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All Journal Jurnal Natur Indonesia Biota: Jurnal Ilmiah Ilmu-Ilmu Hayati BioWallacea Journal of Biological Research Indonesian Journal of Biotechnology
Sebastian Margino
Mikrobiology Laboratory, Faculty of Agriculture, Universitas Gadjah Mada. Yogyakarta, Indonesia.
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Immunology & microbiology Materials Science & Nanotechnology Veterinary

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A Study on Production of Poly-β-Hydroxybutyrate Bioplastic from Sago Starch by Indigenous Amylolytic Bacteria Yanti, Nur Arfa; Sembiring, Langkah; Margino, Sebastian; Muhiddin, Nurhayani H.
Indonesian Journal of Biotechnology Vol 18, No 2 (2013)
Publisher : Universitas Gadjah Mada

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Abstract

Bacillus sp. PSA10 and Bacillus sp. PPK5 were two indigenous strain amylolytic bacteria from SoutheastSulawesi that have ability to produce bioplastic poly-β-hydroxybutyrate (PHB) from sago starch. The study wasattempted to determine the mechanism of PHB production by bacteria amylolytic was grown on sago starchcontainingmedia. Two amylolytic bacteria i.e. Bacillus sp. PSA10 and Bacillus sp. PPK5 was grown for 168 hin a mineral salts medium with sago starch as carbon source. Growth of amylolytic bacteria was monitoredby cell dry weight. Extraction of PHB was done by N-hexane acetone-diethyl ether method and PHB contentwas quantifi ed with UV spectrophotometer at 235 nm. Glucose level was determined by using kit of glucoseGOD 10” and was quantifi ed with spectrophotometer at 500 nm. Sago starch concentration was determinedby phenol method using specthrophotometer at 490 nm. The result of the study showed that Bacillus sp.PSA10 was produced PHB up to 66,81 % (g PHB/g cell dry weight) at 48 h and Bacillus sp. PPK5 up to 24,83% (g PHB/g cell dry weight) at 84 h. Bacillus sp. PSA10 has ability to converse sago starch to be PHB directlywithout glucose accumulation in the media, whereas Bacillus sp. PPK5 have to accumulate glucose as productof sago starch hydrolysis to produce of PHB. PHB synthesis by Bacillus sp. PHB production on sago starchof the Bacillus sp. PSA10 was found to be growth-associated whereas Bacillus sp. PPK5 was found to be nongrowth-associated. Therefore, two indigenous amylolytic bacteria were having of difference in biosynthesismechanism of PHB in sago starch medium and their characteristics of PHB synthesis should be consideredin developing cultivation methods for the effi cient production of PHB.Keywords : Production, PHB, Amylolytic bacteria, Sago starch.
Production of Poly--hydroxybutyrate (PHB) from Sago Starch by The Native Isolate Bacillus megaterium PSA10 Yanti, Nur Arfa; Sembiring, Langkah; Margino, Sebastian
Indonesian Journal of Biotechnology Vol 14, No 1 (2009)
Publisher : Universitas Gadjah Mada

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Abstract

A new bacterial strain that produces amylase and poly-a-hidroxybutyrate (PHB) using sago starch as carbon source was characterized and identified to be member of the Bacillus megaterium group based on phenotypic characteristics  and 16S rDNA gene sequences. Amylase activity was determined spectrophotometrically on the basis of substrate concentration reduction. PHB production was quantified with UV spectrophotometer. The polymer produced by B. megaterium PSA10 was identified by  Fourier Transform Infrared spectroscopy (FTIR). The result of the study showed that the amylase specific activity B. megaterium PSA10 was 593,61 DUN/mg protein and PHB production from sago starch was 52,28 % of cell dry weight (CDW). FTIR analysis of the polymer indicated that the strain B.megaterium PSA10 was a potent PHB producer.
Purifi cation and Characterization of Protease From Bacillus sp. TBRSN- 1 Margino, Sebastian; ., Ngadiman
Indonesian Journal of Biotechnology Vol 18, No 2 (2013)
Publisher : Universitas Gadjah Mada

