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Immobilization of Lactobacillus plantarum B134 Cells using Sodium Alginate for Lactose Hydrolysis in UHT Milk Lusiana Kresnawati Hartono; Tatik Khusniati; I Made Artika; Sulistiani sulistiani; Abdul Choliq
Current Biochemistry Vol. 1 No. 2 (2014)
Publisher : IPB University

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Hydrolysis of lactose in milk by β-galactosidase from immobilized bacterial cells has the potential to alleviate the problem of lactose intolerance. The present study was aimed to immobilize cells of L. plantarum strain B134 and evaluate their efficiency in hydrolyzing lactose in ultra high temperature (UHT) milk. Immobilized cells were generated by mixing cell suspensions with solutions of sodium alginate and calcium chloride. The β-galactosidase activity of the immobilized cells was tested by determining their ability in hydrolyzing lactose in UHT milk (whole milk and skimmed milk). Results showed that cells of L. plantarum strain B134 were entrapped optimally using a combination of 1 % sodium alginate, 100 mM calcium chloride and 12 % w/v cell suspension. The highest β-galactosidase activity was achieved at pH 6.5 and a temperature of 45 ºC for 5 minutes incubation time. The immobilization efficiency achieved was 28.95 %. The immobilized cells could reduce lactose by up to 85.45 % in UHT whole milk and 91.26 % in UHT skimmed milk. The times required for that reduction of lactose in UHT whole milk and UHT skimmed milk were 12 hours and 9 hours respectively. The immobilized cells could be re-used up to 4 times for efficient lactose hydrolysis for both types of milk. Therefore, immobilized cells of L. plantarum B134 have the potential to be used for lactose hydrolysis in UHT milk.
The Activity of Wungu Leaf (Graptophyllum pictum (L) Griff) Extract in Reducing Blood Glucose Level of Hyperglycemic Mice Hayatul Rahmi; I Made Artika; Norman Razief Azwar; Djarot Sasongko Hami Seno; Waras Nurcholis
Current Biochemistry Vol. 1 No. 2 (2014)
Publisher : IPB University

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Wungu leaf (Graptophyllum pictum (L.) Griff) is a plant thought to have potential use in alleviating symptoms of diabetes mellitus. The purpose of the present study was to evaluate the activity of wungu leaf extracts in decreasing blood glucose level of alloxan (200 mg/kg BW)-induced hyperglycemic mice. Extracts of wungu leaf were obtained by macerating with ethanol and then partitioning the extract with diethyl ether, ethyl acetate, and butanol. Each extract obtained was used to treat hyperglycemic mice for 28 days. The results showed that wungu leaf extracts have the ability to decrease the blood glucose level of hyperglycemic mice (dose 50 mg/kg BW). The ethyl acetate extract showed the highest activity, bringing about a decrease of blood glucose of 37.6 %. The wungu leaf extract has the potential to be developed as a source of anti-diabetic agents.
Isolation and Molecular Cloning of Cellulase Gene from Bovine Rumen Bacteria Rahadian Pratama; I Made Artika; Tetty Chaidamsari; Herti Sugiarti; Soekarno Mismana Putra
Current Biochemistry Vol. 1 No. 1 (2014)
Publisher : IPB University

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Cellulases are the enzymes that hydrolyze cellulosic biomass and are produced by the microorganisms that grow over cellulosic matters. The objective of this research was to isolate and clone cellulase gene from cellulose-degrading bacteria of bovine rumen. Cellulose-degrading bacteria was isolated from rumen fluid using a selective medium. Total RNA was isolated from selected colony having cellulose degrading activity and was used as a template for cDNA construction using reverse transcriptase polymerase chain reaction (RT-PCR) technique. The resulted cDNA was employed as a template for PCR amplification of cellulase gene using specific primers. The cellulase gene candidate obtained was cloned into the pGEM-T-Easy vector followed by determination of its nucleotide sequence. The sequence was then aligned with sequences of cellulase genes from GenBank. Results showed that a number of isolates of rumen bacteria exhibit cellulase activity and the CR-8 isolate was selected for further analysis. The successful isolation of total RNA from CR-8 was indicated by the presence of two intense bands of ribosomal RNA (23S and 16S). The reverse transcription process was successful and the amplification of cellulase gene using the specific primers F1 and R1 resulted in a DNA fragment of 1900 bp as a candidate of cellulase gene. The fragment was successfully cloned into the pGEM-T-Easy vector, and the resulted recombinant plasmid was successfully introduced into the E. coli cells. Nucleotide sequence analysis suggested that the cloned gene is cellulase gene and shares 99% homology with the endo-1,6-beta-glucanase of T. harzianum.
