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Optimization of Production Xylanase from Marine Bacterium Bacillus safensis P20 on Sugarcane Baggase by Submerged Fermentation Rahmani, Nanik; Jabbar Robbani, Nadia Ulfa; Herawati Suparto, Irma; Yopi, Yopi
International Journal on Advanced Science, Engineering and Information Technology Vol 4, No 6 (2014)
Publisher : International Journal on Advanced Science, Engineering and Information Technology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1309.998 KB)

Abstract

Endo-1, 4-β-xylanase commonly called xylanase is an industrially important enzyme which degrades of lignocellulosic materials to sugar, alcohol and other useful product. The use of commercially xylan is too expensive for use at industrial scale production. For commercial applications, xylanases should ideally be produced from simple and inexpensive substrates. Indonesia has abundantly agro-residues such as sugar cane bagasse which is attractive to be used as carbon sources for the production of enzyme.  In this study, optimization of fermentation condition extracellular xylanasefrom marine bacterium, Bacillus safensisP20 has been conducted by using sugarcane bagasse as carbon source under sub merged fermentation (SMF). Maximum xylanase production was obtained at sugar cane bagasse concentration 1.5%, pH medium 7, and temperature fermentation 20oC, lactose as a carbone source and urea as a nitrogen source with activity 4.06 U/mL for 96 hours.
SELEKSI DAN IDENTIFIKASI BAKTERI ENDOFIT POTENSIAL PENGHASIL ENZIM PROTEASE DARI TAMAN NASIONAL GUNUNG HALIMUN - (The Selection and Identification of Potential Endophyte Bacteria as Protease Enzyme Producer from Halimun Mount National Park) Melliawati, Ruth; Rohmattusolihat, Rohmattusolihat; Nuryati, Nuryati; Rahmani, Nanik; Yopi, Yopi
Biopropal Industri Vol 7, No 2 (2016)
Publisher : Balai Riset dan Standardisasi Industri Pontianak

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Abstract

Endophytic bacteria have an equal chance to bacteria that live outside the plant tissue as potential bacteria. The selection has done towards 326 bacterial endophyte isolates. This research aimed to find and identify proteolytic potential isolates. The proteolytic selection of endophytic bacteria had done using solid skim milk. The capability of endophytic bacteria to agglomerate milk was tested using liquid skim milk which incubated for 7 days at room temperature. Enzyme production of four selected isolates was made through fermentation in GYS medium. The results showed that 86 isolates have proteolytic potential. Isolate HL.29B.63 had highest protease enzymes activity (65.918 U/mL). Medium optimization was able to increase the enzyme activity into 89.94% (125.04 U/mL). The analysis used 16s rDNA showed that isolate HL.29B.63 was Bacillus amyloliquefacient subs. plantarum strain FZB42.Keywords: endophytic bacteria, fermentation, identification, protease, selection ABSTRAKBakteri endofit mempunyai peluang yang sama dengan bakteri yang hidup diluar jaringan tanaman sebagai bakteri potensial. Seleksi dilakukan terhadap 326 isolat bakteri endofit. Tujuan penelitian ini adalah mencari isolat yang berpotensi proteolitik dan mengidentifikasinya. Seleksi proteolitik terhadap bakteri endofitik menggunakan skim milk padat. Uji kemampuan bakteri endofitik dalam menggumpalkan susu menggunakan medium skim milk cair yang diinkubasi selama 7 hari pada suhu ruang. Produksi enzim terhadap empat isolat terseleksi dilakukan melalui fermentasi dalam medium GYS. Hasilnya menunjukkan bahwa 86 isolat mempunyai potensi proteolitik. Isolat HL.29B.63 mempunyai aktif enzim protease tertinggi (65,918 U/mL). Optimasi medium dapat meningkatkan aktivitas enzim sebesar 89,94% (125,04 U/mL). Analisis menggunakan 16s rDNA menunjukkan bahwa isolat HL.29B.63 adalah Bacillus amyloliquefaciens subs. plantarum strain FZB42.Kata kunci: bakteri endofit, fermentasi, identifikasi, protease, seleksi
Isolats Bakteri Indigenous Penghasil Milk-Clotting Protease untuk Fermentasi Keju Rahmani, Nanik; Sari, Yana Nurita; Palupi, Nurheni Sri; Yopi, Yopi
JURNAL BIOLOGI INDONESIA Vol 9, No 2 (2013): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (316.729 KB) | DOI: 10.14203/jbi.v9i2.166