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Abstract

Potato Cyst Nematode (PCN), Globodera rostochiensis, is one of the important potato’s pests and causedeconomic looses up to 70% in the several centrals of potato plantations in Indonesia. PCN’s shell componentof egg shell containing chitin (inner layer) and viteline/ protein (outer layer). The purpose of this researchwas to purify of protease Bacillus sp. TBRSN-1, isolate from tomato’s rhizosfer in Yogyakarta province. Thepurifi ed protease could be used for cutting the life cycle of PCN. Results showed that Bacillus sp. TBRSN-1could produce extracellular protease and purifi cation using DEAE-cellulose ion-exchange chromatographyand Sephacryl S-300 gel fi ltration chromatography resulted in specifi c activity 4.31 fold and 1.68% recovery.Analysing using SDS-PAGE 12.5% and molecular weight 48.1 kDa. Km and Vmax values of the protease forcasein substrate were 7.83 mg/ml and 4.03 μg/h, respectively. The optimum activity at the temperature30oC and pH 7.0.Keywords : protease, purifi cation, indigenous Bacillus sp. TBRSN-1
Isolation and Purifi cation of Chitinase Bacillus sp. D2 Isolated from Potato Rhizosfer Margino, Sebastian; Behar, Chatarina; Asmara, Widya
Indonesian Journal of Biotechnology Vol 17, No 1 (2012)
Publisher : Universitas Gadjah Mada

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Abstract

Potato Cyst Nematodes (Globodera rostochiensis) is one of the important potato’s pests and caused economic looses up to 70% in the several centrals of potato plantations in Indonesia. Potato Cyst Nematodes (PCN) shell component of egg shell containing chitin (inner layer) and vitelline/protein (outer layer), so the purpose of research was to fi nd out of chitin degrading bacteria for controlling of egg’s PCN by cutting of their life cycle. The results showed that Bacillus sp. D2 isolated from potato rhizosphere could produce extra cellular chitinase in the medium containing of 0.20% colloidal chitin and fermented for 72 hours. Result of chitinase purifi cation using ammonium sulphate precipitation and DEAE-Cellulose ion-exchange chromatography showed a specifi c activity 2691,052 U/mg and analyzing using SDS-PAGE 12.5% resulted in molecular weight 30 kDa. The apparent Km and Vmax of chitinase towards colloidal chitin were 2 mg/ml and 2.2 μg/h, respectively.  
Purification and Characterization of Streptomyces sp. IK Chitinase Margino, Sebastian; Nugroho, Agustinus Joko; Asmara, Widya
Indonesian Journal of Biotechnology Vol 15, No 1 (2010)
Publisher : Universitas Gadjah Mada

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Abstract

Streptomyces sp. IK isolated from compost inoculants, could produce extra cellular chitinase in a medium containing 0.2% (w/v) colloidal chitin, fermented for 96 hours at 30oC. The enzyme was purified by a combination of ammonium sulphate precipitation and DEAE-Cellulose anion-exchange chromatography. On SDS-polyacrylamide gel electrophoresis analysis, the purified enzyme showed a mass of 71 kDa. Chitinase was optimally active at pH of 6.7 and at 37oC. Km value and Vmax of the protein for colloidal chitin were 2.92 mg/ml and 4.26 ìg/h, respectively.Key words : chitinase, Streptomyces, purification, characterization
Poly-β-Hydroxybutyrate (PHB) Production By Amylolytic Micrococcus sp. PG1 Isolated From Soil Polluted Arrowroot Starch Waste Margino, Sebastian; Martani, Erni; Prameswara, Andriessa
Indonesian Journal of Biotechnology Vol 19, No 1 (2014)
Publisher : Universitas Gadjah Mada

