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Abstrak :
[Telah dilakukan penelitian deteksi gen alkana monooksigenase (alkB) pada bakteri laut di Perairan Pulau Pari Kepulauan Seribu, Jakarta. Penelitian bertujuan untuk memperoleh isolat bakteri yang membawa gen alkB dari perairan Pulau Pari Kepulauan Seribu, Jakarta. Penelitian dilakukan selama 5 bulan sejak bulan Februari 2015 sampai bulan Mei 2015 dengan metode Polymerase Chain Reaction (PCR) pada 81 isolat yang telah diremajakan. Isolat bakteri diremajakan menggunakan medium marine agar (MA) dengan metode kuadran streak. Hasil deteksi mendapatkan satu isolat yang membawa gen alkB yaitu isolat nomor 71. Hasil amplifikasi isolat 71 menghasilkan pita DNA dengan ukuran 550 pb. Pita DNA dengan panjang 550 pb merupakan gen alkB. Hasil dari sekuensing menunjukkan bahwa Isolat 71 adalah dari spesies Bordetella sp. ;Detection gene alkane monooxygenases (alkB) from marine bacteria in Pari Island Kepulauan Seribu, Jakartahas been researched. The research aims to obtain bacterial isolates that carry the gene alkBin Pari Island Kepulauan Seribu, Jakarta. The study was conducted during the five months from February 2015 to May 2015 with a method of Polymerase Chain Reaction (PCR) from 81 isolates that have been rejuvenated. Bacterial isolates rejuvenated using marine medium agar (MA) with the quadrant streak method. Obtain detection results of the isolates that carry the gene which isolates number 71. alkB amplification results of 71 isolates produce ribbon DNA with size 550 bp. DNA tape with a length of 550 bp is alkB gene.The results of sequencing showed that the isolate 71 is Bordetella sp. ;Detection gene alkane monooxygenases (alkB) from marine bacteria in Pari Island Kepulauan Seribu, Jakartahas been researched. The research aims to obtain bacterial isolates that carry the gene alkBin Pari Island Kepulauan Seribu, Jakarta. The study was conducted during the five months from February 2015 to May 2015 with a method of Polymerase Chain Reaction (PCR) from 81 isolates that have been rejuvenated. Bacterial isolates rejuvenated using marine medium agar (MA) with the quadrant streak method. Obtain detection results of the isolates that carry the gene which isolates number 71. alkB amplification results of 71 isolates produce ribbon DNA with size 550 bp. DNA tape with a length of 550 bp is alkB gene.The results of sequencing showed that the isolate 71 is Bordetella sp. , Detection gene alkane monooxygenases (alkB) from marine bacteria in Pari Island Kepulauan Seribu, Jakartahas been researched. The research aims to obtain bacterial isolates that carry the gene alkBin Pari Island Kepulauan Seribu, Jakarta. The study was conducted during the five months from February 2015 to May 2015 with a method of Polymerase Chain Reaction (PCR) from 81 isolates that have been rejuvenated. Bacterial isolates rejuvenated using marine medium agar (MA) with the quadrant streak method. Obtain detection results of the isolates that carry the gene which isolates number 71. alkB amplification results of 71 isolates produce ribbon DNA with size 550 bp. DNA tape with a length of 550 bp is alkB gene.The results of sequencing showed that the isolate 71 is Bordetella sp. ]

Abstrak :
L-glutaminase (L-glutamine amidohydrolase, EC 3.5.1.2) telah menarik perhatian para peneliti karena manfaatnya dalam industri farmasi dan makanan. Bakteri laut merupakan sumber penghasil L-glutaminase yang paling diminati, terutama untuk memperoleh L-glutaminase yang tahan garam. Pada penelitian ini telah dilakukan penapisan dan karakterisasi L-glutaminase ekstraselular yang dihasilkan oleh bakteri laut dari perairan Sangihe-Talaud, Sulawesi Utara, Indonesia. Penapisan L-glutaminase secara kualitatif menggunakan media cair (Padma, et.al., 2009) dan metode pengukuran aktivitas L-glutaminase dilakukan secara spektrofotometri berdasarkan metode Imada, et.al (1973). Identifikasi isolat murni dengan aktivitas L-glutaminase paling tinggi dilakukan menggunakan sekuensing gen 16S rRNA. Terdapat 7 isolat menunjukkan hasil positif L-glutaminase, satu diantaranya dengan aktivitas 147,99 U/L atau setara dengan aktivitas spesifik 62,32 Unit/mg dipilih untuk diidentifikasi lebih lanjut. Hasil sekuensing gen 16S rRNA isolat bakteri menunjukkan kemiripan 96% dengan Pseudomonas aeruginosa strain CG-T8. Parameter fisika yang mempengaruhi produksi L-glutaminase menunjukkan produksi optimum pada suhu 30 0C, kecepatan rotasi 100 rpm, pH media 6, dan konsentrasi starter inokulum 5%. Karakterisasi aktivitas L-glutaminase ekstraselular dari Pseudomonas aeruginosa strain CG-T8 (isolat II.1) menunjukkan kondisi optimum aktivitas enzim pada suhu 37-45 0C, dan pH 7. Aktivitas enzim stabil pada penambahan larutan NaCl hingga 8% dan aktivitasnya mulai berkurang pada penambahan larutan NaCl 16% dan 20% dengan aktivitas relatif berturut-turut mencapai 79,00% dan 74,22%. Pengaruh penambahan ion-ion logam seperti Mn2+, Mg2+, dan Co2+ menunjukkan kenaikan aktivitas, sedangkan pada penambahan ion logam Zn2+, Fe3+, dan Ca2+ aktivitas enzim menurun. Bobot molekul L-glutaminase berkisar 42 kDa dan 145 kDa. ......L-glutaminase (L-glutamine amidohydrolase, EC 3.5.1.2) has attracted much attention with respect to proposed applications in both pharmaceuticals and food industries. Salt-tolerant L-glutaminase produced by marine bacteria become the most desirable in food industry. The current work details the screening of L-glutaminase producing marine bacteria from Sangihe-Talaud Sea, in North of Sulawesi, Indonesia. Screening of L-glutaminase was done using a broth medium (Padma et.al., 2009) and measurement of L-glutaminase activity carried out by spectrophotometry (Imada, et.al., 1973). Identification of selected isolate was performed by analysis of 16S rRNA gene sequence. There are seven isolates showed positive results of L-glutaminase, one of them with the activity 147.99 U/L, equivalent to the specific activity of 62.32 units / mg was selected for further study. Bacterial identification based on 16S rRNA gene sequencing has revealed the isolate 96% similarity as Pseudomonas aeruginosa strain CG-T8. Optimization of physical parameters that affect the production of L-glutaminase production showed an optimum at 30 0C, 100 rpm, pH of medium 6, and with 5% of starter inoculum. Characterization of extracellular L-glutaminase from Pseudomonas aeruginosa strain CG-T8 (II.1 isolates) showed the enzyme activity was optimum at temperature 37-45 0C, and pH 7. The enzyme activity was stable in the addition of NaCl solution up to 8% and began to decrease on addition of NaCl solution 16% and 20% with relative activity consecutively 79.00% and 74.22%. The effect of metal ions such as Mn2+, Mg2+, and Co2+ showed increased enzyme activity, whereas the addition of others metal ions (Zn2+, Fe3+, and Ca2+) decreased the activity. The molecular weights of L-glutaminase was found around 42 kDa and 145 kDa.