Abstract

The aims of this research is to isolation of bacteria that potential to produce of milk clotting protease enzymes fromfermented food that will be used as a substitute for rennet in cheese making. There are five food fermentations suchas tauco, tempeh, red oncom, sticky tape, and pickled mustard greens that are used as a source for isolation of bacteriathat could produce milk clotting protease. The results obtained four isolates proteolytic bacteria from two fermentedfood samples, three isolates bacteria from tauco (TCN 1, TCN 2, TCN 3) and one isolate from pickledmustard greens (DSN 1). Based on 16S rDNA, these isolates were identified as Bacillus sp. Bacterial isolate TCN 1has a milk clotting activity of 29.17 U/mL, whereas bacteria isolates of TCN 2, TCN 3 and DSN 1 have activities of70 U/mL achieved at the 24 hours incubation, respectively. The proteolytic activities of bacteria isolates TCN 1,TCN 2, TCN 3 and DSN 1 at the 24 hours fermentation process were 0.0117 U/mL, 0.0021 U/mL, 0.0150 U/mL,and 0.200 U/mL, respectively. The ratio of milk clotting protease activity and the proteolytic activity for bacteriaisolates TCN 1, TCN 2, TCN 3 and DSN respectively were 5402, 175000, 7292, and 3333. This showed that theenzyme from bacterial isolates TCN 2 can be used as an alternative to rennin in cheese making.Keywords: milk clotting protease, cheese, calf rennet, fermentation food
Pemurnian Parsial dan Karakterisasi Amilase dari Bakteri Laut Arthrobacter arilaitensis LBF-003 Rahmasari, Dianti; Wijanarka, Wijanarka; Pujiyanto, Sri; Rahmani, Nanik; Yopi, Yopi
JURNAL BIOLOGI INDONESIA Vol 12, No 1 (2016): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v12i1.2323

Abstract

Starch is an abundant carbon source in nature, and ?-amylase (1, 4-?-D-glucanohydrolase; EC 3.2.1.1), which hydrolyzes ?-1, 4-glucosidic linkage in starch-related molecules. Microbe ?-amylase production is a hydrolytic enzyme and one ofinterest in its microbial production has increased dramatically due to its wide spread use in food, textile, baking anddetergent industries in recent years. Here we report ?-amylase from marine bacterium which was purified andcharacterized, as well as analyzed its hydrolysis product on starch. The enzyme of Arthrobacter arilaitensis partiallypurified by acetone precipitation with 90% and ion exchange chromatography produced specific activity 0.25 U/mg and0.38 U/mg, and it’s purity rate increased until 1.14 fold compared with former crude extract. Purifed extracelluler amilasehad an optimum activity at temperature 50°C and pH 9.0. An apparent molecular mass was between 50-75 kDa, asestimated by zimogram electrophoresis. Hydrolysis products of this enzyme on starch were maltose, maltotriose andmaltoheptaose.Keywords: alfa amylase, marine bacterium, Arthrobacter arilaitensis, purification, charaterization
Production Of Malto-Oligosaccharides From Cassava Cultivar Kuning Rahmani, Nanik; Andriani, Ade; Yopi, nFN; Hartati, Sri
Jurnal Penelitian Pascapanen Pertanian Vol 12, No 3 (2015): Jurnal Penelitian Pascapanen Pertanian
Publisher : Balai Besar Penelitian dan Pengembangan Pascapanen Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jpasca.v12n3.2015.147-155

