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UNEJ e-Proceeding
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Articles 988 Documents
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Comparison of β-Glucanases Production by Acremonium sp. IMI 383068 in Batch and Continuous Culture System Jayus, Jayus
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
Publisher : UNEJ e-Proceeding

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Studies on the possible influence of physical environmental factors on extracellular b-glucanase production by Acremonium sp. IMI 383068 were undertaken, in particular to assess the role of varying agitation speed and pO2 levels, since these have been shown in other studiesto affect the yields of other fungal extracellular enzymes. The use of chemostat cultures in this present study meant that the possible influence of these two parameters on enzyme production could be controlled independently of each other and of growth rate. No other similar data on possible factors affecting fungal (1->3)- and (1->6)-b-glucanases are available in the literature, and therefore other fungal extracellular enzyme systems are used here as comparisons for this discussion. To investigate the possible relationship between enzyme production and culture morphology, the HGU values or branching frequencies of the fungal hyphae were assessed. Since substantial conidial production also occurred in these cultures of Acremonium sp. IMI 383068, it was not possible always to distinguish between mycelial branches that were true vegetative hyphae and those eventually forming the short tapered lateral conidiophores that characterize this strain. Hence HGU determinations include both of these two branch types, except where it was obvious that the branch in question was a conidiophore, adjudged by the presence of emerging conidia from it. The major findings of this study will be discused here which were mainly engaged to the relationship between production of (1->3)-b-glucanases and the growth of the fungus, where it was growth rate dependent, while that of the (1->6)-b-glucanase was growth rate independent. The production of  (1->3)-b-glucanases was shear sensitive and also depended on O2 availability in the culture medium. Based on the data from both batch and chemostat culture, it was found that there was no clear relationship between branching frequency of the fungal mycelium and b-glucanase production by the fungus.Keywords: b-glucanases, Acremonium, HGU, batch and chemostat, branching frequency
Unique Cellulase Enzyme of Bacillus firmus From Organic Waste ., Purkan; ., Sumarsih; A, Rachmadani D
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
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The objectives of this research are to isolate cellulolytic bacteria from liquid fermentation of organic waste, to determine the optimum conditions of cellulose enzyme production and characterize of crude extract cellulase enzyme activity, including pH and optimum temperature. Cellulolytic bacteria was isolated with pour plate method using specific media CMC (Carboxymethyl Celullose) and incubated at  the temperature of 370C for 24 hours. The activity of cellulolytic bacteria can be seen by the presence of transparent zone around the place where colony grows. The identification of isolates was done macroscopically, microscopically and biochemistry. Cellulolytic enzyme activity was determined with Miller method to CMC substrate. In this research, 2 isolates of cellulose producer bacteria with the biggest cellulolytic index of 1,903 (DA1) was obtained. Optimum condition of cellulose enzyme production by the bacteria was at hour-24 with the optimum molasses concentration of 0,4% and producing enzyme activity which is 0,04802 U/mL. The cellulose enzyme activity which was produced by Bacillus firmus isolate showed that the crude enzyme has  optimum activity at  pH 5 and temperature of 500C. Keywords: organic waste, Bacillus firmus,  cellulolytic bacteria, cellulose
Lipoprotein Associated Phospholipase A2 Activity and It’s Additive Value inCardiovascular Risk Stratification W, Andi; C, Miryanti; R, Saifur; H, Dadang; ., Widodo
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
Publisher : UNEJ e-Proceeding

