Isolasi dan identifikasi bakteri asam laktat dari limbah sawi putih

Lestari, Yuni Siti (2022) Isolasi dan identifikasi bakteri asam laktat dari limbah sawi putih. Sarjana thesis, UIN Sunan Gunung Djati Bandung.

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Abstract

INDONESIA : Limbah merupakan bahan yang dibuang dari sumber aktifitas manusia maupun alam yang belum memiliki nilai ekonomi. Limbah yang dapat dimanfaatkan dari buangan aktifitas manusia salah satu limbah pasar sayur. Limbah pasar sayur merupakan kumpulan dari berbagai macam sayuran yang tidak layak dijual. Secara fisik limbah sayur mudah busuk karena kadar air yang tinggi khususnya limbah sawi putih. Pemanfaatan limbah sawi putih adalah dengan diisolasi atau diperoleh Bakteri Asam Laktat (BAL). Bakteri Asam Laktat (BAL) merupakan bakteri anaerob fakultatif yang mampu hidup pada berbagai habitat yang cukup luas. Namun, secara ketersediaan masih sedikit dan harganya mahal. Oleh karena itu, perlu dilakukan isolasi BAL dari sumber-sumber baru seperti sawi putih yang memiliki kandungan BAL yang tinggi. Tujuan penelitian ini yaitu isolasi dan identifikasi bakteri asam laktat dari limbah sawi putih. Isolasi dilakukan untuk mendapatkan isolat bakteri asam laktat dari limbah sawi putih, kemudian dilakukan pengamatan isolat dan sel dengan pewarnaan gram. Selanjutnya diamplifikasi menggunakan instrumen Polymerase Chain Reaction (PCR) dengan primer forward BactF1 dan reverse UniB1 untuk memperoleh pragmen gen 16s rRNA dan dilakukan skuensing untuk mengetahui urutan basa nukleotidan dan dibandingkan dengan urutan basa 16s rRNA bakteri yang telah terdaftar di Bank Gen untuk diketahui homologinya dengan bakteri lain. Hasil pewarnaan gram menunjukan isolat bakteri dari limbah sawi putih sawi 1, sawi 2, sawi 3, sawi 4, sawi 5 dan sawi 6 merupakan bakteri gram positif. Hasil amplifikasi DNA kromosom isolat bakteri dari limbah sawi putih sawi 1, sawi 2 dan sawi 3 menunjukan pita DNA dengan ukuran ± 1200 bp, sehingga dapat disimpulkan bahwa proses amplifikasi fragmen gen 16s rRNA pada isolat sawi 1, sawi 2 dan sawi 3 berhasil dilakukan. Sedangkan hasil amplifikasi DNA kromosom isolat sawi 4 menunjukkan tidak adanya pita yang terseparasi karena roses amplifikasi fragmen gen 16s rRNA pada DNA sawi 4 isolat bakteri tersebut tidak berhasil. Untuk sawi 5 dan sawi 6 tidak dilanjutkan dengan amplifikasi 16s rRNA karena pada proses ekstraksi DNA kromosom tidak terdapat pita. Hasil analisis urutan basa 16s rrna menunjukan isolat sawi 1 memiliki indeks kemiripan sebesar 84,65% dengan Baccillus sp, gen isolat Sawi 2 memiliki indeks kemiripan sebesar 84,09% dengan Uncultured bacterium clone, serta gen isolat Sawi 3 memiliki indeks kemiripan sebesar 85,42% dengan Environmental 16s rDNA sequence. ENGLISH : Waste is material that is disposed of from sources of human and natural activities that do not yet have economic value. One of the wastes that can be utilized from human activities is vegetable market waste. Vegetable market waste is a collection of various kinds of vegetables that are not suitable for sale. Physically, the waste is perishable due to its high water content, especially white saw waste. The utilization of chicory waste is isolated or obtained by Lactic Acid Bacteria (LAB). Lactic Acid Bacteria (LAB) are facultative anaerobic bacteria that can live in a wide variety of habitats. However, the availability is still low and the price is expensive. Therefore, it is necessary to isolate LAB from new sources such as chicory which has a high LAB content. The purpose of this study was to isolate and isolate lactic acid bacteria from white saw waste. Isolation was carried out to obtain lactic acid isolates from eucalyptus waste, then observed the isolates and cells with gram staining. Then it was amplified using a Polymerase Chain Reaction (PCR) instrument with BactF1 and reverse UniB1 primers to obtain the 16s rRNA gene pragment and sequenced to determine the nucleotide base sequence and compared with the bacterial 16s rRNA base sequence recorded in the Gene Bank to determine its homology with bacteria. other The results of gram staining showed that the bacterial isolates from the waste of mustard 1, mustard 2, mustard 3, mustard 4, mustard 5 and mustard 6 were gram positive bacteria. The results of chromosomal DNA amplification of bacterial isolates from sawdust waste mustard 1, mustard 2 and mustard 3 showed DNA bands with a size of ± 1200 bp, so it can be said that the process of amplification of the 16s rRNA gene fragment in mustard 1, mustard 2 and mustard 3 isolates was successful. Meanwhile, the results of amplification of the chromosomal DNA of the mustard 4 isolate showed no separated bands because the process of amplification of the 16s rRNA gene fragment in the mustard 4 DNA of the bacterial isolate was not successful. For mustard 5 and mustard 6, 16s rRNA was not amplified because there was no band in the chromosomal DNA extraction process. The results of the base sequence analysis of 16s rRNA showed that the mustard 1 isolate had an index index of 84.65% with Bacillus sp, the gene for the isolate Sawi 2 had an index index of 84.09% with an Uncultured bacteria clone, and the gene isolate Mustard 3 had an index index of 85, 42% with Environmental 16s rDNA sequences.

Item Type: Thesis (Sarjana)
Uncontrolled Keywords: bakteri asam laktat; fragmen gen 16s rRNA; PCR; pewarnaan gram; skuensing.
Subjects: Biochemistry
Divisions: Fakultas Sains dan Teknologi > Program Studi Kimia
Depositing User: Yuni Siti Lestari
Date Deposited: 18 Jan 2022 04:38
Last Modified: 18 Jan 2022 04:38
URI: https://etheses.uinsgd.ac.id/id/eprint/48014

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