Trametes versicolor as Agent for Delignification of Rice Husks Laita Nurjanah; Syamsul Falah; Azmi Azhari; Suryani Suryani; I Made Artika
Current Biochemistry Vol. 1 No. 1 (2014)
Publisher : IPB University

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Rice husks contains 33.71% w/w lignocelluloses, the most abundantly available raw material on the earth for the production of biofuels and other valuable products. It is comprised of the carbohydrate polymers, cellulose, hemicellulose, and an aromatic polymer, lignin. One of the methods for removing the lignin component of rice husks is by delignification using white-rot-fungi. The aim of the study was to carry out delignification of rice husks using white-rot-fungi. The white-rot-fungi used here were Trametes versicolor and Phanerochaete chrysosporium. The study consisted of a biomass and microbial preparation, chemical assay of the rice husk, ligninase enzyme tests, and delignification of rice husks. Results showed that T. versicolor and P. chrysosporium have ligninase enzyme. The precentage of lignin from the total biomass rice husks was 23.61% w/w, and following the delignification process by T. versicolor for 20 days, the remaining lignin was 16.20% w/w, making the percentage of rice husks lignin degraded as 7.41% w/w. The biodelignification process also decreased the percentage of holocellullose, cellulose, and other extracted substances, and accordingly this increased the percentage of hemicellulose. Based on the ability of T. versicolor to degrade lignin of the rice husk at room temperature (28 ºC) as mentioned above, it can be concluded that T. versicolor has potential to be used for delignification process.
Biosorption Copper (Cu) and Mercury (Hg) by Omphalina sp. using Batch, Rotary, Biotray, and Pack Bed Flow Methods Desi Purwaningsih; I Made Artika; Tri Panji; Suharyanto Suharyanto
Current Biochemistry Vol. 3 No. 1 (2016)
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Heavy metal waste treatment often uses dangerous chemicals. Omphalina sp is a nonpathogenic fungi that can be used to reduce the harmful effects of waste treatment. The use of fungal biomass has advantages such as low operating costs, efficient, and high metal binding capacity, minimal sludge, metals can be recovered, biosorbent can be regenerated, raw materials available easily, can use inactivated microorganism, and does not require additional nutrients. In the present study optimization of the biomass utilization for waste water treatment was conducted by comparing batch, rotary, packbed flow, and biotray methods. Results showed that Omphalina sp can reduce mercury level up to 91.38% with rotary, 83.98% with biotray, 87.14% with pack bed flow, and 31.94% with batch methods respectively from initial Hg concentration of 3 ppm. Similarly, Omphalina sp can reduce copper level up to 23.58% with rotary, 22.66% with biotray, 10.53% with pack bed flow, 10.17% with batch methods respectively from initial Cu concentration of 100 ppm. Optimum absorption Hg and Cu occurs in the first one hour.
