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Isolation and Characterization of Silaffin that Catalyze Biosilica Formation from Marine Diatom Chaetoceros gracilis

HAYATI Journal of Biosciences Vol 14, No 3 (2007): September 2007
Publisher : Bogor Agricultural University, Indonesia

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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

Fatty Acid Synthesis by Indonesian Marine Diatom, Chaetoceros gracilis

HAYATI Journal of Biosciences Vol 16, No 4 (2009): December 2009
Publisher : Bogor Agricultural University, Indonesia

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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)

PEMURNIAN PROTEASE DARI BUAH DAN DAUN MENGKUDU (Morinda citrifolia L.) [Purification of Proteases from Fruits and Leaves of Noni (Morinda citrifolia L.)]

Jurnal Teknologi Dan Industri Pangan Vol 22, No 1 (2011): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

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Abstract

PEMURNIAN PROTEASE DARI BUAH DAN DAUN MENGKUDU (Morinda citrifolia L.)  [Purification of Proteases from Fruits and Leaves of Noni (Morinda citrifolia L.)] Dwi Ishartani1,2)* , Elfi2) , Nuri Andarwulan2,3) , dan Dahrul Syah2,3)   1) Program Studi Ilmu dan Teknologi Pangan, Fakultas Pertanian, Universitas Sebelas Maret 2) Departemen Ilmu dan Teknologi Pangan, Fakultas Teknologi Pertanan, Institut Pertanian Bogor 3) Southeast Asian Food and Agricultural Science and Technology (SEAFAST) Center, IPB   Diterima 15 Maret 2011 / Disetujui 24 Agustus 2011 ABSTRACT   Proteases have been widely used in cancer treatment, wounds healing, overcoming digestion disorder and other modern pharmaceutical applications. Proteases may be present in fruits and leaves of noni (Morinda citrifolia L.) since the plant has been used traditionally in wound healing. This research aimed to purify proteases from noni’s fruits and leaves at two maturity stages, i.e. fruits with green-yellow (TK2) and white-yellow (TK4) skin colour, also leaves from shoot and base. Proteases were purified through several steps consisting of extraction, precipitation using saturated ammonium sulfate and dialysis followed by electrophoresis under denaturing conditions (SDS-PAGE) and zymography. The specific activity of the four extracts showed different trend during purification. The specific activity of TK2 fruit, shoot and base leaves decreased whereas TK4 fruits increased. TK2 crude extract had a higher specific activity (3.79 U/mg) than the other crude extracts. SDS-PAGE and zymogram using 12% acrylamide indicated that the dialysates were not pure proteases. The molecular weight profiles of the TK2 dialysates were similar to TK4 dialysates, while those of shoot leaves dialysates were similar to the base leaves dialysates. The SDS-PAGE separated the enzymes in the fruit dialysates into several bands of polypeptides,i.e. 24-26kDa, 14-15kDa, 12-13kDa and the smaller ones, and also separated the enzymes in the dialysates of leaves into two bands, 70kDa and 58-61.5kDa. Protease bands detected in the zymogram of fruits were estimated at ~25kDa, ~27kDa, and 37-38kDa, whereas of those of leaves were estimated at ~29kDa and ~50kDa. The ~25kDa and ~29kDa protein bands appearing in zymogram were similar to papain’s and bromelain’s molecular weight.   Keywords: purification, proteases, Morinda citrifolia L., fruits, leaves

PENGARUH KOAGULAN DAN KONDISI KOAGULASI TERHADAP PROFIL PROTEIN CURD KEDELAI SERTA KORELASINYA TERHADAP TEKSTUR [Effect of Coagulant and Coagulation Condition to Soybean Curd Protein Profile and Its Correlation to Texture]

Jurnal Teknologi Dan Industri Pangan Vol 23, No 1 (2012): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

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Abstract

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Characteristic of Silicatein-like Protein Sponge from Nias and Lombok Marine

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

Penentuan Umur Simpan dan Pengembangan Model Diseminasi Dalam Rangka Percepatan Adopsi Teknologi Mi Jagung bagi UKM

MANAJEMEN IKM: Jurnal Manajemen Pengembangan Industri Kecil Menengah Vol 5, No 1 (2010): Manajemen IKM
Publisher : Institut Pertanian Bogor

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Abstract

Technology of corn noodle, both processed from 100% corn flour and substitution of  wheat flour (by corn flour), was developed by previous researchers but it has not been tested to be applied to the community. The objectives of this study were: (1) identify critical quality parameters and determine the shelf life of corn noodle; (2) to test the production process of dry- and wet-corn on a pilot plant scale and the small industry of wet noodles; and (3) to obtain the quantitative data of consumer acceptance from the wet noodle producers (small-medium enterprise or SMEs) and the consumers. The critical parameters of corn noodles quality during storage were the color, texture (ease of fracture), cooking loss (loss of solids due to cooking), and the degree of rancidity. Based on the aroma parameters, the shelf life of corn noodles on the storage temperature reaches 28oC was 4.6 months. Production of corn noodle routinely has been performed in a pilot plant of SEAFAST Center with a capacity 4 batch per day process. According to the wet noodles producers (small industry), the technology of the 35% substitution of corn noodles (wet noodle) can be directly adopted by SMEs without any change in the production process. The technology of 100% corn noodles can also be adopted with the addition of blanching units before sheeting process. The respondents accept the 35% substitution of corn noodles and considered no different than wheat noodles. Eighty-five percents of meatball noodles SMEs stated that are willing to use the corn wet noodle. Eighty percents of consumer’s meatball noodle said that the wet corn noodle were suitable using for meatball noodles with the acceptance value were close to the wheat noodles.   Keywords: consumer test, corn, noodles, shelf life, SMEs

