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Pemanfaatan Limbah Pengolahan Agar sebagai Komponen Medium Produksi Selulase dari Mikroba Yusro Nuri Fawzya; Amelia Latifa; Nita Noriko
Jurnal Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 9, No 1 (2014): Juni 2014
Publisher : Balai Besar Riset Pengolahan Produk dan Bioteknologi Kelautan dan Perikanan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/jpbkp.v9i1.99

Abstract

Pengolahan agar dari rumput laut berkembang pesat di Indonesia. Hal ini berkaitan dengan produksi rumput laut yang cenderung meningkat dari tahun ke tahun. Produksi yang meningkat ini diikuti dengan meningkatnya limbah pengolahan agar. Limbah ini diketahui memiliki kandungan selulosa yang tinggi. Pemanfaatan limbah ini umumnya untuk pembuatan pupuk dan komponen pakan. Penelitian ini bertujuan untuk memanfaatkan limbah pengolahan agar sebagai komponen medium produksi enzim selulase dari mikroba. Tiga jenis isolat mikroba yaitu PMP1206, Serratia marcescens SGS 1609, dan isolat bakteri PC3, dikultivasi dalam medium padat yang mengandung karboksimetil selulosa (CMC). Isolat mikroba yang menghasilkan zona bening paling besar, dipilih dan enzim diproduksi dalam medium cair yang mengandung selulosa dari limbah agar. Enzim yang dihasilkan dari perlakuan terbaik dilakukan karakterisasi. Medium produksi enzim adalah medium sintetik minimal (MSM) cair dengan penambahan 1% limbah agar yang telah diberi perlakuan dengan NaOH 0, 2, 4, dan 6%. Inkubasi dilakukan pada suhu 30 oC, 150 rpm. Pengambilan sampel dilakukan setiap hari untuk diuji aktivitas enzimnya. Hasil seleksi isolat menunjukkan bahwa S. marcescens SGS 1609 menghasilkan zona bening paling besar dengan indeks selulolitik 2,25. Produksi enzim selulase dari isolat ini pada medium limbah agar menunjukkan bahwa waktu optimal produksi enzim diperoleh pada inkubasi selama 1–3 hari dari perlakuan NaOH 6%. Enzim yang dihasilkan bekerja optimum pada pH 6 dan suhu 50 oC. Enzim bersifat stabil terhadap panas. Pada suhu 40–60 oC selama 4 jam penurunan aktivitas enzim tidak lebih dari 30%. Aktivitas selulase meningkat dengan penambahan ion Ca2+, dan Mg2+, dan menurun dengan adanya 10 mM ion Zn2+.
Kitosan Oligosakarida: Produksi dan Potensinya sebagai Antibakteri Ariyanti Suhita Dewi; Yusro Nuri Fawzya
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 1, No 1 (2006): December 2006
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v1i1.75

Abstract

Kitosan
Cloning of a Transglutaminase Gene from Streptomyces thioluteus TTA 02 SDS 14 Seprianto Seprianto; Dewi Seswita Zilda; Yusro Nuri Fawzya; Suharsono Suharsono; Puspita Lisdiyanti; Agustinus Robert Uria
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 11, No 1 (2016): May 2016
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v11i1.189

Abstract

Microbial Transglutaminase (MTGase, EC 2.3.2.13) is an enzyme that catalyzes the transfer of acyl group. Many microbial strains that produce MTGase belong to Streptomyces members. This research was aimed at cloning of a MTGase gene. PCR–based screening of ten MTGase-producing streptomyces isolates from soil in West Nusa Tenggara led to detection of one potential isolate, designated as TTA 02 SDS 14. The partial  MTGase-encoding gene (470 bp)  was amplified by PCR and sequenced. The sequence result indicate its similarity of 93 % with that of Streptomyces cinnamoneus. The 16S rRNA gene analysis showed its identity as Streptomyces thioleteus. Fosmid-based construction of a genomic library from the isolate  and subsequent screening led to the isolation of  a ~40-Kb fosmid harboring a MTGase gene.
Identification of SGS 1609 Cellulolytic Bacteria Isolated from Sargassum spec. and Characterization of The Cellulase Produced Yusro Nuri Fawzya; Stenny Putri; Nita Noriko; Gintung Patantis
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 8, No 2 (2013): August 2013
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v8i2.87