Amplification and Analysis of Cytocrome Oxidase I of Polypedates leucomystax from Bogor Agricultural University Area Perkasa Arian; I Made Artika; Syamsul Falah
Current Biochemistry Vol. 3 No. 1 (2016)
Publisher : IPB University

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DNA barcoding has become a useful tool for identifying and confirming of species within a known taxonomic framework. A large-scale effort is underway to barcode all amphibian species using the universally sequenced DNA region, a partial fragment of mitochondrial cytochrome oxidase subunit I (COI). This study was aimed to use DNA barcoding technique to identify and confirm species of Polypedates leucomystax and to analyze their phylogenetic relationship. Samples of Polypedates leucomystax were collected from Campus Area of Bogor Agricultural University. The cytochrome oxidase I gene of 600-700 nucleotides were amplified and observed in agarose gel electrophoresis. Forward sequence (604 base pairs) of COI gene was used for phylogenetic analyses. BLAST analysis against BOLD System database showed 95.75% identity with sequences of Polypedates leucomystax. The pairwise genetic distances of Polypedates leucomystax with Rhacophorus schlegelii, Limnonectes fujianensis, Fejervarya cancrivora, and Bufo melanostictus were 0.274, 0.352, 0.339, 0.339, 0.393, respectively. These results illustrated that the genetic identification is congruence with the morphological identification. Phylogenetic tree analysis showed that the samples were in one clade with other tree frogs. The DNA barcoding technique based on the sequence of COI gene can therefore be used to identify and confirm species of Polypedates leucomystax.
Deodorization of Latex Waste and Decolorization of Textile-Coloring Agent by Omphalina sp. using Batch and Continuous Methods Erna Puspasari; I Made Artika; Tri Panji
Current Biochemistry Vol. 3 No. 1 (2016)
Publisher : IPB University

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Generally industrial waste water is a pollutant to environment as it produces strong odor and color. Omphalina sp is one of white rot fungi that can be used as an odor and color effluent reducer. Omphalina sp has laccase enzyme that plays role in deodorization and decolorization. The aim of this research was to determine the best method in deodorization and decolorization among batch and continous (pack bed flow, biotray, and rotary contactor) methods. The results of deodorization and decolorization showed that the rotary contactor method was better than the batch, pack bed flow and biotray methods. At dye concentration of 50 ppm, after 24 hours treatment, the absorbance value for batch, pack bed flow, biotray, and rotary contactor methods was 0.520, 0.423, 0.425, and 0.357, respectively. At dye concentration of 150 ppm, after 24 hour treatment, the absorbance value for batch, pack bed flow, biotray, and rotary contactor methods was 0.709, 0.629, 0.658, and 0.592, respectively. At dye concentration of 50 ppm, percentage of dye absorption after 24 hour treatment for batch, pack bed flow, biotray, and rotary contactor methods was 20.550%, 35.447%, 35.141% and 45.531% respectively. At dye concentration of 150 ppm, percentage of dye absorption after 24 hour treatment for batch, pack bed flow, biotray, and rotary contactor methods was 7.320%, 17.843%, 13.987%, and 22.614%. The qmaks value at dye concentration of 50 ppm after 24 hour treatment for batch, pack bed flow, biotray, and rotary contactor methods was 0.020, 0.076, 0.083, and 0.124 respectively. qmaks value at dye concentration of 150 ppm after 24 hour treatmeny for batch, pack bed flow, biotray, and rotary contactor methods was 0.009, 0.077, 0.046, and 0.100 respectively. Odor scale for batch, pack bed flow, biotray, and rotary contactor methods decreased from 5 to 2.9, 1.4, 1.6 and 1.1 respectively.
Phylogenetic Analysis of Cytochrome Oxidase I from Buduk Toads Duttaphrynus melanostictus and Phrynoidis asper from Bogor Muhammad Dailami; I Made Artika; Mirza Dikari Kusrini; Dodi Safari
Current Biochemistry Vol. 3 No. 2 (2016)
Publisher : IPB University

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Indonesia have high diversity of Amphibians. Amphibians have an important role in ecosystem and produce many bioactive peptides. However, the genetic information of amphibians from Indonesia is very limited, especially Duttaphrynus melanostictus and Phrynoidis asper. The aims of this study are to determine the nucleotide sequence of cytochrome oxidase I (COI) from D. melanostictus and P. asper, to analyze their genetic diversity and their phylogenetic relationship. A total 668 base pairs of COI gene fragment were successfully amplified and their nucleotide sequence determined. P. asper (5 haplotypes) samples group have high haplotype diversity compared to D. melanostictus (1 haplotype). The results of Basic Local Alignment Search Tools (BLAST) to the NCBI and BOLD database, showed 99 % - 100 % identity to sequence of D. melanostictus. For the sequence of P. asper showed 99.23 % identity to sequence P. asper in BOLD database. There was no sequence of COI gene of P. asper in NCBI database. Genetic relationship among species in family Bufonidae, indicated that D. melanostictus has closer relation to P. asper than to another species, inspite of their pharapyletic characteristic. For intern species relationship of D. melanostictus, the data showed that D. melanostictus from Bogor have closer relationship to D. melanostictus from India than D. melanostictus from China.
