ISOLASI DAN IDENTIFIKASI BAKTERI PROTEOLITIK PADA RUSIP UDANG WINDU (Penaeus monodon) PASCA FERMNTASI 144 JAM BERDASARKAN SEKUENS GEN 16S rRNA

HUMU,SARFINA, G1C218017 and Ethica, stalis norma, . and Sulityaningtyas,Ayu Rahmawati, . (2019) ISOLASI DAN IDENTIFIKASI BAKTERI PROTEOLITIK PADA RUSIP UDANG WINDU (Penaeus monodon) PASCA FERMNTASI 144 JAM BERDASARKAN SEKUENS GEN 16S rRNA. Sarjana / Sarjana Terapan (S1/D4) thesis, ["eprint_fieldopt_institution_Universitas Muhammadiyah Semarang" not defined].

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Abstract

ABSTRAK Kebutuhan enzim protease di Indonesia semakin meningkat. Hal ini dilihat dari berbagai industri pengolahan pangan yang memanfaatkan enzim protease. Namun, Indonesia belum mampu memproduksi enzim tersebut dalam skala besar. Tujuan penelitian ini mengisolasi adanya bakteri penghasil enzim protease yang terdapat pada rusip udang windu pasca fermentasi 144 jam, serta untuk mengetahui jenis bakteri penghasil enzim protease berdasarkan analisis gen 16S rRNA. Isolasi dan identifikasi bakteri dilakukan menggunakan media Nutrient Agar , pewarnaan gram dan uji produksi enzim protease dengan menggunakan Skim Milk Agar (SMA). Identifkasi molekuler berdasarkan analisis sekuen gen 16S rRNA yang dilakukan dengan teknik Polymerase Chain Reaction (PCR) dilanjutkan dengan sekuensing. Dari proses isolasi diperoleh hasil berupa isolat bakteri yang meiliki aktivitas proteolitik yang baik yang ditandai dengan adanya zona bening pda media Susu Skim Agar yang memiliki diameter sebesar 4mm yaitu pada isolat RPM.BBSH. Berdasarkan analisis sekuen gen 16S rRNA menunjukkan bahwa isolat bakteri RPM.BBSH memiliki kemiripan 100% dengan fragmen gen 16S rRNA dengan isolat bakteri Blast DNA Baser Staphylococus warneri strain SR4-3 (Genbank kode akses: MN421509.1). Berdasarkan hasil penelitian ini dapat disimpulkan bahwa isolate RPM.BBSH berpotensi sebagai penghasil enzim protease dan berdasarkan identifikasi molekuler yang dilakukan isolat RPM.BBSH dinyatakan sebagai Staphylococcus warneri IRPMD ( Indonesia Rusip Penaeus monodon Day-6). Kata Kunci: Identifikasi Molekuler, bakteri proteolitik, gen 16S rRNA Staphylococcus warneri Isolation and identification of Proteolytic Bacteria in Rusip Shrimp Windu (Penaeus monodon) Post Fermentation 144 Hours Based on 16S rRNA gene sequences Sarfina Humu1, Ayu Rahmawati Sulistyaningtyas2, Stalis Norma Ethica3 1. D-IV Study Program Health Analyst, Faculty of Nursing and Health, University of Muhammadiyah Semarang 2. 2. D-III Study Program Health Analyst, Faculty of Nursing and Health, University of Muhammadiyah Semarang 3. Master of Science Study Program Faculty of Health and Nursing, University of Muhammadiyah Semarang ABSTRACT The need for protease enzymes in Indonesia is increasing. This is seen from a variety of food processing industries that utilize protease enzymes. However, Indonesia has not been able to produce these enzymes on a large scale. The purpose of this study was to determine the presence of protease-producing bacteria found in the rusip of tiger shrimp after 144 hours fermentation, as well as to determine the types of bacteria that produce protease enzymes based on 16S rRNA gene analysis. Isolation and identification of bacteria was carried out using Agar Nutrient media, gram staining and protease enzyme production test using Skim Milk Agar (SMA). Molecular identification is based on 16S rRNA gene sequence analysis using Polymerase Chain Reaction (PCR) technique using agarose gel and continued with sequencing. The isolation process results in the form of bacterial isolates which have good proteolytic activity which is marked by the presence of a clear zone in the Skim Milk media agar which has a diameter of 4mm, that is the RPM.BBSH isolate. Based on the analysis of 16S rRNA gene sequences, it showed that RPM BBSH isolates had 100% similarity with 16S rRNA gene fragments with Staphylococus warneri strain SR4-3 isolates (Genbank kode akses: MN421509.1). Based on the results of this study it can be concluded that the RPM.BBSH isolate has the potential to produce protease enzymes and based on the molecular identification of the RPM.BBSH isolates are expressed as Staphylococcus warneri IRPMD ( Indonesia Rusip Penaeus monodon Day-6). Key words: Molecular Identification, Proteolytic Bacteria, 16S rRNA gen Staphylococcus warneri

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