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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

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (612.616 KB)

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.
Identification of SGS 1609 Cellulolytic Bacteria Isolated from Sargassum spec. and Characterization of The Cellulase Produced Yusro Nuri Fawzya; Stenny Putri; Nita Noriko; Gintung Patantis
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 8, No 2 (2013): August 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.v8i2.87

Abstract

Bacterial isolate from seaweed designated as SGS 1609 was previously found to be able to produce cellulase represented by formation of clear zone on solid medium containing carboxymethylcellulose (CMC). This research was conducted to identify the isolate and determine optimum production time as well as characterize the cellulase produced. The isolate was identified using  16s-rRNA gene analysis. Cellulase production was conducted by cultivating the isolate in the liquid medium containing CMC followed by centrifuging to get supernatant as the crude enzyme. The enzyme was then concentrated using ammonium sulfate precipitation and ultra filtration. The concentrated enzyme having higher activity produced from the concentration process was then characterized  to determine its optimum pH and temperature, heat stabilization, metal ions effect and substrate specificity. The result showed that the SGS 1609 isolate was identified as Serratia marcescens with 99%  similarity. The isolate produced cellulase optimally at 4 days incubation. Ultra filtration produced higher enzyme activity compared to NH4-sulfate precipitation. The enzyme concentrated by ultra filtration worked optimally at the  pH of 7, temperature of 50 oC, stable at the temperature of 60 oC for 240 minutes and was increased its activity by Ca2+ and Mg2+ ions. On the other hand, the enzyme was inhibited by Fe3+, Zn2+ and Na+ ions, but was not relatively affected by K+ and EDTA. The use of conventional agar producer waste  treated with 6% NaOH gave highest activity compared to other substrates.
SCREENING OF THERMOSTABLE PROTEASE PRODUCING MICROORGANISMS ISOLATED FROM INDONESIAN HOTSPRING Dewi Seswita Zilda; Eni Harmayani; Jaka Widada; Widya Asmara; Hari Eko Irianto; Gintung Patantis; Yusro Nuri Fawzya
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.v7i3.5

Abstract

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.
BACTERIAL DIVERSITY OF THE DEEP SEA OF SANGIHE TALAUD, SULAWESI Gintung Patantis; Ekowati Chasanah; Dewi Seswita Zilda; Ikhsan B. Waluyo
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 7, No 1 (2012): May 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.v7i1.12

Abstract

Deep sea is an extreme environment characterized by cold temperature, high pressure, lackof  light and nutrients. Microorganisms live in these habitat are unique microorganisms andknown to have tremendous source of potential agents for biotechnology processes. Indonesia asan archipelagic country has a vast deep ocean. This study aims to see the diversity of bacteria inSangihe Talaud Deep Sea, Sulawesi. Analysis of bacterial diversity was carried out by culturedand uncultured method. Terminal Restriction Fragment Length Polymorphism (T-RFLP) techniquewas used for uncultured analysis of the microorganisms biodiversity, while cultured one wasdone by plating the samples of water onto Zobell media. The results showed that, there were 21isolates obtained by cultured method. The identification which based on 16S rDNA by PCR methodshowed the genus of Pseudomonas, Pseudoalteromonas, Alteromonas, Vibrio, Shewanella andUncultured bacterium were identified. However, 14 classes of bacteria were obtained by usingTRFLP method i.e Acetobacteraceae class, Actinobacteria, α-proteobacteria, -proteobacteria, δ-proteobacteria, γ-proteobacteria, Bacili, Bacteroidetes, Chlorobi, Chroococcales, Clostridia,Erysipelotrichi, Synergistia, and Zetaproteobacteria. here were also  unclassified bacteria anduncultured bacterium found in the samples.
Golden Sea Cucumber: Identification and the Antioxidant Activity of Its Collagen Hydrolysates Yusro Nuri Fawzya; Nugrah Analiadi Putra; Arif Budi Witarto; Gintung Patantis
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 15, No 3 (2020): December 2020
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.v15i3.511

Abstract

Golden sea cucumber or locally known as “teripang emas” is one of Indonesia’s most popular sea cucumber and widely processed as functional food or supplement due to its bioactivities. The sea cucumber is often misidentified due to its morphological similarities with other Stichopus spp. This study aimed to identify the golden sea cucumber obtained from West Nusa Tenggara, Indonesia, by a molecular method and study the antioxidant activities of its collagen hydrolysates. The hydrolysates were produced by hydrolyzing acid collagen extract using neutrase for 30, 60, 120, 180, and 240 mins. The products were then analyzed for their degree of hydrolysis, peptide content, molecular weight distribution and radical scavenging activity by the 2,2-Diphenyl-1-picrylhydrazyl (DPPH) method. Results showed that hydrolysis for 180 mins was optimal in producing the highest peptide content, 12.79 ± 0.44 mg/mL, with a degree of hydrolysis (DH) of 55.2 ± 1.50%. However, the highest antioxidant activity (IC50 of 5.25 ± 0.15 mg/mL) was demonstrated after 60 mins hydrolysis with molecular weight (MW) ranged from less than 14.4 kDa to approximately 25 kDa. The hydrolysate might be categorized as a weak to moderate antioxidant. Based on the molecular identification, the golden sea cucumber had 99% similarities with Stichopus horrens and S. monotuberculatus.