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Abstract

Poly-β-hydroxybutyrate (PHB) production from amylolytic Micrococcus sp. PG1. Poly-β-hydroxybutyrate(PHB) is an organic polymer, which synthesized by many bacteria and serves as internal energy. PHB ispotential as future bioplastic but its price is very expensive due to glucose usage in PHB industry. Thedevelopment of PHB production using starch as an alternative carbon source has been conducted to reducethe dependence of glucose in PHB production. In this study, amylolytic bacteria from arrowroot processingsite were screened quantitavely based on amylase specifi c activity and PHB producing ability. The result of thestudy showed that among of 24 amylolytic isolates, 12 isolates of them were able to accumulate PHB rangedfrom 0,68-11,65% (g PHB/g cdw). The highest PHB production from substrate arrowroot starch was PG1 andafter optimization resulted in increasing of PHB production up to 16,8% (g PHB/g cdw) 40 hours incubationtime. Based on morphological, biochemical and physiological characters, the PG1 isolate was identifi ed asMicrococcus sp. PG1. Result of the FTIR analysis of produced polymer by Micrococcus sp. PG1 was indicatedas poly-β- hydroxybutyrate (PHB)
Production of Poly-α-hydroxybutyrate (PHB) from Sago Starch by The Native Isolate Bacillus megaterium PSA10 Yanti, Nur Arfa; Sembiring, Langkah; Margino, Sebastian
Indonesian Journal of Biotechnology Vol 11, No 1 (2006)
Publisher : Universitas Gadjah Mada

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Abstract

A new bacterial strain that produces amylase and poly-α-hidroxybutyrate (PHB) using sago starch as carbon source was characterized and identified to be member of the Bacillus megaterium group based on phenotypic characteristics and 16S rDNA gene sequences. Amylase activity was determined spectrophotometrically on the basis of substrate concentration reduction. PHB production was quantified with UV spectrophotometer. The polymer produced by B. megaterium PSA10 was identified by Fourier Transform Infrared spectroscopy (FTIR). The result of the study showed that the amylase specific activity B. megaterium PSA10 was 593,61 DUN/mg protein and PHB production from sago starch was 52,28 % of cell dry weight (CDW). FTIR analysis of the polymer indicated that the strain B.megaterium PSA10 was a potent PHB producer.
Superoxide Dismutase of Micrococcus sp. S2 and Its Involve in Paraquat Detoxification Margino, Sebastian; Martani, Erni; Magdalena, Medhina
Indonesian Journal of Biotechnology Vol 12, No 1 (2007)
Publisher : Universitas Gadjah Mada

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Abstract

As an active ingredient of herbicide, paraquat will induce formation of superoxide radicals. The previousresearch succeeded in isolating paraquat degrading bacteria from peat soil, Micrococcus sp. S2, that tolerant to highconcentration of paraquat. An anti-oxidative enzyme, namely superoxide dismutase (SOD, EC.1.15.1.1), wasbelieved to be responsible for the paraquat tolerance. This research was conducted to study the characteristic of theSOD synthesize by Micrococcus sp. S2 and its ability on neutralize superoxide which arise from paraquat reoxidation.To observe the effect of paraquat on Micrococcus sp. S2, the bacteria was grown in 10% Luria Bertani brothmedium amended with several concentrations of paraquat, from 0 (control) up to 100 mg/ml. Within incubationtime of 72 hours, bacterial growth, activity of superoxide dismutase and paraquat residue were analyzed. Theisozymes of superoxide dismutase were distinguished using two kinds of specific inhibitor, namely HO and KCN. 2 2The results showed that paraquat significantly inhibit the growth of Micrococcus sp. S2. The higher paraquatcocentration in the medium caused the higher growth inhibition. However, the bacteria is still survive in the mediumcontaining toxic herbicide, and this ability was suggested related to superoxide dismutase activity in removing thesuperoxide radicals. Analysis using gel electrophoresis indicated that at least three types of SOD isozyme weresynthesized by Micrococcus sp. S2; they were Ferri-SOD (Fe-SOD), Mangani-SOD (Mn-SOD), and the last one wassuspected to be the Cupro Zinc-SOD (CuZn-SOD). The Mangani-SOD was suspected to play an important roles ondetoxifying superoxide which arise from paraquat oxidation.Keywords : Micrococcus sp.S2, paraquat, superoxide dismutase, isozymes
PENGHASILAN BIOPLASTIK OLEH ISOLAT INDIGENUS Bacillus sp. AMILOLITIK DENGAN SUBSTRAT PATI SUWEG (Amorphophallus campanulatus) Margino, Sebastian; Sari, Rarat Mulat; Martani, Erni
Jurnal BioWallacea Vol 2, No 1 (2015): Bodiversitas
Publisher : Jurnal BioWallacea