Anti Cancer Activity of Chitooligomers

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

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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

Penentuan Umur Simpan dan Pengembangan Model Diseminasi dalam rangka Percepatan Adopsi Teknologi Mi Jagung bagi UKM

MANAJEMEN IKM: Jurnal Manajemen Pengembangan Industri Kecil Menengah Vol 5, No 1 (2010): Manajemen IKM
Publisher : Institut Pertanian Bogor

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Abstract

Technology of corn noodle, both processed from 100% corn flour and substitution of  wheat flour (by corn flour), was developed by previous researchers but it has not been tested to be applied to the community. The objectives of this study were: (1) identify critical quality parameters and determine the shelf life of corn noodle; (2) to test the production process of dry- and wet-corn on a pilot plant scale and the small industry of wet noodles; and (3) to obtain the quantitative data of consumer acceptance from the wet noodle producers (small-medium enterprise or SMEs) and the consumers. The critical parameters of corn noodles quality during storage were the color, texture (ease of fracture), cooking loss (loss of solids due to cooking), and the degree of rancidity. Based on the aroma parameters, the shelf life of corn noodles on the storage temperature reaches 28oC was 4.6 months. Production of corn noodle routinely has been performed in a pilot plant of SEAFAST Center with a capacity 4 batch per day process. According to the wet noodles producers (small industry), the technology of the 35% substitution of corn noodles (wet noodle) can be directly adopted by SMEs without any change in the production process. The technology of 100% corn noodles can also be adopted with the addition of blanching units before sheeting process. The respondents accept the 35% substitution of corn noodles and considered no different than wheat noodles. Eighty-five percents of meatball noodles SMEs stated that are willing to use the corn wet noodle. Eighty percents of consumer’s meatball noodle said that the wet corn noodle were suitable using for meatball noodles with the acceptance value were close to the wheat noodles.

PENGARUH KOAGULAN DAN KONDISI KOAGULASI TERHADAP PROFIL PROTEIN CURD KEDELAI SERTA KORELASINYA TERHADAP TEKSTUR [Effect of Coagulant and Coagulation Condition to Soybean Curd Protein Profile and Its Correlation to Texture]

Jurnal Teknologi dan Industri Pangan Vol 23, No 1 (2012): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

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Abstract

The research aims to study the effect of coagulation parameters to the electrophoretical profile of coagulated soybean protein as well as to texture properties and perception. Several coagulation parameters such as type of coagulant, concentration or age of coagulant, and coagulation temperature were studied. The type of coagulant used in this study were GDL (glucono delta Lactone) and tofu whey. The concentrations of GDL were 0.4%, 0.8%, and 1.2% and the ages of tofu whey were 1 day, 2 days, and 3 days. Two coagulation temperatures, 60°C and 80°C, were applied. The results of curd protein profile and texture profile analysis showed that coagulant type and coagulation condition did not affect the electrophoretical profile of soybean coagulated protein. However, the proportion of each subunit was affected significantly. Texture profiles, such as hardness, cohesiveness, and gumminess, were also affected by coagulant type and coagulation condition. Based on the subunits proportion, it can be concluded that 11 S and 7S protein proportions as well as the 11S/7S ratio correlated significantly with the hardness and gumminess of soybean curd made by tofu whey.

KRISTALINITAS DAN KEKERASAN BERAS ANALOG YANG DIHASILKAN DARI PROSES EKSTRUSI PANAS TEPUNG JAGUNG

Jurnal Teknologi dan Industri Pangan Vol 28, No 1 (2017): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

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Abstract

Rice analogues are food products made from broken rice and/or any other carbohydrate sources to have similar texture and shape to those of rice. Corn flour can be used as a material to make the rice analogues because it has various amylose contents ranging from low, medium, and high so that it is possible to make the dough with different levels of amylose contents. High temperature extrusion process may change crystallinity of starch (crystal type of A to V and degree of crystallization). Changes in starch crystallinity may influence the physical properties of the rice analogue produced. This research aimed to assess the effect of flour amylose content and temperature of extrusion on the crystallinity and its impact on physical properties of rice analogue. High amylose corn starch and corn flour were mixed at certain ratios to obtain amylose contents of 16.99, 19.35, 21.72, and 24.09%. The dough was made by adding water to the mixed flour to have a moisture content of 40%. The dough was then extruded at temperatures of 70, 80, and 90°C using a twin screw extruder (BEX-DS-2256 Berto). The observed physical properties were type of crystal, degree of crystallinity, and hardness of the rice analogue. The results showed that the increase in extrusion temperatures from 70°C to 90°C increased the degree of crystallinity of the rice analogues from 7.98-11.49% to 12.15-13.61% and hardness from 4.06-4.91 kg to 4.74-8.20 kg. The increase in amylose contents (from 16.99% to 24.09%) enhanced the hardness of the rice analogue from 4.06-4.74 kg to 4.91-8.20 kg but did not show any significant effects on the degree of crystallinity. The degree of crystallinity is positively correlated with the hardness of rice analogue (correlation coefficient r=0.8).