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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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Abstract

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. 
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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Abstract

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.
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

Abstract

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.
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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Abstract

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. 
Fatty Acid Profile, Carotenoid Content, and In Vitro Anticancer Activity of Karimunjawa and Lampung Sea Cucumber Ekowati Chasanah; Yusro Nuri Fawzya; Kustiariyah Tarman; Hedi Indra Januar; Muhammad Nursid
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 11, No 3 (2016): December 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.v11i3.269

Abstract

Fatty acids and carotenoid has been known as an anticancer agent on both preventing and treating cancer disease. This study was conducted to analyze the fatty acid profile, carotenoid and in vitro anticancer activity of 12 sea cucumber harvested from Karimunjawa and Lampung waters. The aim of the study was to determin the potency of sea cucumbers as raw material for nutraceutical products. Fatty acid profile and carotenoid content were characterized by gas chromatography and spectrophotometry techniques, while in vitro anticancer activity was assessed by MTT assay against cervix (HeLa), breast (T47D and MCF-7) and colon (WiDR) cancer cells. Results of the study showed polyunsaturated fatty acid (PUFA) dominated the composition of fatty acids in the samples from both locations. Holothuria sp. was detected to contain the highest amount of carotenoid. Furthermore, the highest in vitro anticancer activity was detected also in the sample of Holothuria sp. The activity of 30 ppm Holothuria sp. extract against HeLa cell was detected to be almost equal to the 5 ppm doxorubicin control. Concentration of 5 ppm Holothuria sp. extract also showed positive result in killing 50% of MCF-7 and T47D, but capable to 100% kill HeLa and WiDR cells. At concentration of 25 ppm, the extract was able to kill all the 4 cells tested. Statistical analysis showed the amount of carotenoid and two particular fatty acid compounds (docosadienoic and eicosapentaenoic acid) significantly (P0.05) contributed to the cytotoxic activity that was found in the sea cucumber samples. Those compounds were found in highest concentration from Holothuria sp harvested from Lampung waters, thus being the most prospective raw material for nutraceutical or functional food ingredient with anticancer potency.
SCREENING AND CHARACTERIZATION OF L-GLUTAMINASE PRODUCED BY BACTERIA ISOLATED FROM SANGIHE TALAUD SEA Ekowati Chasanah; Usman Sumo Friend Tambunan; Tanti Yulianti
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.v7i3.6

Abstract

L-glutaminase (L-glutamine amidohydrolase, EC 3.5.1.2) is a very important enzyme due to its role as flavor enhancer and antileukemic agent. Salt-tolerant L-glutaminase produced bymarine bacteria is favorable in food industries. This study describes the screening of L- glutaminase producing marine bacteria from Sangihe-Talaud Sea, North Sulawesi, Indonesia.Screening of L-glutaminase was performed using a liquid medium and identification of selected isolate was  performed using molecular-based 16S rDNA. Results showed that there were 7 isolates produced positive results of L-glutaminase, and one of them (II.1 isolate) produced the highest activity, i.e 147.99 U/L, equivalent to the specific activity of 62.32 U/mg. The isolate then selected for further study. Bacterial identification based on 16S rRNA sequencing has revealed that the isolate was 96% similar to Pseudomonas aeruginosa strain CG-T8. Characterization of extracellular L-glutaminase from the II.1 isolate showed that the enzyme worked optimally at temperature  of 37-45 °C and pH 7. The enzyme was stable when NaCl solution was added up to 8% and  began to decrease on addition of NaCl solution of 16% and 20% with relative activity of 79% and 74%, respectively. The effect of metal ions, e.g Mn2+, Mg2+, and Co2+ in the form of chloride salt, were able to increase enzyme activity, whereas the addition of other metal ions (Zn2+, Fe3+, and Ca2+) decreased the activity. The molecular weights of L-glutaminase was estimated around42 kDa and 145 kDa.
BIOETHANOL PRODUCTION FROM SEAWEED PROCESSING WASTE BY SIMULTANEOUS SACCHARIFICATION AND FERMENTATION (SSF) Andi Hakim; Ekowati Chasanah; Uju Uju; Joko Santoso
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 12, No 2 (2017): August 2017
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.v12i2.281

Abstract

Seaweed processing waste has been used for bioethanol production through simultaneous saccharification and fermentation (SSF). SSF is commonly used for bioethanol production to shorten the process and to increase the yield of ethanol produced by Trichoderma reesei and Saccharomyces cerevisiae. The aim of this research was to obtain the best concentration of T. reesei and S. cerevisiae to produce bioethanol by SSF. The concentration of T. reesei and S. cerevisiae used was 0 (control), 5, 10, 15 and 20% (v/v). The SSF process was carried out by using shaking incubator at 35 °C and rotation of 150 rpm for 3 days. The untreated and hot water treated seaweed processing waste used in this study have moisture content values of 12.94±0.08% and 15.38±0.19%, ash content values of 16.72±0.08% and 18.39±0.19%, lignin content values of 15.38±0.11% and 12.74±0.38%, and cellulose content values of 26.92±0.57% and 34.57±0.81%, respectively. The result of SSF process of seaweed processing waste showed that different concentrations of T. reesei and S. cerevisiae (control, 5, 10, 15 and 20%) yielded significant effect (p0.05) on the total reducing sugars and ethanol produced. The Duncan Multiple Range Test (DMRT) showed that the treatment 10% of T. reesei and S. cerevisiae concentration in the seaweed processing waste treated with hot water was the best treatment producing highest yield of ethanol.
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
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

Abstract

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.
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

Abstract

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.