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Background: Lipoprotein associated phospholipase A2  (Lp-PLA2) has been known as a useful  inflammatory marker of cardiovascular  risk. Framingham  score  has  also  been widely  used  in  long  term  cardiovascular  risk  stratification.  However,  it  remain  to  be clarified whether increased Lp-PLA2 activity and Framingham score significantly increase odd ratio (OR) in acute myocardial infarction (AMI) setting. We therefore investigated Lp- PLA2 activity  and  it’s  additive  value  for  Framingham  cardiovascular  risk  among myocardial infarction men.Blood samples were drawn from 60 men admitted to intensive care unit early after myocardial infarction and 40 healthy men with negative treadmill test as a control. Framingham score were calculated among all subjects. We measured Lp-PLA2 activity by ELISA. Lp-PLA2  activity was significantly higher in patients with myocardial infarction than those in control  (83.97±27.15  nmol/ml/minute  vs.  57.48±32.30  nmol/ml/minute; p=0.000). ROC analysis was performed to identify the most useful Lp-PLA2 cut-off level. Lp-PLA2 activity equal to or more than 74.21  nmol/ml/minute associated with myocardial infarction with accuracy 68%. The OR of AMI was increased by Lp-PLA2  activity added to Framingham score from 0.89(CI 95% 0.38- 2.11) to 4.48(CI 95% 1.90- 10.56) Conclusion:  Addition  of  Lp-PLA2   activity  to  Framingham  score  may  has  additive predictive value in cardiovascular risk stratification. Keywords: Lp-PLA2, myocardial infarction, cardiovascular risk stratification
Immobilization of Lipase on Surfactant-Modified Bentonite and Its Application for Biodiesel Production from Simulated Waste Cooking Oil Chrisnasari, Ruth; Yonardi, Angelina; Lie, Hesti; Widi, Restu Kartiko; Purwanto, Maria Goretti Marianti
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
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The influence of bentonite modification by cationic surfactant hexadecyl-trimethyl-ammonium bromide (HDTMA-Br) and tetramethyl ammonium hydroxide (TMAOH) on its capability to immobilize lipase was studied. Modification of bentonite was conducted by the adding of 4-6% (v/v) HDTMA-Br and TMAOH respectively. The obtained immobilized lipases then were characterized to observe the optimum pH and temperature as well as their stability during reuse application. The observed results show that there is no significant difference between the variations of HDTMA-Br concentrations to the percentage of immobilized enzyme which can immobilize lipase up to 75-78%. However, the best concentration of TMAOH is 4% (v/v) which can immobilize lipase up to 97.95%. The obtained immobilized lipases on HDTMA-Br-modified bentonite show the optimum catalytic activity on reaction temperature of 35-40 oC and pH of 7.5. In other hand, the optimum catalytic activity of immobilized lipases on TMAOH-modified bentonite is 40oC of incubation temperature and pH of 7. The immobilized lipases on both HDTMA-Br and TMAOH modified bentonite are stable enough so it could be re-used four times before its activity decreased by 48,565% and 46.83 % respectively. Keywords: Lipase, Cationic Surfactant, Bentonite, HDTMA-Br, TMAOH, Biodiesel, Immobilization.
Cloning coat protein gene of CBSD (cassava brown streak disease) at cassava (Manihotesculentum) Restanto, Didik Pudji; ., Slameto; Kriswanto, Budi; Addy, Hardian Susilo; Handoyo, Tri
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
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Cassava Brown Streak Disease (CBSD) is a major disease in cassava plants which have the serious problems in cassava plantations in the world, especially in Africa, Tanzania and India (Wassawaet., al, 2010). In Indonesia, the virus is still not optimal yet in the handling. The disease is present in plants that can destructive cassava leaves, stems and tubers.  It was greatly reduces the quality and production in the world such as India.  The decrease of cassava yield can reach 100% due to disease of CBSD (Lopez, 2003). The primer was designed from the coat protein gene of CBSD with a distance of 380 bp (Abarshiet.,al, 2012). The primers designed the forward primer (GGARCCRATGTAYAAATTTGC) and Reverse (GCWGCTTTTA  TYACAAAMGC). The RNA isolation have been used Plant Virus RNA Kit (Geneaid).  The CBSD RNA concentration around 55,2ng/ul.  The RT PCR program were one cycle of RT PCR reaction (45oC for 30 min), denaturation (45oC for 5 min) and 30 cycles for denaturation (94oC for 1 min), annealing (52oC for 30 sec), extention  (72oC for 1 min).  The results showed a single band of about 380 bp which is the  distance between the two primers were tested.  The multiplication shoot around 5 shoots per meristem explants with a combination of 0.5 ppm and 0.1 ppm BAP GA3 Keyword : Cassava Brown Streak Disease (CBSD), CASSAVA (Manihotesculentum), RT PCR
Analysis AZF Gene Deletions in Infertile Men in Indonesia Hanizar, Evi; Hinting, Aucky
