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All Journal Squalen Bulletin of Marine and Fisheries Postharvest and Biotechnology INDONESIAN JOURNAL OF MEDICAL LABORATORY SCIENCE AND TECHNOLOGY
Dewi Seswita Zilda
Balai Besar Penelitian dan Pengembangan Pengolahan Produk dan Bioteknologi Kelautan dan Perikanan Jl. KS Tubun Petamburan VI, Jakarta, Indonesia. 11420
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Environmental Science Health Professions Immunology & microbiology Social Sciences

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Metagenomics-Based Cloning of Amilase-Encoding Genes from the Uncultured Symbiotic Bacteria of a Marine Sponge Theonella swinhoei from Kapoposang Island, South Sulawesi Priyono, Franciscus Edi; Zilda, Dewi Seswita; Kusnadi, Yudi; Hadi, Tri A; Nurrachmi, Irvina; Uria, Agustinus Robert
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 12, No 1 (2017): May 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.v12i1.272

Abstract

Marine sponges have recently been recognized as the source ofenzymes, including members of hydrolases. Hydrolytic enzymes are extracellularly produced by sponge-associated bacteria to mediate the metabolism of complex organic matters, thereby assisting the sponge hosts in nutrition and metabolic processes. Among hydrolytic enzymes, amilaseshas attracted increasing attention due to their potential industrial applications. This research work was aimed atutilizing functional metagenomicsapproach for the discovery of amilases derived from the uncultured symbiotic bacteriaof the Indonesian marine sponge Theonella swinhoei. Weinitially constructed a small-insert metagenomiclibrary in Escherichia coliby cloning of metagenome in the size range of5-20 kb prepared from the sponge’s microbiome. Further functional screening of the resulting metagenomic library led to the isolation of two recombinant E. coli clones potentially harboring amilase genes, as indicated by the presence of clearing zones surroinding the selective medium containing 1% amilum. 
Identification of Protease-Producing Bacteria Isolated from Banyuwedang, Bali, and Characterization of its Protease Zilda, Dewi Seswita; Fawzya, Yusro Nuri; Uria, Agustinus Robert
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 13, No 3 (2018): December 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.v13i3.367

Abstract

Proteases or peptidases is known as a largest group of hydrolytic enzymes and have been applied in various industries such as food, pharmacy, leather, detergent and waste treatment. Although they are also produced by plants and animals, microbes remain the main source of proteases in the world market which mostly derived from Bacillus sp. Aims of this research were to identify isolate BII-1 and study its protease. Analysis of 16Sr RNA sequencing showed the identity of BII-1 as Bacillus subtilis (99% similarity with the same species in GenBank). It was found that protease from BII-1 exhibited optimal temperature and pH of 50 oC and 8-9, respectively. It was activated by Li2+, Na2+, Mg2+ and K+. The degenerated primer for protease gene was designed, and a partial protease gene was amplified from BII-1. The sequencing result showed that this amplified gene shared 100 and 99% similarity with those from Geobacillus thermophiles and Bacillus subtilis in the GenBank, respectively.Keywords: protease, bacteria, Bacillus subtilis, Geobacillus thermophylus
In Vitro Anticoagulant Activity of Crude Protease of Bacillus tequilensis HSFI-5 Ethica, Stalis Norma; Raharjo, Tri Joko; Zilda, Dewi Seswita; Hidayati, Nur
JURNAL INDONESIA DARI ILMU LABORATORIUM MEDIS DAN TEKNOLOGI Vol 5 No 2 (2023): Combatting Bacterial and Fungal Infections: The Critical Role of Advanced Researc
Publisher : Universitas Nahdlatul Ulama Surabaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33086/ijmlst.v5i2.3791

Abstract

obtained from the fermented intestine of Holothuria scabra (sand sea cucumber). Strain HSFI-5 had been reported to be able to produce proteases, which had shown several characteristics of an antithrombotic agent, i.e., fibrinolytic and clot-lysis activities. However, its anticoagulation activity test is yest to be done. This study aimed to determine the anticoagulant activity of the crude protease HSFI-5 in vitro. The study design was a completely randomized design with a sample size of 90 calculated using the Federer formula. The material used was crude protease from B. tequilensis in skim milk broth. Prothrombin time (PT), activated partial thromboplastin time (aPTT), and plasma recalcification time (PRT) were carried out to test the anticoagulant activity. Citrated platelet poor plasma samples were divided into positive control, normal control, direct examination with crude enzyme in volumes of 50 and 100 µL and pre-incubation at 37ºC for 5, 10, and 15 min with crude enzyme volumes of 50 and 100 µL. The data normality was tested with the Kolmogorov-Smirnov test and the different tests were analyzed by one-way ANOVA with the Post hoc LSD test. The results of one-way ANOVA both on PT, aPTT, and PRT examinations showed that there was a significant difference between the treatment groups (p<0.05). The longest results of PT, aPTT, and PRT are positive controls, and the shortest results are normal controls for PT, and 15’ 50 group for aPTT and PRT. It is clear that crude protease B. tequilensis HSFI-5 exhibits anticoagulant as well as thrombolytic action, raising the possibility that it could function as an antithrombotic drug.
Antimicrobial ointment based on Bacillus subtilis subsp. subtilis HSFI-9 isolated from Sea Cucumber of Kodek Bay Lombok Indonesia Rakhmawatie, Maya Dian; Diatri, Devita; Samiroh, Samiroh; Abdillah, Rifqi; Arfiyanti, Mega Pandu; Ethica, Stalis Norma; Zilda, Dewi Seswita
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 20, No 1 (2025): May 2025
Publisher : :Agency for Marine and Fisheries Research and Human Resources, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.925

Abstract

The ethyl acetate extract of Bacillus subtilis subsp. subtilis HSFI-9, a bacteria isolated from intestinal fermentation of sea cucumbers (Holothuria scabra) is known to have antimicrobial properties. This research aimed to determine the activity of HSFI-9 extract ointment against Staphylococcus aureus and Candida albicans. Extract ointment is a topical preparation for skin infections made from a mixture of Vaseline Alba and Adeps lanae bases. The extract ointment was prepared into four concentrations of 0.003%, 0.03%, 0.3%, and 1% w/v using HSFI-9 as active ingredient.  Antimicrobial assay was carried out in vitro using the disc diffusion method. The extract ointment was evaluated based on organoleptic, homogeneity, spreadability, adhesion characteristics, and pH tests. The optimal concentration of the extract ointment to inhibit the growth of S. aureus  0.3% (inhibition zone of 11.67±1.26 mm) and against C. albicans is 0.03% (inhibition zone of 10.16±1.50 mm). The activity of the extract ointment was categorized as strong although not as strong as the antibiotic control ointment Mupirocin 2% or Ketoconazole 2%. The extract ointment organoleptic indicated a characteristic odor of ethyl acetate, was yellowish-white ointment, and had a homogeneous and smooth consistency. The extract ointment also had properties such as good spreadability but poor adhesion and tended to have an acidic pH ( 4.5). The HSFI-9 extract can be declared feasible for the development of topical antimicrobials. The ointment still needs to be optimized, especially regarding improving the adhesion characteristic and pH to be safe for the skin and mucosa.
Co-Authors Abdillah, Rifqi Agustinus Robert Uria Arfiyanti, Mega Pandu Diatri, Devita Irvina Nurachmi Kusnadi, Yudi Maya Dian Rakhmawatie Nur Hidayati Priyono, Franciscus Edi Samiroh Samiroh, Samiroh Stalis Norma Ethica Tri A Hadi Tri Joko Raharjo Yusro Nuri Fawzya