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Isolation and Characterization of Silaffin that Catalyze Biosilica Formation from Marine Diatom Chaetoceros gracilis AGNES IMELDA MANURUNG; ALBERTA RIKA PRATIWI; DAHRUL SYAH; MAGGY THENAWIDJAJA SUHARTONO
HAYATI Journal of Biosciences Vol. 14 No. 3 (2007): September 2007
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (292.311 KB) | DOI: 10.4308/hjb.14.3.119

Abstract

The method of making silica in industries requires extreme conditions. The finding of proteins involved in the formation of biosilica from diatoms, has opened up an alternative way of production. Chaetoceros gracilis is one of the diatoms, which is potential in producing silaffin protein. This study aimed to isolate and to characterize the protein. We also analyzed the protein activity toward tetraethoxyorthosilicate (TEOS) substrate in in vitro reaction. Diatom biomass was harvested and further kept in 2% SDS/100 mM EDTA solution. Protein isolation was conducted by dissolving the silica and separating the protein by soaking in 2 M HF/8 M NH4F. Protein concentration was analyzed using Bradford method and the molecular weight was estimated through SDS-PAGE. Protein activity was observed by reacting it with TEOS substrate to form silica polymer and measured by colorimetric molibdate assay. Protein concentration was 1.20 mg/ml and appeared filamentous. The apparent molecular weights consisted of 12, 23, 42, 44 kDa. These protein was able to polymerize the silica at room temperature within 10 min. As much as 85.65 umol TEOS was polymerized per 1.4 x 106 silaffin protein per min. SEM analysis showed the formation of spherical, aggregate biosilica. Key words: Chaetoceros gracilis, silaffin protein, biosilica, polymerization
Since the primary storage nutrients in diatoms consist of lipid, they are potential for the industrial fatty acid production. High value fatty acids include arachidonic acid, eicosapentaenoic acid and docosahexaenoic acid. This study aimed to analyze fatty acid synthesis by Chaetoceros gracilis diatom during growth. There was a large increase in lipid yield from 4pg cell-1 mass of lipid per cell at the exponential phase to 283pg cell-1 at stationary phase. The lipid concentrations also increased ALBERTA RIKA PRATIWI; DAHRUL SYAH; LINAWATI HARDJITO; LILY MARIA GORETTI PANGGABEAN; MAGGY THENAWIDJAJA SUHARTONO
HAYATI Journal of Biosciences Vol. 16 No. 4 (2009): December 2009
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.16.4.151

Abstract

Since the primary storage nutrients in diatoms consist of lipid, they are potential for the industrial fatty acid production. High value fatty acids include arachidonic acid, eicosapentaenoic acid and docosahexaenoic acid. This study aimed to analyze fatty acid synthesis by Chaetoceros gracilis diatom during growth. There was a large increase in lipid yield from 4pg cell-1 mass of lipid per cell at the exponential phase to 283pg cell-1 at stationary phase. The lipid concentrations also increased significantly from the stationary phase to the death phase, but not significantly from the end exponential phase to the stationary phase. The relative percentage of saturated fatty acid (SAFA) of the total fatty acid was higher than that of monounsaturated fatty acid (MUFA) and polyunsaturated fatty acid (PUFA) at all of growth phase. The highest PUFA was found at stationary phase at the same time when SAFA was being the lowest. The majority of SAFA was palmitic acid (24.03-40.35%). MUFA contained significant proportion of oleic acid (19.6-20.9%). Oleic acid, linoleic acid and α-linolenic acid were found at every stage growth. These fatty acids are considered as precursor for production of long chain PUFA-Docosahexaenoic acid (DHA/22:6ω3) through series of desaturation and elongation step with all of desaturase enzyme (Δ8-D, Δ9-D, Δ12-D, Δ15-D, Δ17-D, Δ6-D, Δ5-D, and Δ4-D) and elongase enzyme (E).         Key words: Chaetoceros gracilis, fatty acid, synthesis, saturated fatty acid (SAFA), monounsaturated fatty acid (MUFA), polyunsaturated fatty acid (PUFA)
Anti Cancer Activity of Chitooligomers Sri Wahyuni; Fransisca Zakaria; Arief B Witarto; Dahrul Syah; Maggy T Suhartono
Jurnal Teknologi dan Industri Pangan Vol. 17 No. 1 (2006): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (532.143 KB)

