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Fraksinasi Peptida dari Hidrolisat Protein Ikan Selar (Selaroides leptolepis): Fractination of Peptide from Selar Fish Protein Hydrolysate (Selaroides leptolepis) Reinal Putalan; Tati Nurhayati; Ekowati Chasanah
Jurnal Pengolahan Hasil Perikanan Indonesia Vol 23 No 3 (2020): Jurnal Pengolahan Hasil Perikanan Indonesia 23(3)
Publisher : Masyarakat Pengolahan Hasil Perikanan Indonesia (MPHPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.17844/jphpi.v23i3.32202

Ikan selar (Selaroides leptolepis) merupakan ikan dengan sebaran yang cukup luas dengan produksi yang melimpah serta memiliki kandungan gizi protein yang tinggi. Peptida yang berasal dari hidrolisat protein ikan mempunyai manfaat yang besar untuk pengembangan produk pangan fungsional. Penelitian ini bertujuan mendapatkan fraksi yang memiliki aktivitas antioksidan dan inhibitor ACE dari hidrolisat protein ikan selar. Penelitian dilakukan dengan cara melakukan fraksinasi peptida menggunakan kolom kromatografi, lalu dikarakterisasi. Hasil penelitian menunjukkan bahwa fraksinasi dengan kolom kromatografi filtrasi gel mendapatkan 2 fraksi, yaitu fraksi A dan B dengan IC50 berturut-turut 4.737,95 ppm dan 529,42 ppm, serta memiliki aktivitas inhibitor ACE sebesar 90,65% dan 96.61%. Peptida fraksi B lebih potensial sebagai antioksidan dan inhibitor ACE.

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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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

PURIFICATION AND CHARACTERIZATION OF Aeromonas media KLU 11.16 CHITOSANASE ISOLATED FROM SHRIMP WASTE Ekowati Chasanah; Gintung Patantis; Dewi Seswita Zilda; Mahrus Ali; Yenny Risjani
JOURNAL OF COASTAL DEVELOPMENT Vol 15, No 1 (2011): Volume 15, Number 1, Year 2011
Publisher : JOURNAL OF COASTAL DEVELOPMENT

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Our previous study found that KLU 11.16, isolated from shrimp waste secreted chitinolytic enzymes. The crude enzyme was interesting since their chitooligosccharide was able to inhibit some pathogenic bacteria. In this study we report a purification and characterization of the chitosanase enzyme produced and the identification of the KLU 11.16. Purification of the enzyme was done two steps by ion exchange chromatography followed by gel filtration. Two out of 4 peaks from Gel Filtration step, i.e. fraction 16 and 33 were capable of hydrolyzing 100% deacetylated chitosan, indicating that both fractions contained chitosanase enzyme. The enzyme from fraction 16 had approximate molecular weight of 98.3 kDa. The enzyme worked optimally at temperature of 300C, and pH 6. Addition of Ca2+, Fe2+, K+, Na+ ions in the form of Cl2 salt and detergent Triton X-100 increased the enzyme activity, while Co2+, Mn2+ and Zn2+ ions in the same concentration decreased the enzyme acitivity. Addition of EDTA and SDS significantly decreased the enzyme activity. Molecular based identification revealed that KLU 11.16 was 99% similar to Aeromonas media.

SCREENING AND CHARACTERIZATION OF BACTERIAL CHITOSANASE FROM MARINE ENVIRONMENT Ekowati Chasanah; Dewi Seswita Zilda; Agustinus R Uria
JOURNAL OF COASTAL DEVELOPMENT Vol 12, No 2 (2009): Volume 12, Number 2, Year 2009
Publisher : JOURNAL OF COASTAL DEVELOPMENT

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Screening of extracellular chitosanase from bacterial isolates associated with marine sponges have been done. Out of 100 bacterial isolates, forty isolates were capable of forming clearing zones on the chitin media and one isolate, 34-b, produced the highest chitinolytic index. The enzymes was produced on chitin liquid medium at 37oC in a shaking waterbath for a five-day cultivation. Crude enzymes were prepared by cell-free supernatant (CFS) and concentrated through 70% (saturated) ammonium sulphate percipitation followed by dialysis. The enzymes worked best at pH and temperature of 6-7 and 60oC, respectively. The half-life (T1/2) for chitosanase activity was 500.2 min or 8.34 hours (at 37oC) and 55.12 min (at 50oC), indicating the enzyme are quite stable at that temperature. However, around 80% of the original activity was lost at 60oC after 15 min of incubation.

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

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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.

MARINE BIODISCOVERY RESEARCH IN INDONESIA : CHALLENGES AND REWARDS Ekowati Chasanah
JOURNAL OF COASTAL DEVELOPMENT Vol 12, No 1 (2008): Volume 12, Number 1, Year 2008
Publisher : JOURNAL OF COASTAL DEVELOPMENT

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Marine biodiscovery or bioprospecting activity is a search for marine products derived from marine biodiversity that can be developed for various industrial needs. Including in this activity is the process of identifying chemical compounds made by biological organisms which is often called natural product discovery. Indonesia, well known as a mega-diversity country, is one of the world hot sport of marine biodiversity. The richness of biodiversity is claimed as mirror of the richness of the chemical compounds, therefore, Indonesian waters might be rewarded with variety of chemical compounds thought to be an endless source of novel drugs and drug leads for pharmaceutical use. Up to 2007, at least 77 new compounds from 14 sponges and 19 new compounds from non-sponge organisms with pharmacological potential have been identified from Indonesian waters. To make this richness potentials becoming real in economic value, many factors should be considered. The bioactive is produced in small quantity, and the lengthy process from discovery step of a novel compound to the preclinical and clinical trials step is usually becoming a problem. Mari culture might be one among methods that can be developed in Indonesia to overcome the degradation hazard of marine resources. Conducive environment for investments, and improvement of technology on marine bioactive production through mariculture are factors to be improved to initiate and develop a sustainable biotechnology industries in Indonesia.

