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Isolasi Dan Penapisan Aktinomisetes Laut Penghasil Antimikroba Sunaryanto, Rofiq; Marwoto, Bambang; Irawadi, Tun Tedja; Hartoto, Liesbetini
ILMU KELAUTAN: Indonesian Journal of Marine Sciences Vol 14, No 2 (2009): Jurnal Ilmu Kelautan
Publisher : Marine Science Department Diponegoro University

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Abstract

 Telah diiakukan isolasi dan penapisan aktinomisetes laut yang mampu menghasilkan senyawa antimikroba. Isolasi diiakukan ditiga tempat berbeda yaitu di Pantai Barat Banten, Pantai Utara Cirebon, dan Pantai Selatan Yogyakarta. Isolasi dilakukan dengan dua metode pre-treatment yaitu dengan metode pengasaman dan metode pemanasan. Dari hasil isolasi diperolehjumlah total isolat sebanyak 50 isolat. Setelah diiakukan penapisan diperoleh 4 isolat yang mampu menghambat Eschereschia coli, 5 isolat mampu menghambat Streptococcus aereus, 4 isolat mampu menghambat Bacillus subtilis, 4 isolat mampu menghambat Pseudomonas aeroginosa, 5 isolat mampu menghambat Candida albican, dan 4 isolat mampu menghambatAspergillus niger. Hasil identifikasi morfologi dan DNA dari salah satu isolat yang memiliki aktivitas antibakteri paling kuat (isolat A11) adalah Streptomyces sp. Secara morfologi isolat A11 memiliki hifa yang bercabang dengan kantong spora pada ujung hifa. Minimum Inhibition Concentration (MIC) isolat A11 terhadap Bacillus subtilis sebesar 120,86 μg/ml.Kata kunci: Isolasi, penapisan, aktinomisetes laut, antimikrobaIsolation and screening of antimicrobial-producing marine actinomycetes has been conducted on isolates taken from West Banten, North Cirebon, and South Yogyakarta Coasts. Two methods pretreatments were applied i.e. acid and heat shock method. The research 50 isolates. The screening reavealed four isolates which has ability to inhibi Eschereschia coli, 5 isolates could inhibited Streptococcus aereus, 4 isolates could inhibited Bacillus subtilis, 4 isolates could inhibited Pseudomonas aeroginosa, 5 isolats could inhibited Candida albican, and 4 isolatescould inhibited Aspergillus niger. Result of  identification morphology and DNA of isolate A11 it’s Streptomyces sp. Morphology of isolate A11 haves branching hyphae with spore sack at the end of hyphae. The Minimum Inhibition Concentration (MIC) of isolate A11 to Bacillus subtilis was 120,86 μg /ml.Key words: Isolation, Screening, Marine Actinomycetes, Antimicrobial.
Isolation and Characterization of Antimicrobial Substance from Marine Streptomyces sp. SUNARYANTO, ROFIQ; MARWOTO, BAMBANG; IRAWADI, TUN TEDJA; HARTOTO, LIESBETINI
Microbiology Indonesia Vol 4, No 2 (2010): August 2010
Publisher : Indonesian Society for microbiology

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Abstract

Isolation and purification of antimicrobial active substance produced by marine Actinomycetes has been carried out.  Marine sediment samples were obtained from six different places at Banten West Coast.  Isolation was conducted using two pretreatment methods,  acid and heat shock pre-treatment.  A total of 29 Actinomycetes isolates were obtained from the various sediment samples collected, then tested for antimicrobial activity against Escherichia coli ATCC 25922, Staphylococcus aureus ATCC25923, Pseudomonas aeruginosa ATCC27853, Bacillus subtilis ATCC 66923, Candida albicans BIOMCC00122, and Aspergillus niger BIOMCC00134.  Among the isolates, isolate A11 was the most activity to Gram-positive and Gram-negative bacteria, and morphological observation and identification using 16S rRNA showed that the isolate was similar to Streptomyces sp.  Production of active compound from A11 isolate used yeast peptone medium.  Purification of active compounds was carried out using silica-gel-column chromatography and preparative HPLC.  A single peak of active compounds was detected by HPLC, which showed a retention time of 8.35 min and maximum absorbance in UV visible at 210 nm and 274.5 nm respectively.
Cyclo (Tyrosyl-Prolyl) Produced by Streptomyces sp.: Bioactivity and Molecular Structure Elucidation SUNARYANTO, ROFIQ; MARWOTO, BAMBANG; HARTOTO, LIESBETINI; IRAWADI, TUN TEDJA
Microbiology Indonesia Vol 5, No 2 (2011): June 2011
Publisher : Indonesian Society for microbiology

