Abstract. Patantis G, Dewi AS, Fawzya YN, Nursid M. 2019. Identification of Beche-de-mers from Indonesia by molecular approach. Biodiversitas 20: 537-543. Sea cucumbers have been commercially traded around the world due to their nutritional values and health benefits. Sea cucumbers are marketed as processed sea cucumber, also known as beche de mer, teripang, haisen or gamat. Due to the high global demand of beche-de-mers, there have been several cases of fraud due to the visual shape and color similarities of processed sea cucumbers. Additionally, the local name diversity of Indonesian sea cucumbers making the identification of beche-de-mers difficult. Therefore, this study aimed to identify commercial sea cucumbers from selected locations in Indonesia by molecular approach. The targeted Cytochrome c oxidase subunit I (COI) was used in this research. A total of 25 beche-de-mers was purchased from local markets in Indonesia, i.e. Boalemo (17 local names), Pesawaran (5), Surabaya (2) and West Lombok (1). Results showed that 22 species of beche-de-mer were successfully identified, ie. Acaudina sp., Actinopyga echinites, A. lecanora, A. miliaris, Bohadschia argus, B. marmorata, B. ocellata, B. vitiensis, Holothuria sp. H. atra, H. coluber, H. edulis, H. excellens, H. fuscocinerea, H. fuscogilva, H. fuscopunctata, H. scabra, Pearsonothuria graeffei, Phyrella sp., Stichopus herrmanni, S. horrens and S. monotuberculatus. The identification of Indonesian beche-de-mers using molecular method could support the identification of the local sea cucumbers to overcome mislabeled and fraud. Correct identification is important for the record of beche-de-mer trade in Indonesia and further conservation management.

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Indonesia has a great sea cucumber resource which is useful as functional food. Even though much information about the biopotency of sea cucumbers have been published, however, studies on nutrition value of Indonesian sea cucumbers are still beneficial due to the great diversity and uniqueness of each species. The present work was intended to obtain information about chemical composition and fatty acid profile of 4 (four) types of sea cucumbers which are frequently found in Halmahera water, North Maluku (Molucca), one of sea cucumbers producing area in Indonesia. The samples coded as H-03, H-04, H-05 and H-10, were identified as Bohadschia argus, Holothuria fuscogilva, Thelenota ananas, and Actinophyga lecanora. All sea cucumbers showed high protein content, more than 60% (drybase/db), except for T. ananas which had the lowest protein content (48.26% db). As for fats, T. ananas showed the highest amount (2.35%db) with dominated saturated fatty acids (SFA). On the other side, A. lecanora showed higher value of PUFA compared to SFA and MUFA. Palmitic acid (C16:0) was the most abundant SFA in the most samples with the amount of 0.49-4.9 mg/g sample. Among PUFA, eicosapentaenoic acid/EPA (C20:5n3) was detected, and eicosatrienoic acid (C20:3n6) was relatively higher than the others. The ratio of 6/3 fatty acid was 1.2-3.2 showing that total 6 fatty acid was higher than total 3 fatty acid. However, this value is safe enough to protect against chronic and degenerative diseases.

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.

Isolasi dan karakterisasi parsial kolagen dari teripang gamma (Stichopus variegatus) telah dilakukan. Isolasi dilakukan melalui tiga tahap yaitu preparasi, ekstraksi dan isolasi. Tahap preparasi meliputi tahapan penyiangan, pencucian dan perendaman dalam akuades, dalam alkohol, dalam larutan Tris-HCl dan etilenadiaminatetraasetat  (EDTA) dan perendaman dalam larutan natrium hidroksida (NaOH). Ekstraksi dilakukan dengan perendaman asam asetat 0,5M. Isolasi dilakukan dengan cara pengendapan menggunakan NaCl dilanjutkan proses dialisis. Isolat kolagen teripang gamma yang diperoleh memiliki rendemen sebesar 16,40% (bobot kering), nilai pH 6,08 dan derajat putih 77,02%. Gugus fungsi kolagen terdiri dari amida A (3412 cm-1), B (2929 cm-1), I (1654 cm-1), II (1554 cm-1), dan III (1239 cm-1). Asam amino utama penyusun kolagen yaitu glisin, prolin, dan alanin, masing-masing sebesar 16,88%; 6,71%; dan 6,42%. Kolagen yang dihasilkan merupakan kolagen tipe I, diduga terdiri dari 3 rantai a1 yang homolog dengan berat molekul 130,33 kDa.