Abstract

Bacterial isolate from seaweed designated as SGS 1609 was previously found to be able to produce cellulase represented by formation of clear zone on solid medium containing carboxymethylcellulose (CMC). This research was conducted to identify the isolate and determine optimum production time as well as characterize the cellulase produced. The isolate was identified using  16s-rRNA gene analysis. Cellulase production was conducted by cultivating the isolate in the liquid medium containing CMC followed by centrifuging to get supernatant as the crude enzyme. The enzyme was then concentrated using ammonium sulfate precipitation and ultra filtration. The concentrated enzyme having higher activity produced from the concentration process was then characterized  to determine its optimum pH and temperature, heat stabilization, metal ions effect and substrate specificity. The result showed that the SGS 1609 isolate was identified as Serratia marcescens with 99%  similarity. The isolate produced cellulase optimally at 4 days incubation. Ultra filtration produced higher enzyme activity compared to NH4-sulfate precipitation. The enzyme concentrated by ultra filtration worked optimally at the  pH of 7, temperature of 50 oC, stable at the temperature of 60 oC for 240 minutes and was increased its activity by Ca2+ and Mg2+ ions. On the other hand, the enzyme was inhibited by Fe3+, Zn2+ and Na+ ions, but was not relatively affected by K+ and EDTA. The use of conventional agar producer waste  treated with 6% NaOH gave highest activity compared to other substrates.
SCREENING OF THERMOSTABLE PROTEASE PRODUCING MICROORGANISMS ISOLATED FROM INDONESIAN HOTSPRING Dewi Seswita Zilda; Eni Harmayani; Jaka Widada; Widya Asmara; Hari Eko Irianto; Gintung Patantis; Yusro Nuri Fawzya
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 7, No 3 (2012): December 2012
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v7i3.5

Abstract

Although many proteases had been studied and characterized, only a few of them are commercially available.  Protease thermostability is one of the crucial properties for industrialapplication. This research aimed to isolate and to screen the potential isolate which produce thermostable protease. There were 6 isolates (BII-1, BII-2, BII-3, BII-4, BII-6 and LII), isolated using solid Minimal Synthetic Medium (MSM) supplemented with 1.5% skim milk, that have, protease activity. Based on the 16S-rRNA gene sequencing analysis, isolates BII-1, BII-2 and BII- 6 were identified as Bacillus licheniformis, isolates BII-3 and BII-4 were identified as Bacillus subtilis, while isolate LII was identified as Brevibacillus thermoruber. Three isolates (BII-6, BII-4 and LII) were then further investigated for the second screening step using liquid MSM supplemented with 1% skim milk. The isolates (BII-6, BII-4 and LII) optimally produced protease when they were cultivated at 35, 30 and 50o C respectively after 22 h of incubation. Protease produced by BII-6, BII-4 and LII had optimum temperature  of 65, 60 and 85o C, optimum pH at 7-8, 8 and 9 and stable up to 100 min at 55, 60 and 75o C respectively.
OPTIMIZATION OF Bacillus sp. K29-14 CHITINASE PRODUCTION USING MARINE CRUSTACEAN WASTE Agustinus Robert Uria; Ekowati Chasanah; Yusro Nuri Fawzya
JOURNAL OF COASTAL DEVELOPMENT Vol 8, No 3 (2005): Volume 8, Number 3, Year 2005
Publisher : JOURNAL OF COASTAL DEVELOPMENT

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (227.735 KB)

Abstract

Chitin is present in large quantities in the marine crustacean waste disposed by seafood processing industries, making it very desirable as the substrate for producing chitinase, a hydrolytic enzyme of considerable interest in many industrial and agricultural applications. In our work, crustacean waste powder and its combination with colloidal chitin at different concentrations (0.5, 1.0, and 1.5%) were utilized to optimize the chitinase production by the bacterium, Bacillus sp. K29-14. The results showed that the chitinase production with the three different substrate concentrations was relatively constant in the range of 0.2 to 0.3 U/ml during 12 days cultivation, although there was a bit reduction after day 8. This activity profile seems to be similar to that of the protein content. Whereas the chitinase production on the media containing crustacean waste powder and its combination with colloidal chitin at the three concentrations showed that the highest activity (3.0 to 4.6 U/ml) was achieved on day 7 and 8. The specific chitinase activity with the waste powder at different concentrations of substrate (0.5, 1.0 and 1.5%) was increasing slowly during a nine-day cultivation. The optimal chitinase production (4.6 U/ml) was achieved with the combined substrate of 0.5% on day 8.
NOVEL MOLECULAR METHODS FOR DISCOVERY AND ENGINEERING OF BIOCATALYSTS FROM UNCULTURED MARINE MICROORGANISMS Agustinus Robert Uria; Yusro Nuri Fawzya; Ekowati Chasanah
JOURNAL OF COASTAL DEVELOPMENT Vol 8, No 2 (2005): Volume 8, Number 2, Year 2005
Publisher : JOURNAL OF COASTAL DEVELOPMENT

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (489.976 KB)