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Abstract

Determination of bioactivity by minimum inhibitory concentration (MIC) methods and molecular structure identification of antibiotic produced by Streptomyces sp. have been carried out. The antibiotic was produced by liquid culture using Streptomyces sp. isolate. Purification of antibiotic was carried out by silica gel column chromatography and preparative HPLC. Molecular structure identification was carried out using ESI-MS, 1H NMR, 13C NMR, and 13C DEPT NMR. Pure antibiotic showed inhibition activity to Gram-negative and Gram-positive bacteria. MIC to Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC27853 , Staphylococcus aureus ATCC25923, and Bacillus subtilis ATCC66923 were 27.0, 68.7, 80.2, and 73.7 μg mL-1 , respectively. Identification using ESI-MS showed that the molecular weight of this antibiotic was 260 g mol-1 , and molecular formula was C14H16N2O3 . Elucidation of molecular structure using 1HNMR, 13C NMR, and 13C DEPT NMRshowed that antibiotic was cyclo(tyrosyl-prolyl).
Enzymatic and Acid Hydrolysis of Sago Starch for Preparation of Ethanol Production SUNARYANTO, ROFIQ; HANDAYANI, BERTI HARIASIH; SAFITRI, RATU
Microbiology Indonesia Vol 7, No 2 (2013): June 2013
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.5454/mi.7.2.68

Abstract

A series of studies on the hydrolysis of Sago starch for ethanol fermentation had been conducted. Hydrolysis of sago starch was carried out using sulfuric acid 2.5% and amylase(s) enzymes. The concentrations of sago starch used in this experiment were  5, 10, 15, 20, and 30% (w/v). The highest hydrolyzate  containing reducing sugar was used as substrate for ethanol fermentation by Saccharomyces cerevisiae FNCC 3012. The results indicated  that hydrolysis using 2.5% sulfuric acid for 120 min at 121 °C produced 6.6%  (w/v)  reducing sugar and hydrolysis using α-amylase and Dextrozyme DX produced more reducing sugar, 7% (w/v) and 17.1% (w/v), respectively. The fermentation of hydrolyzed sago starch by S. cerevisiae FNCC 3012 produced ethanol 7.98% (v/v).
ISOLASI, IDENTIFIKASI, DAN KARAKTERISASI BAKTERI ASAM LAKTAT DARI DADIH SUSU KERBAU sunaryanto, rofiq; marwoto, bambang
Jurnal Sains dan Teknologi Indonesia Vol 14, No 3 (2012)
Publisher : Jurnal Sains dan Teknologi Indonesia

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Dadih is one of the traditional fermented milk products of West Sumatera. Dadih contain a lot of lactic acid bacteria that acts as a coagulant and preservative. Some lactic acid bacteria also act as a probiotic agent because of characteristics that are resistant to acidic conditions. Some of the main requirements of microbes that can be used as probiotic microbes were resistant to low pH, bile salts, able to colonize, and having antimicrobial activity. Each species of the genus Lactobacillus havedifferent characteristics. This characteristic are influenced by the environment in which the bacteria live. Isolation, identification and characterization of lactic acid bacteria derived from buffalo milk were done. The results of isolated lactic acid bacteria was Lactobacillus plantarum. The characterization of Lactobacillus plantarum indicated that these isolates were able to survive in a concentration of medium containing 0.5% bile salts, resistant to acidic media until pH 2, have antimicrobial activity (inhibit Escherichia coli, Staphylococcus aureus and Enterococcus faecalis).
Optimizing Fermentation Medium to Produce Cyclosporin a Using Response Surface Methodology Sunaryanto, Rofiq
Makara Journal of Technology Vol 16, No 1 (2012)
Publisher : Directorate of Research and Community Services, Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (276.349 KB) | DOI: 10.7454/mst.v16i1.235