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
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Infertile men with the low quantity and quality of sperm associated with gene deletions in the long arm of chromosome Y (YQ), in the region known as the AZF region (Azoospermic Factor). This study aims to analyze the AZF gene deletions in infertile men with abnormal sperm categories ranging oligozoospermia, Oligoasthenoteratozoospermia until azoospermia. This study includes the type of cross-sectional observational study and DNA samples obtained from the blood of primary infertile men. Extraction of DNA uses a DNA extraction kit and DNA amplification uses polymerase chain reaction method. The analysis includes the number, motility and morphology of sperm conducted by WHO standard. Analysis of deletion is determined from the size of the base pairs of DNA amplification product. The results showed the prevalence of deletions in azoospermic category higher than the prevalence in other categories. Sequentially, the prevalence of deletions was followed by a severe category Oligoasthenoteratozoospermia, Oligoasthenoteratozoospermia, severe oligoteratozoospermia and oligoteratozoospermia. The most frequent gene deletions are sY86 gene, followed by sY84, while the most rare gene deletions are DAZ gene / sY255. This is in contrast with previous studies because of the background sample of infertile men from several races in Indonesia, the number of samples and the location of genes analyzed in the sub-region AZF. Deletions involving many genes in the AZF subregion associated with the smaller quantity and quality of sperm. Key Words : male infertility, azoospermic factor, gene deletion, sperm abnormality
Review: Phylogenetic Similarity based on amino acid sequence and Molecular characterization of 3-Phytase from Klebsiella pneumonia ASR1 ., Sajidan; Hailu, Hailu Weldekiros
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
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Phytase is an enzyme that catalyzes the release of phosphomonester group in phytate, thereby producing lower forms of myo-inositol phosphates and inorganic phosphate.  Phytase has an important role in animal nutrition which improves the bioavailability of nutrients.  Bacteria are one of the potential sources of phytase. For this reason, more screening of phytase producing bacteria strains from environment is needed. The other sources of phytase are fungi, plants, and some animal tissues. The PhyK protein is a phytase enzyme from Klebsiella pneumonia ASR1 and expressed in Escherichia coli, which was sequenced and characterized as a 42 kDa [1], and Crystallized [2]. The objectives are to perform phylogenetic similarity and molecular evolutionary analysis of phytase and to determine active site protein motif of phytase from different sources. The sequence from PhyK was analyzed with Basic Local Alignment Search Tools (BLAST) at http://www.ncbi.nlm.nih.gov  and ClustalW Multiple Sequence analysis program (EMBL-EBI) at http://www.ebi.ac.uk/Tools/msa/clustalw2/ to get similarity of amino acid sequence and phylogenetic trees for protein.  The result shows that PhyK protein from Klebsiella pneumonia ASR1 is 91% similar with PhyK from Raoultella terrigena (emb|AJ575300.1|), 94% similar with histidine acid phosphatase family protein from Klebsiella pneumoniae 342, and 69% similar with glucose-1-phosphatase precursor Agp from Pantoea ananatis AJ13355 (BAK10365.1).The deduced amino acid sequence of the phyK gene from Klebsiella pneumonia ASR1, although containing the functional residues of histidine acid phosphatases, displayed only 31% overall homology with appA from Escherichia coli O104:H4 str. C227-11 (EGT68141.1), 30% similar with glucose-1-phosphatase from Pantoea agglomerans (gb| ABD85282.1|) and 26% to phytase from Aspergillus fumigatus (AAU93517.1). This suggests that PhyK represents a novel subfamily histidine acid phytate-degrading enzyme and is clearly distinct from other previously characterized members of this family. Keywords: Bioinformatics, BLAST, 3-Phytase, PhyK protein, Phylogenetic Similarity
Overexpression Sucrose Transporter Protein (Sut) and Sucrose Content In Genetically Modified Product (Gmp) Sugarcane(Saccharum officinarum L.) Dewanti, Parawita; Okviandari, Purnama; Oktaria, Nina; Sugiharto, Bambang
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
