Kusmiyati, Nur (2013) Deteksi Lactobacillus spp. pada Susu Kambing dan Sapi Hasil Fermentasi Alami. Magister thesis, Universitas Brawijaya.
Abstract
Penelitian ini bertujuan untuk deteksi dan variabilitas Lactobacillus spp. pada susu kambing dan sapi hasil fermentasi alami. Penelitian dilaksanakan di Laboratorium Sentral Ilmu Hayati (LSIH) dan Biosain, Universitas Brawijaya, Malang pada bulan Agustus 2012Juli 2013. Penelitian ini menggunakan tiga ulangan sampel susu kambing yang berasal dari UPT Singosari dan susu sapi yang berasal dari Koperasi Sumber Sekar Laboratorium Brawijaya. Sampel susu yang diperoleh kemudian difermentasi selama 144 jam dan dilakukan isolasi bakteri. Karakter fenotipik yang digunakan sebagai parameter adalah morfologi koloni bakteri, bentuk sel bakteri, tipe gram, endospora, dan aktivitas katalase. Isolat-isolat uji diekstrak dengan metode CTAB/NACl untuk mendapatkan DNA genom. DNA genom yang diperoleh selanjutnya diamplifikasi, Denaturing Gradient Gel Electrophoresis (DGGE), dan skuensing gen 16S rRNA . Amplikon dari gen 16S rRNA kemudian disekuensing di GATC German menggunakan ABI Prism 3730 XL. Data sifat fenotipik dan pola pita DGGE dibandingkan dengan spesies acuan untuk mengetahui nama spesies. Data sekuen gen 16S rRNA dialignment dengan program ClustalX ver 1.81. Pohon filogenetik dikonstruksi menggunakan MEGA5 dengan algoritma ME, NJ, MP, dna ML. Nilai DNA relatedness menggunakan pairwise distances dan program XL. Hasil penelitian menunjukkan Lactobacillus terdeteksi pada susu kambing dan sapi hasil fermentasi alami. Berdasarkan karakter fenotipik menunjukkan sebelas isolat yang diperoleh dari susu kambing dan sapi hasil fermentasi alami merupakan genus Lactobacillus. Isolat uji C1A, C3A, G1A, dan G3A adalah L.plantarum karena memiliki migrasi pola pita DGGE yang sama sedangkan G3B adalah L.casei . Hasil analisis sekuen gen 16S rRNA menunjukkan sebelas isolat uji merupakan anggota Lactobacillus yang berbeda. Isolat uji C1A, C3A, G1A, dan G3A teridentifikasi sebagai L.plantarum . Isolat uji C2B, G1B, dan G2A teridentifikasi sebagai L.rhamnosus . Isolat uji C1B, C3B, dan G2B teridentifikasi sebagai L.zeae dan isolat uji G3B teridentifikasi sebagai L.casei. Identifikasi diperoleh dari program BLAST database GenBank dan MEGA5 berkorelasi dengan yang diperoleh oleh DGGE. Variabilitas Lactobacillus pada susu kambing hasil fermentasi alami yaitu L.plantarum , L.rhamnosus , L.zeae , dan L.casei sedangkan pada susu sapi yaitu L.plantarum , L.rhamnosus , dan L.zeae .
English Abstract
This research aims to detect and variability of Lactobacillus spp. in natural fermented goat and cow milk. This research has being done in Central Laboratory of Life Science and Bioscience, Brawijaya University, Malang on August 2012-July 2013. This research was using three repetition sample of goat milk from UPT Singosari and cow milk sample from Sumber Sekar Cooperation of Brawijaya Laboratory. Milk sample acquired then being fermented for 144 hours and then had the bacteria isolated. Phenotypic characteristics had identified by bacteria colony morphology, shape of bacteria cell, Gram type, endospore, and catalase activity. Tested strains were extracted with CTAB/NaCl method in order to acquire DNA genome. DNA genome acquired then being amplified in order to acquire amplicon of Denaturing Gradient Gel Electrophoresis (DGGE), and 16S rDNA sequence. Amplicon of 16S rDNA then had sequencing in Germany GATC using ABI Prism 3730 XL. Phenotypic characteristic data and DGGE band pattern was compared with reference species in order to know the species name. Data of 16S rDNA sequence was being aligned with ClustalX ver 1.81 programs. Phylogenetic tree was constructed by using MEGA5 with ME, NJ, MP, and ML algorithm. DNA value relatedness is using pair wise distances and XL program. Results showed Lactobacillus detected in goat and cow milk fermented naturally. Based on phenotypic characters showed eleven isolates obtained from goat and cow milk fermented naturally constitute the genus Lactobacillus. Isolate test C1A, C3A, G1A, and G3a is L.plantarum migration because it has the same DGGE band patterns while G3B is L.casei . The results of 16S rRNA gene sequence analysis showed eleven test isolates are members of different Lactobacillus. Isolate test C1A, C3A, G1A, and G3a identified as L.plantarum. Isolate test C2B, G1B, and G2A identified as L.rhamnosus . Isolate test C1B, C3B, and G2B and isolates identified as L.zeae G3B identified as L.casei test. Identification obtained from the GenBank database and the BLAST program MEGA5 correlated with those obtained by DGGE. Variability Lactobacillus in fermented goat milk is naturally L.plantarum , L.rhamnosus, L.zeae , and L.casei while in cows milk that is L.plantarum , L.rhamnosus , and L.zeae .
| Item Type: | Thesis (Magister) |
|---|---|
| Identification Number: | TES/579.37/KUS/d/041307539 |
| Subjects: | 500 Natural sciences and mathematics > 579 Natural history of microorganisms, fungi, algae > 579.3 Prokaryotes |
| Divisions: | S2/S3 > Magister Biologi, Fakultas MIPA |
| Depositing User: | Endro Setyobudi |
| Date Deposited: | 03 Feb 2014 14:42 |
| Last Modified: | 03 Feb 2014 14:42 |
| URI: | http://repository.ub.ac.id/id/eprint/157736 |
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