Abstract

Characteristic the physic-chemical of Indonesia cassava starch from four cultivated varieties has been conducted for maltooligosaccharide production. Result of proximate analysis of the extracted starch indicated that the extracted starch was quite pure. The purity of the extracted starch was visually confirmed by microscopic analysis by using SEM micrographs at 2500X magnifications show that the integrity of the granules starch as intact. Based on the amylopectin and amylase content showed that one of cultivated variety of cassava, cultivated variety Kuning contain the amylopectin higher than amylase was compared with the other cultivated variety. The next focus research was analysis potential of starch from cultivated variety Kuning for maltooligosaccharide production by enzymatic hydrolysis by ?-amylase from marine bacterium Brevibacterium sp. The optimum hydrolysis condition for cultivated variety Kuning was obtained substrate concentration 4.5% (b/v), comparison of substrate: enzyme 1:2, temperature reaction 30oC with reducing sugars concentration of 13.359 ppm. The hydrolysis products of cassava starch cultivated variety Kuning were maltooligosaccharides mixture, yielding maltose, maltotriose, maltotetraose, maltopentaose. This result showed that cassava starch of cultivated varieties Kuning potential for maltooligosaccharides production. PRODUKSI MALTOOLIGOSAKARIDA DARI UBI KAYU VARIETAS KUNINGKarakteristik fisiko kimia karbohidrat dari empat varietas kultivar asal Indonesia dilakukan untuk melihat potensinya sebagai bahan baku untuk produksi maltooligosakarida. Analisa proksimat karbohidrat hasil ekstraksi dari keempat varietas kultivar ubi kayu mengindikasikan bahwa karbohidrat yang dihasilkan cukup murni. Kemurnian dari karbohidrat tersebut terlihat setelah dikonfirmasi dengan analisa mikrokospis dengan menggunakan mikroskop electron SEM dengan pembesaran 2500X yang menunjukkan bahwa granulanya utuh. Berdasarkan kadar amilopektin dan amilosa menunjukkan bahwa salah satu varietas kultivar ubi kayu yaitu varietas Kuning mengandung amilopektin lebih tinggi dibandingkan kadar amilosanya jika dibandingkan dengan tiga varietas kultivar lainnya. Fokus penelitian selanjutnya adalah analisa potensi karbohidrat varietas kultivar Kuning tersebut untuk produksi maltooligosakarida dengan hidrolisis enzimatis oleh ?-amylase dari Brevibacterium sp. Kondisi optimum hidrolisis dari varietas kultivar Kuning diperoleh pada konsentrasi substrat 4.5% (b/v), perbandingan substrat dan enzim 1:2, suhu reaksi 30oC dengan kadar gula reduksi yang diperoleh 13.359 ppm. Produk hidrolisis dari ubi kayu varietas kultivar Kuning adalah berupa campuran maltooligosakarida yang terdiri atas maltose, maltotriose, maltotetraose, maltopentaose. Hasil ini menunjukkan bahwa karbohidrat ubi kayu dari varietas kultivar Kuning mempunyai potensi untuk digunakan sebagai bahan baku untuk produksi maltooligosakarida.
Pemurnian Parsial dan Karakterisasi Enzim Xilanase dari Bakteri Laut Bacillus safencis strain LBF P20 Asal Pulau Pari Jakarta Fitria, Fitria; Rahmani, Nanik; Pujiyanto, Sri; Raharjo, Budi; Yopi, Yopi
Agritech Vol 37, No 1 (2017)
Publisher : Faculty of Agricultural Technology, Universitas Gadjah Mada, Yogyakarta, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (638.397 KB) | DOI: 10.22146/agritech.17004

Abstract

Enzyme xylanase (EC 3.2.1.8) is widely used in various industrial  fields for the hydrolysis of xylan (hemicellulose) into xylooligosaccharide and xylose. The aims of this study were to  conduct partial purification and characterization of xylanase from marine Bacillus safencis strain LBF P20 and to obtain the  xylooligosaccharide types from xylan hydrolysis by this enzyme.  Based on this research, the optimum time for enzyme production  occurred at 96 hours with the enzyme activity of 6.275 U/mL and  enzyme specific activity of 5.093 U/mg. The specific activities were  obtained from precipitation by amicon® ultra-15 centrifugal filter devices, gel filtration chromatography and anion exchange chromatography that were increased by 15.07, 34.7, and 96.0  U/mg. The results showed that the highest activity at pH 7, temperature of 60 °C, and stable at 4 °C. Type of  xylooligosaccharide produced by this study were xylohexoses, xylotriose, and xylobiose. SDS-PAGE analysis and zimogram  showed that the molecular weight of xylanase protein were about  25 kDa. ABSTRAKEnzim xilanase (EC 3.2.1.8) digunakan dalam hidrolisis xilan  (hemiselulosa) menjadi xilooligosakarida dan xilosa. Penelitian  ini bertujuan untuk melakukan purifikasi parsial dan karakterisasi xilanase dari bakteri laut Bacillus safencis strain LBF P20 serta uji  hidrolisis untuk mengetahui jenis xilooligosakarida yang  dihasilkan oleh enzim tersebut. Berdasarkan hasil penelitian, waktu optimum untuk produksi enzim terjadi pada jam ke 96  dengan aktivitas enzim sebesar 6,275 U/mL dan aktivitas spesifik enzim sebesar 5,093 (U/mg). Aktivitas spesifik enzim hasil  pemekatan dengan amicon® ultra-15 centrifugal filter devices,  kromatografi filtrasi gel dan kromatografi penukar anion  mengalami peningkatan berturut-turut sebesar 15,1; 34,7 dan96,0 U/mg. Hasil karakterisasi menunjukkan aktivitas  tertinggi pada pH 7, suhu 60 °C dan stabil pada suhu 4 °C. Analisis SDS-PAGE dan zimogram menunjukkan berat molekul protein xilanase berkisar 25 kDa. Jenis gula reduksi yang  dihasilkan yaitu xiloheksosa, xilotriosa, dan xilobiosa.
The Optimal Conditions of Xylanase Production Using Empty Fruit Bunch Raw Biomass by Marine Isolate LBF-001 Wijaya, Hans; Thontowi, Ahmad; Rahmani, Nanik; Yopi, Yopi
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.v11i3.211