Abstract

The chitin obtained from the crab industries can be used as a source for production of chitooligomers which has an important biological activity. The aims of this research was to evaluate anti cancer activity of the chitooligomers obtained from enzymatic hydrolysis using chitosanase from thermophilic bacterium Bacillus licheniformis MB2 isolated from Tompaso Manado. Media for producing the enzyme contained colloidal chitosan 1% and the enzyme was harvested after seven days of incubation at 550C. The heat stable protein enzyme was coagulated with 80% saturated ammonium sulphate and purificated using hydrophobic interaction chromatography with butyl sepharose gel. Enzyme of 0.005, 0.0085, 0.10 dan 0,17 IU/mg chitosan on soluble chitosan 1% substrate with 85% degree of deacylation were used to produce chitooligomers through incubation for one and three hours. The reaction products were analyzed (and fractionated) using HPLC. The effect of this samples on cancer cells was evaluated using K562 cells (chronic myelogenous leukemia) and investigated after being treated with MTT (3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide). In general, hydrolysates and fractionated chitooligomers showed better anti cancer activity than the 2- Bromo deoxy uridine used as positive control at similiar concentration (17 ?g/ml). Both of hydrolysates and fractionated chitooligomers (trimer to hexamer) inhibited proliferation of human K562 cancer cells line in vitro about 20.57% and 15.68% respectively.The apoptosis phenomena was found on K562 cells treated with chitooligomer hydrolysate which can be examined by Hoechts staining fluorescent method. Chitooligomers hydrolysate showed anti metastatic potential, the chitooligomers were found also as potent protease inhibitor. Keywords : chitooligomers, chitosan, anticancer
Silica, a polimerized silicon dioxide, is widely used as raw materials for food industries, such as food packaging, filter agent, biomarkers and biosensor for various analysis.  In biological sistem such as sponge, the formation of silica structure was directed by protein known as silicatein.  The aims of this research were to extract silicatein-like protein isolated from sponge live surrounding the Nias and Lombok seacost Indonesia and to study their activity to polymerize tetraethoxyorthosilic M.R.R. Lukie Trianawati; Maggy T. Suhartono; Dahrul Syah; Ekowati Chasanah
Forum Pasca Sarjana Vol. 31 No. 2 (2008): Forum Pascasarjana
Publisher : Forum Pasca Sarjana

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Abstract

Silica, a polimerized silicon dioxide, is widely used as raw materials for food industries, such as food packaging, filter agent, biomarkers and biosensor for various analysis.  In biological sistem such as sponge, the formation of silica structure was directed by protein known as silicatein.  The aims of this research were to extract silicatein-like protein isolated from sponge live surrounding the Nias and Lombok seacost Indonesia and to study their activity to polymerize tetraethoxyorthosilicate (TEOS) in-vitro.  Protein in silica spicule was isolated by collecting silica spicule, soaked in HF/NH4F buffer (pH5.0) for dissolving silica and releasing this protein.  The protein was analysed by electrophoresis SDS-PAGE to estimate the molecular weight and the concentration was analyzed by Bradford method.  The highest yield of silica spicula was 58.5% of dry weight sponge that was isolated from sponge MT37.  By SDS-PAGE, the molecular weight of protein from N6 showed three bands of 32, 27, 23 kDa, while MT5 protein was 15.5 kDa, and MT37 protein was 18 kDa.  The highest polymerization activity was 144 µmol/ml TEOS occurs at 12 hours, showed by protein isolated from sponge MT37 of Lombok Marine.   Key words: sponge, silicatein like-protein, tetraethoxyorthosilicate
Pemurnian dan Karakterisasi Kitosanase Bacillus coagulans LH 28.38 Winda Haliza; Maggy T. Suhartono; Dahrul Syah
Jurnal Penelitian Pascapanen Pertanian Vol 2, No 2 (2005): Jurnal Penelitian Pascapanen Pertanian
Publisher : Balai Besar Penelitian dan Pengembangan Pascapanen Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jpasca.v2n2.2005.76-83