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

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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.

Antioxidant and Ace Inhibitor Potential of Stripe Trevally Fish (Selaroides leptolepis) Hydrolysate Reinal Putalan; Ifah Munifah; Tati Nurhayati; Ekowati Chasanah
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.v13i1.319

This study aimed to investigate the potency of fish protein hydrolysates (FPH) of stripe trevally fish (Selaroides leptolepis) as antioxidant and ACE inhibitor. The FPH was produced through enzymatically hydrolysis using protease produced by Bacillus licheniformis, a collection of Research and Development Center for Marine and Fisheries Product Processing and Biotechnology (RDCMFPPB). The FPH was fractionated using ultrafiltration membranes with molecular weight cut off (MWCO) of 10, 5 and 3 kDa. The hydrolysis degree, protein content, peptide content, antioxidant activity and Angiotensin-converting enzyme (ACE) inhibitor were observed. The result showed that FPH had maximum hydrolysis degree of 63.91% reached after 6 hours hydrolysis with protein content of 27.43 mg/mL and peptide content of 223.32 mg/mL. That FPH showed antioxidant activity (IC50) of 1941.06 ppm and ACE inhibitor of 87.82% at test concentration of 10 mg/mL. In the ultrafiltration step, the higher molecular cut off used, the higher protein content and peptide content. The lower molecular weight of the hydrolysate, the better antioxidant and ACE inhibitor activity. The best fractionation that produce potential product to be used as anti-hypertension was in mixture peptides between 3-5 kDa. The IC50 antioxidant activity was 1336.96 ppm and percentage of ACE inhibitor was 97.15% % (with a concentration of 10 mg/mL). From the results, protein hydrolysate of stripe trevally fish produced by enzymatic hydrolysis using local protease was potential as a functional ingredient, particularly as antihypertensive agent.

CYTOTOXIC ACTIVITY AND APOPTOSIS INDUCTION OF T47D CELL LINES BY Turbinaria decurrens EXTRACT Muhammad Nursid; nurrahmi dewi fajarningsih; Ekowati Chasanah
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 8, No 1 (2013): May 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.v8i1.78

Marine algae is known to contain a wide variety of biomedical compounds having pharmaceutical applications. The aim of this research was to evaluate cytotoxic activity and apoptosis induction of Turbinaria decurrens extract on T47D cell lines. Cytotoxic activity test was conducted by using MTT assay whereas detection of apoptosis was evaluated by DNA fragmentations and flow cytometry analysis. The MTT test showed that crude extract had medium cytotoxic activity to T47D, HepG2, and C28 cell lines with IC50 value of 172, againts 360 and 330 µg ml-1, respectively. After solvent partition of crude extract, the cytotoxic activity of n-hexane and ethyl acetate fractions T47D cell increased, the cytotoxic activity of n. hexane and ethyl acetate fractions T47D cell increased with IC50 value of with IC50 43.1 and 51.9 µg ml-1, respectively, whereas IC50 value of methanol fraction was 383.0 µg ml-1. Analysis of DNA fragmentation of T47D cell showed that both n-hexane and ethyl acetate fractions could not fragment DNA as a features of apoptosis. However, flow cytometry analysis by using annexin-V and propidium iodide staining revealed that n-hexane and ethyl acetate fractions could induce apoptosis in T47D cell. This research indicated that Turbinaria decurrens had potency to induce apoptosis in T47D cells.

Influence of Anthropogenic Pressures on the Bioactivity Potential of Sponges and Soft Corals in the Coral Reef Environment Hedi Indra Januar; Ekowati Chasanah; Dianne M. Tapiolas; Cherie A. Motti; Catherine H. Liptrot; Anthony D. Wright
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.v10i2.108

The wealth of marine sponges and soft corals in Indonesian waters represents a rich source of natural products. However, anthropogenic pressures potentially decrease diversity in coral reefs. Presented here are trends for tropical sponge and soft coral biodiversity and their bioactivity potential under the influence of increasing anthropogenic pressures. Samples were collected along transects (near, mid, and far) at Karimunjawa and Seribu Islands Marine National Parks and environmental parameters (salinity, pH, dissolved oxygen (DO), phosphate, nitrate, and ammonia), sponge and soft coral biodiversity, and the bioactivity potential of those organisms (50% Growth Inhibition (GI50) of cancer cell lines H460-Lung, MCF7-Breast, and SF268-CNS) are compared. The environmental conditions and biodiversity were found to be significantly different between groups of sampling sites (P0.05). Canonical Discriminant Analysis (CDA) revealed DO was the discriminant factor driving the separation between groups (90.1%). Diversity tended to be higher in the Far group with strong and significant relation to DO (R= 0.611, P0.05) and ammonia (R = -0.812, P0.05). The CDA also showed that an increase in bioactivity (low % GI50) of sponge and soft coral extracts was related to a canonical function (57.21%) consisting of high DO, high pH, and low nutrients. These findings indicate the production of bioactive compounds is related to diversity and complexity of coral reef systems. Therefore, strategies for marine protection by mitigating the impacts of anthropogenic pressures needs to be optimized in order to conserve the overall environment and sustain its natural bioactivity potential indefinitely.

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