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Abstract

bioplastic that was synthesized by bacteria. Glucose as the main carbon source for PHBproduction but its cost was expensive, so we need to substitute with another substrate,including starch.The ability of amylolytic bacteria hydrolyzed starch into simple sugars andconverted to PHB can minimize the costs of PHB production. This research aim was to findout amylolitic bacterial isolates and produce PHB using the selected isolate. A total of 52bacteria were isolated using starch mediumfrom soil sample around Amorphophalluscampanulatus plant. Twenty eight among of them had amylase activity, and eleven among ofthem had amylase activity value equal or more than 2 point.The next selection was donebased on the specific amylase activity and found that five of those isolates had specificamylase activity more than 10 DUN/mg.Final selection was done based on PHB productionand found that isolate E5 had the high amylase activity and PHB production. PHB productionused isolate E5 in optimum conditions (inoculum concentration 10 %, elephant foot yamstarch 3 %, medium pH 7, temperature 30 °C, agitation 125 rpm, and 48 hour incubationtimeshowed that percentage of PHB increased from 9,5% to 14,5%. Characterization andidentification showed that isolate E5 closed to the genus Bacillus sp.Keywords : Amylolytic Bacillus sp., poly-β-hidroxybutyrate, elephant foot yam starch.
Kemampuan Kitinase Streptomyces RKt5 sebagai Antijamur terhadap Patogen Fusarium oxysporum Yurnaliza, Yurnaliza; Margino, Sebastian; Sembiring, Langkah
Jurnal Natur Indonesia Vol 14, No 1 (2011)
Publisher : Lembaga Penelitian dan Pengabdian kepada Masyarakat Universitas Riau

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (272.537 KB) | DOI: 10.31258/jnat.14.1.42-46

Abstract

The purpose of the reasearch is to determine of antifungal activity from chitinase from Streptomyces RKt5 to inhibite growth of Fusariumoxysporum. The chitinase of Streptomyces RKt5 produced in liquid chitin medium with optimum conditions (inoculum concentration, pHand incubation time) and then partially purified with ammonium sulphate. The enzyme products were tested the antifungal activity againstF.oxysporum. The results showed that mycelial growth of F.oxysporum can be inhibited by Streptomyces RKt 5 in dual culture test. Thepartial purified chitinase enzyme couldn’t inhibit the fungal growth. But if the mycellium fragmented, the enzyme could degrade the fungalcell wall in incubation time. The frequency of fungal cell wall lysis and levels of N-acetylglucosamine released that have been increasingalong with the length of incubation time.
Co-Authors Agustinus Joko Nugroho, Agustinus Joko ANDRIESSA PRAMESWARA Bostang Radjagukguk Chatarina Behar, Chatarina Elisa Nurnawati Erni Martani Kurniawan Wibowo, Kurniawan LANGKAH SEMBIRING Medhina Magdalena, Medhina Ngadiman ., Ngadiman Nur Arfa Yanti, Nur Arfa Nurhayani H. Muhiddin, Nurhayani H. Sari, Rarat Mulat Sarto Widya Asmara Yurnaliza, Yurnaliza