Abstract

Enzim ekstrasellular kitosanase telah berhasil dimurnikan dan dikarakterisasi dari bakteri termofilik Bacillus coagulans LH 28.38 asal tanah geotermal Lahendong - Sulawesi Utara. Media pertumbuhan mengandung 0,4% koloidal kitosan sebagai sumber karbon dan diproduksi optimum pada suhu 55°C selama tujuh hari. Pemurnian enzim dilakukan dengan teknik filtrasi gel menggunakan matriks Sephadex G-IOO menghasilkan tiga puncak protein sebagai kitosanase fraksi A. B, C dan masing­ masing memiliki aktivitas sebesar 13.33 U/mg, 38.66 U/mg dan 88 U/111g. Kitosanase fraksi A memiliki suhu optimum pad a 60°C dan bekerja pad a kisaran pH luas 2-12. Kitosanase fraksi B memiliki suhu optimum 70°C dan bekerja pada kisaran pH 2-12. Kitosanase f'raksi C memiliki suhu optimum 60°C dan kisaran pH pH 2-9. Penentuan berat molekul enzim dilakukan dengan cara SDS-PAGE dan anal isis zimogram. Berm molekul enzim yang diperoleh 93, 91, 87, 77.6 dan 74 kDa. Enzim kitosanase ini dapat diaplikasikan untuk memperoleh oligomer kitosan. Oligomer kitosan sangat diminati oleh kalangan medis, karena mamiliki aktivitas anti tumor, anti kolesterol dan anti mikroba. Purification lind characterization of chitosanase from Bacillus coagulans LH 28.38Characterization and purification of the extracellular chitosanase produced by Bacillus coagulans LH 28.38 from geothermal soil origin from Lahendong - North Sulawesi has been successfully done. The enzyme was produced in medium containing 0.4% colloidal chitosan as sole carbon source and reached the highest level after 7"' day incubation at 55°C. The purification of enzyme was done by column chromatography using Sephadex G-IOO as matrix produced three peaks of protein chitosanase named as Chitosanase fraction A, Band C while has 13.3 U/mg; 38.66 U/mg and 88 U/mg activity, respectively. Chitosanase fraction A has optimal temperature at 60°C and work at range pH 2-12. Chitosanase fraction B has optimal temperature at 70°C and work at range pH 2-12. Chitosanase fraction C has optimal temperature 70°C and work at range pH 2-9. Molecular weight of chitosanase was determined by SDS-PAGE and zymograrn analysis. The molecular weight of enzyme was estimated to be 93, 91. 87. 77.6 and 74 kDa. This chitosanase could be applied to produce oligochitooligosaccaride. Oligochitooligosaccaride was used for medical popuse by medical, due to have the hipokolesterolemik, antimicrobial and antitumor activities.
Pengaruh Jenis dan Konsentrasi Koagulan Terhadap Fraksi Protein Terkoagulasi dan Tekstur Curd Kedelai (Glycine max) Dahrul Syah; Rizal Fahmi; Dadang Supriatna; RH. Fitri Faradilla
Jurnal Penelitian Pascapanen Pertanian Vol 8, No 2 (2011): Jurnal Penelitian Pascapanen Pertanian
Publisher : Balai Besar Penelitian dan Pengembangan Pascapanen Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jpasca.v8n2.2011.56-63