Abstract

Response surface methodology (RSM) is a three factorial model which illustrates the relationship between one or more independent variables. RSM can be used to optimize the fermentation medium for the production of Cyclosporin A from the isolate Tolypocladium inflatum. The optimal point of the response surface area is predicted by using a second-degree polynomial model and applying the statistic model obtained from the central composite design (CCD). The results of optimizing the fermentation medium for Cyclopsorin using the three independent variables of glucose, casein, and KH2PO4 show that all three of the independent variables affect the production of Cyclosporin A. There is a positive interaction between the independent variables of glucose and casein, however, there is no visible interaction between glucose with KH2PO4  and casein with KH2PO4. By using the mathematical model the total optimum result obtained is 1230.5 mg L-1, glucose concentrate 28.5 g L-1, KH2PO4  concentrate 0.74 gL-1, and casein concentrate 9.8 g L-1. Laboratory validation shows that Cyclosporin A productivity is 1197.285 mg L-1.  There is a value difference of 2.7% between the expected productivity of Cyclosporin A using the mathematical model and the actual production in laboratory tests.
PENGARUH KOMBINASI BAKTERI ASAM LAKTAT TERHADAP PERUBAHAN KARAKTERISTIK NUTRISI SUSU KERBAU Sunaryanto, Rofiq
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol 4, No 1 (2017): June 2017
Publisher : Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29122/jbbi.v4i1.2064

Abstract

Effects of Lactic Acid Bacteria Combination on Characteristic Change of Buffalo Milk Nutrition Determination of physical and chemical characteristics on fermented milk using a mixture of Lactobacillus bulgaricus, Streptococcus thermophilus, and Bifidobacterium sp. has been conducted. Fermentation was carried out for 20 hours at 37ºC in a facultative aerobic condition. Physical characteristics were performed by comparing the physical properties before and after fermentation such as viscosity and texture, including chemical properties such as pH, acidity, protein, fat, and sugar contents. Inoculants of lactic acid bacteria were varied using a combination of lactic acid bacteria mixture. Results showed that the combination of bacteria inoculants used has no significant effect on changes in protein, fat, ash, and water contents. However, the effects on total acid, pH, sugar content, and physical appearance such as viscosity and texture were significant. The combination of L. bulgaricus, S. thermophilus and Bifidobacterium sp. (A6) produced a soft texture appearance and high viscosity. The combination of inoculant bacteria L. bulgaricus with S. thermophilus (A5) produced a texture similar to A6 but with a lower viscosity than A6.Keywords: Lactobacillus bulgaricus, Streptococcus thermophilus, Bifidobacterium sp, buffalo milk, fermentation. ABSTRAKPenelitian untuk mengetahui karakteristik fisik dan kimia hasil fermentasi susu kerbau menggunakan campuran Lactobacillus bulgaricus, Streptococcus thermophilus, dan Bifidobacterium sp telah dilakukan. Fermentasi dilakukan selama 20 jam pada suhu 37ºC secara aerobe fakultatif. Karakteristik fisika dilakukan dengan membandingkan sifat fisik seperti kekentalan dan tekstur, serta sifat kimia yang meliputi kandungan protein, lemak, gula, pH, keasaman sebelum dan sesudah fermentasi. Inokulan bakteri asam laktat divariasikan menggunakan kombinasi campuran bakteri asam laktat. Hasil penelitian menunjukkan bahwa kombinasi inokulan yang digunakan dalam proses fermentasi susu kerbau tidak berpengaruh nyata terhadap perubahan kadar protein, lemak, abu, dan air. Namun demikian berpengaruh nyata terhadap total asam, pH, kadar gula, dan penampakan fisik seperti kekentalan dan tekstur. Kombinasi inokulan L. bulgaricus, S. thermophilus dan Bifidobacterium sp. (A6) menghasilkan susu fermentasi dengan penampakan tekstur halus dan kekentalan yang lebih padat. Kombinasi bakteri L. bulgaricus dengan  S. thermophilus (A5) menghasilkan tekstur yang mirip dengan A6, namun demikian menghasilkan kekentalan yang lebih rendah dibandingkan A6.Kata kunci: Lactobacillus bulgaricus, Streptococcus thermophilus, Bifidobacterium sp, susu  kerbau, fermentasi
OPTIMALISASI MEDIA PRODUKSI AMILOGLUKOSIDASE MENGGUNAKAN FERMENTASI MEDIA PADAT Sunaryanto, Rofiq; Marasabessy, Ahmad
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol 3, No 1 (2016): June 2016
Publisher : Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29122/jbbi.v3i1.19