Abstract

Several xylanases have already been studied. However, only a few xylanases derived from marine microorganisms has been reported. Marine bacterium LBF-001 was isolated from Pari Island Kepulauan Seribu, Indonesia. The purpose of this study is to identify isolate LBF-001 using 16S rDNA gene and to optimise the medium conditions for the xylanase production i.e. concentration of biomass, nitrogen source, pH and temperature. Based on 16S rDNA gene analyses, LBF-001 isolate has 99% similarity with Bacillus pumilus HNS70 (KF933667). Fermentation process to produce xylanases was conducted using several agricultural residues under solid-state fermentation (SSF). The optimum condition for xylanase production by B. pumilus LBF-001 was using a medium containing  2.5% empty fruit bunch and 0.6% lactose broth, at pH 6.5, temperature 30 oC, under submerged fermentation with shaking at 150 rpm for 48 h fermentation. The optimised condition resulted higher xylanase activity, i.e. 10.85 U/mL.
Cloning of Partial β-Mannanase Gene from Indonesia Marine Bacteria Bacillus Subtilis LBF-005 yopi, Yopi; Rahmani, Nanik; Amaniyah, Maghfirotul; Djohan, Apridah Cameliawati
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.v12i1.252

Abstract

 The strain LBF-005 from marine bacteria have already isolated and screened for mannanase degrading enzyme in submerged fermentation process. This strain was further identified by using 16S rRNA showed that bacterium is belong to Bacillus subtilis that could produce mannanase with activity around 9.5 U/mL. The optimum pH and temperature for the activity of crude enzyme for mannanase was 6.0 and 50 oC. Cloning of mannanase gene from B. subtilis was conducted using six primers set designed based on the homology analysis conserve region several mannanase from bacteria (Bacillus sp.) glycosyl hydrolase (GH) family 26. Optimization of PCR conditions was performed by gradient PCR to obtained PCR product of b-mannanase gene. The PCR product was obtained by third primer combination and was estimated to be around 972-bp. Analysis of the nucleotide sequence showed that sequences has similarity with mananase gene from other Bacillus sp., such as the B. subtilis strain WLY-12, B. subtilis strain WL-8, B. subtilis strain CICC 10260, B.subtilis strain CD-25, B.subtilis strain G1, and Bacillus sp. SWU60 were about 98%, 98%, 98%, 98%, 97% and 95%, respectively. The next research will to obtain a whole gene encoding β-mannanase by using in vitro cloning method, characterization of recombinant mannanase and application this enzyme for mannooligosaccharides production.
Polyaromatic Hydrocarbon Degradation and Dioxygenase Gene Detection from Alteromonas alvinellae Bt05 Thontowi, Ahmad; Rahmani, Nanik; Yopi, Yopi
ANNALES BOGORIENSES Vol 17, No 1 (2013): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.1234/75

Abstract

Bt05 is marine bacterium which was isolated from the Jakarta Bay, Indonesia. The aim of this study was to characterize PAHs-degrading property, molecular identification by partial analysis of 16S rRNA gene and to partially analyze dioxygenase gene of Bt05 isolate. Our further study on this isolate revealed that it could degrade three PAHs (phenanthrene, dibenzothiophene, fluorene) between 60%–90% within 11 days at 100 ppm level. This finding indicated the potential of the isolate for bioremediation of PAHs. The isolate was identified as Alteromonas alvinellae by phylogenetic analysis of 16S rRNA gene sequence. Sequence analysis of the PCR product of PAH dioxygenase genes amplified using two primer set (iiDA and ppAH) of the isolate were identified 97% as naphthalene dioxygenase gene (phaAc) and 58% as 1,2-dioxygenase.
OPTIMIZATION OF CELLULASE PRODUCTION FROM MARINE BACTERIUM BACILLUS CEREUS C9 BY SUBMERGED FERMENTATION -, Yopi -; Rahmani, Nanik
Teknologi Indonesia Vol 39, No 1 (2016)
Publisher : LIPI Press

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (0.001 KB) | DOI: 10.14203/jti.v39i1.238

Abstract

Cellulase is one of industrial important enzymes in conversion of lignocellulosic biomass into valuable products such as bioethanol produced by fermentation. Successful bioconversion of cellulosic biomass mainly depends on the nature of cellulose sources of cellulolytic enzyme and optimal conditions for production of enzymes. An extracellular cellulase production by marine bacterium Bacillus cereus C9 was optimized by using submerged fermentation (SMF) on commercial substrate (CMC). The fermentation condition such as substrate concentration, pH medium, temperature fermentation, and carbon source were optimized. The optimum condition found for cellulase production were substrate concentration 2.0% (b/v), pH medium 5, temperature fermentation 30C, and glucose as a carbon source with activity 4.42 U ml-1on 96 hours of fermentation.