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Sucrose transporter (SUT) is a protein that functions to translocate sucrose from source to sink. Protein activity is determined by the accumulation of sucrose in sugarcane stem. The level of sucrose accumulation is also influenced by the activity of  sucrose phosphat synthase (SPS) and neutral invertase (NI) in conjunction with the synthesis and hydrolysis of sucrose. This aim of this study is to determine the sucrose transporter protein expression in Genetically Modified Product (GMP) of sugarcane.Genetically Modified Product GMP sugarcane events 1, 2, 3, 4, 5, 18, 20 and non GMP were used as materials. The analysis was performed by Western Blot method to observe the presence of SUT protein expression, stem sucrose content by resorcinol method to observe the translocation of sucrose, the SPS enzyme activity and NI that play a role in determining the accumulation of sucrose.The results showed that (1) an increase in SUT protein content in GMP plants events 2 and 18 with a protein band that is thicker than the non GMP plants, (2) SPS and invertase activity at all events GMP crop plants to increase compared to non GMP. Sucrose content of the stem at all events GMP plant to increase with age segment. The older age segments, the higher the sucrose  content. Keywords: sugarcane (Saccharum officinarum L.), genetically modified product(GMP), sucrose transporter protein(SUT), sucrose
Imunogenic Protein of Salivary Gland from Anopheles sundaicus Armiyanti, Yunita; Nuryady, Moh Mirza; Utomo, Sugeng Setyo; Sardjono, Teguh Wahju; Fitri, Loeki Enggar; Senjarini, Kartika
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
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Malaria is still a major problem for developing countries, including Indonesia. One approach to overcome this disease is prevention by vaccination. However, there is still no effective malaria vaccine that is applicable. The ideal malaria vaccine is a combination vaccine that can prevent the pre-erythrocytic cycle, the erythrocytic cycle and transmission process. Salivary vector-based vaccine has the potential to be developed as a malaria vaccine because it can prevent transmission process and also decrease the morbidity of the disease. Saliva from Anopheles contains vasomodulator and immunomodulatory components, that are required in the blood feeding process, but in the same time it could enhance the transmission of the malaria parasite. If the component in the salivary vector can increase pathogen infection, then vaccinating the host with its anti-substances can control the transmission of pathogens (Transmision Blocking Vaccine). Anopheles sundaicus is an important vector of malaria in coastal areas of Java, Bali, Sumatra, Kalimantan and West Nusa Tenggara islands. Repeated exposures of Salivary Gland Extract (SGE) from this vector have been proven to be able to decrease parasitemic rates in mouse model for malaria in our study. The objective of this research is to determine and localize the immunogenic protein from SGE of An. sundaicus as the first step for the characterization of its immunomodulatory component. Mosquito salivary gland protein profile of An.sundaicus was determine by SDS-PAGE. Determination of salivary glands immunogenic proteins was conducted by Western Blotting with IgG from people living from endemic area as primary antibody. Out of 15 bands appeared in SDS PAGE ranging from 24 kD to 138 kD, only two protein bands with  molecular weights of 68 and 37 kDa were the most immunogenic. Those immunogenic proteins were consistent recognized by pooled serum of people as well as by individual response. Keywords: malaria, saliva, vector, immunogenic protein, vaccine
Biochemical Resistance Mechanism of Several Genotype of Soybean to Rust Diseases Purwoko, Moh Setyo; Trisusilowati, Endang Budi; ., Amarullah
UNEJ e-Proceeding Indonesian Protein Society (IPS), International Seminar and Workshop 2014
Publisher : UNEJ e-Proceeding

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The research to identify of the content of Fenol and Flavonoid compounds which set as phytoalexin of several soybean genotype from rust diseases. Four genotypes of soybean which were examined at Unej-1, Unej-2 (moderate resistance), Burangrang (tolerance) and Lokon (susceptible). Based on rating code of IWGSR Unej-1, Unej-2 and Burangrang were proved moderate resistance and Lokon which was considered susceptible reality showed moderate susceptible. At Unej-1, Unej 2 and Burangrang which moderate resistance showed an increase of amount of Fenol and Flavonoid compounds greather than Lokon which was moderate susceptible Keywords: Biochemical Resistance, Soybean and Rust diseases

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