Abstract

Amiloglukosidase merupakan salah satu enzim yang banyak digunakan dalam industri gula cair. Dalam proses fermentasi, komposisi medium fermentasi sangat berpengaruh terhadap aktivitas amiloglukosidase. Optimalisasi media padat proses produksi amiloglukosidase secara fermentasi padat dengan menggunakan isolat Aspergillus niger telah dilakukan. Optimalisasi media padat dilakukan dengan menentukan rasio dedak:pati terbaik yang selanjutnya dikombinasikan dengan beberapa sumber nitrogen. Dalam penelitian ini sumber nitrogen yang digunakan adalah sumber nitogen organik dan sumber nitrogen anorganik. Sumber nitrogen organik yang digunakan antara lain Corn Step Liquor dan pepton, adapun sumber nitrogen anorganik yang digunakan adalah amonium nitrat, amonium sulfat, dan amonium pospat. Fermentasi dilakukan selama 120 jam pada suhu 30ºC.Hasil percobaan menunjukkan bahwa kombinasi perbandingan media sumber karbon dedak:pati 1:1 menghasilkan produktivitas amiloglukosidase tertinggi 724 Unit/ml. Amonium sulfat merupakan sumber nitrogen anorganik terbaik dan mampu menghasilkan produktivitas amiloglukosidase sebesar 823 Unit/ml, sedangkan Corn Step Liquor (CSL) yang merupakan sumber nitrogen organik mampu menghasilkan produktivitas amiloglukosidase lebih tinggi dibandingkan pepton dengan produktivitas sebesar 884 Unit/ml.Keywords: Aspergillus niger, amyloglucosidase, fermentation, carbon source, nitrogen source  ABSTRAKAmiloglukosidase adalah salah satu enzim yang banyak digunakan dalam industri gula cair. Dalam proses fermentasi, komposisi medium fermentasi sangat berpengaruh terhadap aktivitas amiloglukosidase. Pada penelitian ini telah dilakukan optimalisasi media padat pada proses produksi amiloglukosidase secara fermentasi padat dengan menggunakan isolat Aspergillus niger. Optimalisasi media padat dilakukan dengan menentukan rasio dedak:pati terbaik yang selanjutnya dikombinasikan dengan beberapa sumber nitrogen. Sumber nitrogen yang digunakan adalah sumber nitrogen organik dan anorganik. Sumber nitrogen organik yang digunakan antara lain adalah Corn Step Liquor (CSL) dan pepton, adapun sumber nitrogen anorganik yang digunakan adalah amonium nitrat, amonium sulfat, dan amonium fosfat. Fermentasi dilakukan selama 120 jam pada suhu 30ºC. Hasil percobaan menunjukkan bahwa kombinasi perbandingan media sumber karbon dedak:pati 1:1 menghasilkan produktivitas amiloglukosidase tertinggi, yaitu 724 unit/mL. Amonium sulfat merupakan sumber nitrogen anorganik terbaik dan mampu menghasilkan produktivitas amiloglukosidase sebesar 823 unit/mL, sedangkan CSL menghasilkan produktivitas amiloglukosidase lebih tinggi dibandingkan pepton, yakni sebesar 884 unit/mL.Kata kunci: Aspergillus niger, amiloglukosidase, fermentasi, sumber karbon, sumber nitrogen
UJI KEMAMPUAN Lactobacillus casei SEBAGAI AGENSIA PROBIOTIK Sunaryanto, Rofiq; Martius, Efrida; Marwoto, Bambang
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol 1, No 1 (2014): December 2014
Publisher : Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29122/jbbi.v1i1.546