Abstract

Metagenomics is a powerful cultivation-independent approach, which can be applied to gain access to the biocatalysts from uncultured marine microorganisms. Discovery of marine biocatalysts by this approach, in general, involves four main steps. First, a metagenomic library containing a pool of biocatalyst-encoding genes is constructed from a marine environment, which can be done by various methods, including cloning of enzymatically-digested DNA, uncut DNA, and PCR-amplified products. Second, the metagenomic library is screened for the genes of interest by employing the activity assay of expression product, in situ  hybridization, or Polymerase Chain Reaction (PCR). Third, the obtained target genes, both functional and phylogenetic genes, are sequenced and analysed by using bioinformatic tools in order to gain information on the functional and structural properties as well as the microbial sources of the encoded biocatalysts. Finally, the target genes are expressed in suitable microbial hosts, thereby producing the corresponding recombinant biocatalysts. All existing methods in engineering of marine biocatalysts for the performance improvement can be classified into two main strategies: (i) rational design and (ii) directed evolution. Rational design, which may include the use of resctriction enzyme(s) and splicing by overlap extension (SOE), requires information on the biocatalyst`s structural and functional properties to alter specific amino acid(s). Whereas directed evolution, including error-prone PCR technique and gene shuffling, needs no such information.
Teknik Peningkatan Performa Enzim Asal Mikroba Laut yang Tidak dapat dikultur Agustinus Robert Uria; Dewi Seswita Zilda; Yusro Nuri Fawzya
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 1, No 1 (2006): December 2006
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v1i1.72

Abstract

Tingginya keanekaragaman dan kelimpahan mikroba laut menunjukkan potensinya sebagai sumber paling menjanjikan untuk penemuan produk alami yang bernilai industri.  Di antara berbagai bentuk produk alami, enzim termasuk paling diminati dalam dunia industri baik dalam industri kimia, farmasi maupun makanan.  Hal ini berkenaan dengan peranannya sebagai katalis yang ramah lingkungan, ekonomis dan bersih (Wahler Reymond, 2011).
Bacterial Diversity of a Microbial Mat from Hot Spring at Wartawan Beach, Lampung and Its Potential as a Source of Hydrogenases Gintung Patantis; Ekowati Chasanah; Yusro Nuri Fawzya; He Pe Qing; Zhang Xue Lei
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 13, No 1 (2018): May 2018
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v13i1.323

Abstract

Biohydrogen produced from thermophilic hydrogenases is an ideal and clean energy sources. As the biggest tectonic area in the world, Indonesia is potential for thermophile isolation. The aims of this study were to analyze the bacterial diversity of a microbial mat from hot spring at Wartawan beach, Lampung and to analyze the potency of microbial mat for hydrogenases, using clone library method. The diversity of 16S rRNA showed that the microbial mat sample contained 9 phyla of bacteria, and dominated by Cyanobacteria and Proteobacteria. These phyla indicate that the bacterial community of the microbial mat consisted of phototrophic and heterotrophic groups. In addition, a microbial mat of Wartawan beach environment might be influenced by marine environment and hydrothermal vent which was indicated by detection of both associated bacteria. The diversity of hydrogenase genes using NiFe hydrogenase (NiFe) and FeFe hydrogenase (FeFe) genes showed that Cyanobacteria was specifically related to NiFe, while Firmicutes was associated with FeFe. Proteobacteria and Bacteroidetes, however, were detected for both genes. The detected hydrogenase genes indicate that the microbial mat from hot spring at Wartawan beach is a promising source for hydrogenases isolation and further applications for biohydrogen production as a renewable energy. 
Chemical Pretreatment and Enzymatic Saccharification of Seaweed Solid Wastes Pujoyuwono Martosuyono; Andi Hakim; Yusro Nuri Fawzya
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 10, No 2 (2015): August 2015
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v10i2.130

Abstract

The technical feasibility of seaweed waste utilization as a resource of renewable energy was investigated in this paper. The production of fermentable sugars from seaweed waste was studied by dilute sulfuric acid and sodium hydroxide pretreatment and further enzymatic hydrolysis. Pretreatment was conducted by using 1.0 and 2.0% dilute sulfuric acid w/v and 4 and 5% sodium hydroxide w/v for 30 min at 121 oC. Pretreated seaweed wastes were analyzed by X­Ray Diffraction (XRD) to examine the crystallinity index of the cellulose and observed using Scanning Electron Microscopy (SEM) to examine the changes in structure of cellulose fiber. Saccharification of pretreated seaweed waste was carried out using crude cellulase enzyme provided by Pulp and Paper Research Center in Bandung. Saccharification was done in shake flask with 20% of substrate in citrate phosphate buffer at 30 oC and 50 oC, agitation of 150 rpm in shaking incubator for 48 h. Samples were collected at 2, 6, 12, 24 and 48 h for further analysis. Enzyme concentrations were varied between 10­50 U/g dry samples. The results showed that dilute acid and base pretreatment of seaweed solid waste can be used to improve the digestibility of seaweed waste. It successfully acted by reducing the lignin content and degrading the structure of cellulose from crystalline into amorphous form which is more susceptible to the enzyme action.The optimum pretreatment condition was shown by 4% NaOH at 121 oC for 30 min, producing the most fermentable sugar concentration. Sugar concentration produced by saccharification was optimum at 50 oC, enzyme concentration of 50 U/g sample for 24 h base pretreatment. The results of the experiment were expected to contribute in the process development of bioconversion of lignocellulosic materials into renewable energy sources.