Abstract

Penelitian ini bertujuan mengeksplorasi pengaruh jenis koagulan pada berbagai konsentrasi terhadap fraksi protein curd yang dihasilkan serta korelasinya terhadap tekstur. Koagulan yang digunakan yaitu CaSO4. 2H2O dan CH3COOH pada konsentrasi 0,015 N, 0,03 N dan 0,045 N. Fraksi protein curd didapat dengan fraksinasi Osborne dan dianalisis lebih lanjut dengan metode sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE). Fraksi protein yang didapat dari fraksinasi Osborne yaitu albumin. globulin, prolamin, dan glutelin. Tekstur Curd kedelai dianalisis secara objektif dengan texture profile analysis (TPA) yang mencakup kekerasan, elastisitas, kohesivitas, dan gumminess. Dari hasil penelitian diketahui bahwa proses pengolahan kedelai menjadi curd menyebabkan penurunan konsentrasi fraksi protein albumin dan globulin yang signifikan. sebaliknya konsentrasi fraksi protein glutelin meningkat secara drastis. Pada konsentrasi koagulan 0.045 N. terdapat variasi pola pita fraksi protein SDS-PAGE pada kedua jenis curd(CaS04.2H20  dan CH,COOH). Jenis dan konsentrasi koagulan juga diketahui berpengaruh nyata terhadap profil tekstur objektif curd (p=0,05).Terdapat korelasi yang signifikan antara kadar fraksi protein terhadap profil tekstur objektif curd CaS04.2H2O. Sebaliknya, tekstur objektif curd CH3COOH kurang dipengaruhi oleh kadar fraksi protein. Effect of Coagulant Type and Concentration Toward Coagulated Protein Fraction and Soy Curd Texture (Glycine max] The aims of this research were to explore the effect of coagulant type and coagulant concentration on curd protein fraction. curd texture, and the correlation between these variables. Coagulants used were CaSO4. 2H2O and CH3COOH with 0.0 I5 N, 0.03 N, and 0.045 N concentration. Curd protein fractions were obtained by Osborne fractionation and was further analyzed using sodium dodecylsulfate acrylamide gel electrophoresis (SDS­ page) method. The protein obtained were albumin, globulin. prolamin, and glutelin. Curd textures that include hardness, elasticity, cohesiveness, and gumminess were analyzed by texture profile analysis (TPA). The result showed that conversing soybean to be curd caused decreasing of albumin and globulin concentration significantly. In contrast, glutelin concentration was increasing significantly. There were variations of protein fraction pattern of band SDS-PAGE between two type of curd (CaSO4. 2H2O and CH3COOH) in 0.045 N. Types and concentrations of coagulant were also influenced significantly to curd texture profile (p=0.05). There was a significant corelation between protein fraction content on objective texture profile of CaSO4. 2H2O curd. On the other hand the objective texture profile of CH3COOH curd was not influenced by protein fraction content.
Teknologi Proses Ekstrusi untuk Membuat Beras Analog (Extrusion Process Technology of Analog Rice) Faleh Setia Budi; Purwiyatno Hariyadi; Slamet Budijanto; Dahrul Syah
JURNAL PANGAN Vol. 22 No. 3 (2013): PANGAN
Publisher : Perum BULOG

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33964/jp.v22i3.114

Abstract

Tingkat konsumsi beras di Indonesia mencapai angka 139 kg/kapita/tahun, lebih tinggi dari konsumsi rata-rata di Asia Tenggara, sehingga untuk memenuhi kebutuhannya sering dilakukan impor beras. Indonesia memiliki sumber pangan lokal lain seperti jagung, sorgum, ubi kayu, ubi jalar, sagu dan lain-lain. Namun bahan pangan non beras tersebut kurang populer dibandingkan dengan beras. Oleh karena itu, perlu dikembangkan teknologi proses yang potensial untuk mengolah bahan pangan lokal non beras menjadi beras analog. Teknologi ekstrusi merupakan salah satu teknologi yang telah digunakan untuk pembuatan beras analog dengan bahan baku beras patah. Belakangan, teknologi ini juga mulai digunakan untuk pembuatan beras analog dari bahan pangan non beras. Karakteristik beras analog yang mirip dengan beras alami dapat dicapai dengan mengontrol parameter-parameter kritis ekstrusi seperti karakteristik dan komposisi bahan, suhu ekstrusi, kecepatan ulir dan sebagainya. Studi menunjukkan bahwa beras analog bisa dibuat dari bahan pangan non beras. Keberhasilan teknologi ini juga akan memperluas peluang fortifikasi dengan menggunakan beras analog sebagai pembawa zat gizi. seperti protein, vitamin dan mineral, sesuai dengan tujuannya. Makalah ini mengkaji hasil-hasil penelitian pembuatan beras analog dengan teknologi ekstrusi baik dengan menggunakan bahan beras patah maupun bahan non beras yang disertai dengan dan tanpa fortifikasi.Indonesia rice consumption level is very high and up to 139 kg/capita/year, higher than that of average consumption level in South East Asia, so that import of rice is frequently needed to fill the need of population. Indonesia is actually rich in local food sources other than rice; such as corn, sorghum, cassava, sago, etc. but they are not as popular as rice. Therefore technology for the production of analog rice using the localbased non-rice food sources is needed. Extrusion technology has been used to produce analog rice from broken rice as its raw material. Recently; extrusion technology has also been used to develop analog rice using non-rice food material. The characteristic of analog rice which is similar with the natural rice could be achieved by controlling the critical extrusion parameters, such as the characteristics and composition of raw material, the temperature of extrusion, the speed of screw etc. The success of the analog rice production from the non rice food material will open up opportunities for fortification program using analog rice as a carrier for the nutrient target. This paper reviews the research reports for analog rice production with extrusion technology using variety of raw materials; including broken rice and the non-rice food material, with and without fortification.