Antibacterial Activity and Phytochemical Analysis of Geranium homeanum Turez Leaves Fri Rahmawati; Maria Bintang; I Made Artika
Current Biochemistry Vol. 4 No. 3 (2017)
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Geranium homeanum Turez is a herbaceous plant used as an empirical medicine. This research was carried out to test the antibacterial activity and determine minimum inhibition concentration (MIC) to Staphylococcus aureus and Pseudomonas aeruginosa with agar diffution method and phytochemical analysis of geranium leaves  by Harbone method. The young and old geranium leaves were blended then filtrated. The obtained filtrate was divided into two parts, one part heated by autoclave and the other was unheated. Each filtrate was tested against to Staphylococcus aureus and Pseudomonas aeruginosa. Geranium leaves filtrate having the highest antibacterial activity was heated at 500C until it  become dry powder.  The powder was used to measure MIC and phytochemical analysis.The results showed that the antibacterial activity of young leaves filtrate was higher than the old leaves filtrat, and the unheated leaves filtrate was higher than heated filtrate one. MIC of S. aureus and P. aeruginosa, were as follows 15 mg/ml and 20 mg/mL respectively. The antibacterial activity of powder geranium’s leaves  filtrate was weaker than 100 µg/mL ampicilline. Phytochemical analysis of geranium leaves showed positive contents of alkaloid and triterpenoid.
Pemanfaatan Bakteri Sebagai Pereduksi Emisi Gas Metana Pada Limbah Cair Kelapa Sawit (Elaeis guineesis Jacq.) Nisa Widya Amanda; I Made Artika; Iman Rusmana
Current Biochemistry Vol. 4 No. 2 (2017)
Publisher : IPB University

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Kelapa sawit merupakan salah satu tumbuhan penghasil minyak terbesar di Indonesia. Proses pengolahan buah kelapa sawit menjadi minyak memiliki dampak negatif, yakni terciptanya gas metana. Gas metana merupakan salah satu gas penyumbang efek rumah kaca yang menyebabkan pemanasan global. Oleh karena itu, diperlukan cara untuk menurunkan emisi gas metana. Tujuan penelitian ini adalah penggunaan bakteri untuk menurunkan emisi gas metana pada limbah cair pemrosesan buah kelapa sawit. Bakteri diisolasi dari limbah cair kelapa sawit. Terhadap isolat tunggal yang diperoleh dilakukan uji aktivitas enzim soluble methane monooxygenase (sMMO) dan particulate methane monooxygenase (pMMO). Isolat dengan aktivitas methane monooxygenase tertinggi dipilih untuk analisis sekuen DNA dan konstruksi pohon filogenetika. Tahap isolasi bakteri menghasilkan 13 isolat koloni tunggal. Uji aktivitas methane monooxygenase menunjukkan bahwa semua isolat tidak memiliki aktivitas sMMO. Sebaliknya, semua isolat memiliki aktivitas pMMO dengan tingkat berbeda yaitu dengan rentang nilai antara 0.10 – 0.22 M/mL kultur/hari. Analisis sekuen DNA menunjukkan bahwa isolat terpilih adalah Klebsiella sp. Berdasarkan uji Gram diketahui bahwa bakteri ini termasuk bakteri Gram negatif. Bakteri berpotensi digunakan untuk menurunkan emisi gas metan pada limbah cair pemrosesan buah kelapa sawit. Kata kunci: Klebsiella sp, 16S rRNA, Metana, pMMO, sMMO.