Thermostability is among of the vital enzyme characteristics for industrial application. Brevibacillus thermoruber LII was obtained as a potential isolate from the previous research which screened the potential thermostable protease producing bacteria from Indonesian hotspring. The newly thermostable protease produced by thermophilic Brevibacillus thermoruber LII had been purified and characterized. It was predicted that the pure enzyme obtained from Brevibacillus thermoruber LII was homo hexameric, having molecular weight of 36 kDa unit protein and its native was 215 kDa. In addition, it was also a neutral metalo serine protease according to biochemical tests that it was totaly inhibited by PMSF (Phenylmethanesulfonyl fluoride) and EDTA (Ethylenediaminetetraacetic acid). It showed optimum activity at pH of 8 and active in acidic buffer (up to pH of 4). All of metal ion in the form of chloride salt (2.5 mM) which were tested on the enzyme enhanced the enzyme activity but Li Keywords: thermostable protease, Brevibacillus thermoruber, hotspring ABSTRAK Ketahanan terhadap panas merupakan salah satu sifat enzim yang penting untuk aplikasi industri. Brevibacillus themoruber LII diperoleh dari penelitian sebelumnya pada penapisan isolat potensial penghasil protease tahan panas dari sumber air panas Indonesia. Protease baru tahan panas yang dihasilkan oleh Brevibacillus thermoruber LII sudah dimurnikan dan dikarakterisasi. Diprediksi bahwa enzim murni yang dihasilkan oleh mikroorganisma tahan panas, Brevibacillus thermoruber LII merupakan heksamer dengan berat molekul unit yang sama yaitu 36 kDa dan nativenya 215 kDa.Lebih lanjut, protease ini merupakan protease metaloserin netral berdasarkan uji-uji biokimianya yang dihambat sepenuhnya oleh PMSF (Phenylmethanesulfonyl fluoride) and EDTA (Ethylenediaminetetraacetic acid). Enzim menunjukkan aktifitas optimum pada pH 8 dan aktif lebih baik pada pH bufer bersifat asam (sampai pH 4). . Enzim juga menunjukkan kestabilan terhadap pelarut organik. Nilai K M danV max enzim murni masing-masing adalah 27.2 mg/ml atau 0.362 -0.272 M dengan substrat HammersteinCasein (MM 75-100 kDa) dan 261.1 µg/menit/ ml. Kata Kunci: protease tahan panas, Brevibacillus thermoruber, sumber air panas

Teripang merupakan salah satu echinodermata yang memiliki kandungan protein tinggi dan sekitar 70% dari proteinnya merupakan kolagen. Tujuan penelitian ini adalah untuk mengetahui aktivitas inhibitor Angiotensin Converting Enzyme (ACE) dan antioksidan dari peptida kolagen teripang Gama (Stichopus variegatus). Ekstraksi kolagen dilakukan menggunakan asam asetat 0,5 M. Peptida kolagen diperoleh melalui hidrolisis kolagen menggunakan enzim pepsin dengan konsentrasi 0,1 U/g kolagen, selama 0; 30; 60; 90; 120; 180; dan 240 menit. Aktivitas inhibitor ACE dan antioksidan peptida kolagen diuji dengan metode spektroskopi. Kolagen yang dihasilkan memiliki rendemen 16,40% dengan berat molekul 130,33 kDa. Aktivitas inhibitor ACE tertinggi dihasilkan dari proses hidrolisis selama 180 menit dengan penghambatan sebesar 82,31%, sedangkan aktivitas antioksidan tertinggi dihasilkan oleh peptida kolagen dari hidrolisis kolagen selama 120 menit dengan nilai IC 50 1,9 mg/ml.

Abstract. Zilda DS, Yulianti Y, Sholihah RF, Subaryono S, Fawzya YN, Irianto HE.2019. A novel Bacillus sp. isolated from rotten seaweed: identification and characterization alginate lyase it is produced. Biodiversitas 20: 1166-1172. Alginate lyase has been known as potential biocatalyst not only for industrial but also medicinal application especially for the production of oligosaccharides which have distinct bioactivities. An alginate lyase, AlgT513, has been isolated from rotten seaweed bacterium strain T513 and characterized. The bacterium showed low similarity (95%) with Bacillus tequilensis strain 10b based on 16S rDNA sequence indicating that Alg07 may be a novel Bacillus species. The bacterium forms a clear zone on solid medium with 0.5% sodium alginate addition. The optimum temperature and pH were 50ºC and 8 respectively. AlgT513 maintained stability at board pHs of 4-9 and temperature of 45ºC. Metal ions Mg2+, Ca2+ and K+ increase the activity of the enzyme while Zn2+, Co2+ and Li+ strongly inhibit it. NaCl inhibits AlgT513 activity where most of the alginate lyases need it to reach maximum activity. AlgT513 is suggested as a serine metalloenzyme due to inhibition of ethylenediaminetetraacetic acid (EDTA) and phenylmethylsulfonyl fluoride (PMSF).