Abstract

Growth Ability of Jatropha Curcas L. Explants on the In Vitro Media Containing IBA and BAProbiotic product is one of the applications of biotechnology that utilize lactic acid bacteria, especially lactobacilli. Some important requirements for microbes that can be used as probiotic include resistance to low pH, ability to grow on bile salts and colonize, and having antimicrobial activity. Each species of the genus Lactobacillus has different characteristics. These characteristics are strongly influenced by the environment in which the bacteria live. This study was carried out in order to characterize Lactobacillus casei which was isolated from dadih. The result of the experiment showed that the isolated L. casei was able to grow on the bile salt at the concentration of 15%, resistant to acid media until pH 2, had antimicrobial activity (significantly inhibited the growth of Escherichia coli, Staphylococcus aureus, and Enterococcus faecalis). The local L. casei isolate has a potential application for use as probiotic microbe.Keywords: Lactobacillus casei, probiotic, lactic acid bacteria, characterization,dadih ABSTRAKProduk probiotik merupakan salah satu aplikasi bioteknologi yang memanfaatkan bakteri asam laktat terutama jenis Lactobacillus. Beberapa syarat utama mikroba yang dapat difungsikan sebagai mikroba probiotik antara lain tahan terhadap pH rendah, mampu tumbuh pada garam empedu, mampu berkoloni, memiliki aktivitas antimikroba. Masing-masing spesies dari Genus Lactobacillus memiliki karakteristik yang berbeda-beda. Karakteristik ini sangat dipengaruhi oleh lingkungan dimana bakteri tersebut hidup. Pada penelitian ini telah dilakukan karakterisasi Lactobacillus casei yang merupakan hasil isolasi dari susu kerbau fermentasi. Dari hasil percobaan menunjukkan bahwa L.casei hasil isolasi mampu hidup sampai dengan konsentrasi garam empedu 15%, tahan terhadap media asam sampai dengan pH 2, memiliki aktivitas antimikroba (positif menghambat Escherichia coli, Staphylococcus aureus, dan Enterococcus faecalis). L. casei yang merupakan isolat lokal memiliki karakteristik yang berpotensi untuk digunakan sebagai mikroba probiotik.Kata kunci: Lactobacillus casei, probiotik, bakteri asam laktat, karakterisasi, dadih
Isolation, Identification, and Evaluation of Antimicrobial Activity of Active Compound Produced by Marine Actinomycetes isolate A32 Sunaryanto, Rofiq; Marwanta, Edy
ANNALES BOGORIENSES Vol 19, No 2 (2015): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1111.48 KB) | DOI: 10.14203/ab.v19i2.216

Abstract

Isolation, identification, and evaluation of antimicrobial activity of active compound produced by marine actinomycetes isolate A32 has been conducted. Production of active compound using isolate A32 was conducted by glucose, yeast, peptone medium. The fermentation was carried out at 30ºC for 5 days. The broth of supernatant was extracted using ethyl acetate. Purification of active compound used column chromatography and eluted stepwise with the chloroform and methanol solvent. Antimicrobial activity was monitored by the agar diffusion, and microbial test used as followed Escherichia coli ATCC 25922, Staphylococcus aureus ATCC25923, Bacillus subtilis ATCC 66923, and Candida albican BIOMCC00122. Results of isolation and purification of active compound produced by isolate A32 showed that this compound has a molecular weight of 503.1 g/mol with molecular formula C26H37N3O7. Analysis of spectrum using 1HNMR and COSY, this compound was suspected as Madumycin II. The antibacterial activity assay showed that this active compound was able to inhibit to the growth of Staphylococcus aureus ATCC25923 and Candida albican BIOMCC00122.