Co-Authors . SURYANI A. E. Zainal Hasan AA Sudharmawan, AA Abdillah, Ramadhani Malik Abdul Choliq ADINDA VIRGINIA DWI SETYO Agung Eru Wibowo Agung Eru Wibowo AHMAD ENDANG ZAINAL HASAN AHMAD ENDANG ZAINAL HASAN Ahmad Sulaeman Akhmad Endang Zainal Hasan Amanda, Nisa Widya AMIN FATONI Andani, Gita Putri Andita Fitri Mutiara Rizki, Andita Fitri Mutiara Antonius Padua Ratu Apipah Aprianti APON ZAENAL MUSTOPA Apon Zaenal Mustopa Arya Arendra Asri Sulfianti Azmi Azhari Azmi Azhari, Azmi Azmi, Wihda Aisarul BUGI RATNO BUDIARTO DEDI JUSADI Denny Irawati Desi Purwaningsih Dewi Sukma Dhani Luthfi Ramadhani DIMAS ANDRIANTO Djarot Sasongko Hami Seno Dodi Safari Dodi Safari Dodi Safari Dodi Safari Dwi N. Susilowati Dzihan Dinar Rabani Eliza Halim Erismar Amri Erlank Bagjavicenna Erna Puspasari Evi Nur Qolbaini Fatriani, Rizka Fina Febrianti Firda DIMAWARNITA Firdausy, Iman Akhyar Firmansyah, Ridwan Putra Fri Rahmawati Gholam, Gusnia Meilin Gita Putri Andani Gusnia Meilin Gholam H. A. E. Zainal Hasan Hani 'Athiyya Rafi Hardinsyah Harsana, Ngurah HARTUTIK EKA SUSANTI HARYANTO SUSILO Hasim - HASIM DANURI Hayatul Rahmi Herti Sugiarti Herti Sugiarti, Herti Hyakansa HANIF Ifa Manzila Iman Rusmana Inawati Inawati Irsal, Riyan Alifbi Putera Jajang Suhyana K, Popi A kurnia agustini Kurnia Agustini Laita Nurjanah Laita Nurjanah, Laita LAKSMI AMBARSARI Lasmiyanti, Metty Lusiana Kresnawati Hartono Luzicoiij, Michael Edison M. Zairin Junior M.Pd Prof. Dr. I Nyoman Sudiana . Mala Nurilmala MARIA BINTANG Meilisza, Nina Melva Louisa Mirza Dikari Kusrini MS, Yulia Atika Muhaimin Muhaimin MUHAMMAD AGUS SUPRAYUDI Muhammad Dailami, Muhammad Muhammad Nafiz Nisa Widya Amanda Noorwati Sutandyo Norman Razief Azwar Norman Razief Azwar Novik Nurhidayat Novik Nurhidayat Nuke Annisa Nasution NUNUK WIDHYASTUTI Nur Bambang Priyo Utomo Nur Hasanah Nur Hasanah Nurmala Sari Nurul Khumaida Perkasa Arian Puji Lestari Rahadian Pratama Rahmawati, Fri Rava Raisha Putra Resti Rahmawati Putri Rini Kurniasih, Rini Roedhy Poerwanto Septiany C. Palilingan Sheryn Sunni Albani Siagian, Putri Junita Siregar, Josephine Elizabeth Siti Nurjanah Soekarno Mismana Putra Soekarno Mismana Putra, Soekarno Mismana Sogandi Sogandi Sogandi Sogandi Sudarsono Suharyanto Suharyanto Sulfianti, Asri Sulistiani sulistiani Suryani Suryani Suryani Suryani Suryani Suryo Wiyono Sutoro Sutoro Syaeful Abidin Syamsul Falah Tatik Khusniati Tetty Chaidamsari Tetty Chaidamsari, Tetty Tri Panji Trini Suryani Kadir Vita Rosaline Fahri Waras Nurcholis Wasrin Syafii Wijiastuti Wijiastuti Yadi Suryadi Yahdiana Harahap Yulianto YUNITA ARIAN SANI ANWAR Zahra, Hafizh