nigra pars compacta or dopamine pathways Nigro striatal pathway. The studywas done to fi nd out the histological changes of catecholaminergic neurons and Nigro striatal pathway causedby paraquat dichloride treatment in Wistar rats as a model of Parkinson’s disease.Twenty-two Wistar rats 3,5 months old were divided into 4 groups, 5 rats each. Group I (control group)were injected with aquabidest, while groups II, III, and IV were injected intraperitoneally with paraquatdichloride in aquabidest, with the dosage 5 , 10 and 15 mg/kg bw respectively. The rats were injected onceper week for 6 weeks. Three days after the last injection, the rats were anesthetized using xylasin (2 mg/kg)and ketamine (20 mg/kg) intramuscularly, and then were intracardiac perfused using physiological saline asprerinse solution, followed by 10% buffered formalin solution as a fi xative. After animals were fi xed, the brainswere removed and embedded in paraffi n block and cut in 12 μm thickness for immunohistochemistry stainingusing tyrosine hydroxylase antibody. The results of staining then were observed under light microscope andanalyzed descriptively.The results showed that the catecholaminergic neurons were distributed in the substantia nigrapars compacta in all treatment groups, however, the cell density were found decreased only in group IV.Catecholaminergic neurons appear in the bipolar and multipolar form, while dopamine ‘Nigro striatal pathway’was found exist in all treatment groups. From our study, histologycally the decreased of catecholaminergicneurons is only found in rats that received paraquat dichloride in dose 15 mg/kg bw for 6 weeks.

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.

Page 16 of 52 | Total Record : 518

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