Evaluasi kemampuan isolat bakteri tanah sebagai agen biogrouting

Sumendap, Tracy Anastasia Lazialitta (2024) Evaluasi kemampuan isolat bakteri tanah sebagai agen biogrouting. Bachelor thesis, Universitas Pelita Harapan.

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Abstract

Biogrouting merupakan metode alternatif yang digunakan untuk mengurangi dampak negatif penggunaan grout kimia terhadap lingkungan. Teknik biogruting dapat dilakukan dengan memanfaatkan bakteri penghasil urease yang dapat membentuk kristal kalsium karbonat. Pada penelitian ini dilakukan identifikasi dan karakterisasi isolat NC7, LK3 dan TG6 yang berasal dari tanah di Indonesia, serta evaluasi potensi ketiga isolat sebagai agen biogrouting. Metode penelitian ini mencakup analisis morfologi; identifikasi molekuler berdasarkan sekuens 16S- rRNA; uji aktivitas urease; uji presipitasi kalsium karbonat; uji pertumbuhan bakteri pada berbagai pH; dan uji pembentukan presipitat pada berbagai pH. Ketiga isolat memiliki koloni berwarna putih, namun koloni NC7 berukuran sedang, berbentuk irregular, tekstur smooth, elevasi convex dan margin wavy; koloni LK3 memiliki ukuran kecil, tekstur smooth, berbentuk circular, elevasi raised dan margin entire; sedangkan koloni TG6 memiliki ukuran kecil, tekstur smooth sedikit lengket, berbentuk circular, elevasi raised dan margin entire. NC7 dan LK3 merupakan bakteri berbentuk batang Gram-positif yang dapat menghasilkan endospora, sedangkan TG6 merupakan bakteri berbentuk batang Gram-negatif yang tidak dapat menghasilkan endospora. Analisis sekuens 16S-rRNA menunjukkan bahwa NC7 merupakan Bacillus cereus, LK3 merupakan Rummeliibacillus sp. dan TG6 merupakan Comamonas sp. Semua isolat menunjukkan aktivitas urease, Comamonas sp. TG6 memiliki aktivitas enzim urease tertinggi. Bacillus cereus NC7 dan Comamonas sp TG6 mampu menghasilkan kristal pada medium B4 agar setelah 14 hari inkubasi, namun tidak teramati adanya kristal yang terbentuk pada kultur Rummeliibacillus sp. LK3. Pada medium B4 cair, laju pembentukan kristal dari NC7, TG6, dan LK3 adalah 0,13; 0,11; dan 0,09 g/minggu. Bacillus cereus NC7, Rummeliibacillus sp. LK3 dan Comamonas sp. TG6 tidak menunjukkan pertumbuhan pada pH 2, namun ketiganya dapat tumbuh pada pH 5, 7 dan 9. Uji presipitasi CaCO3 pada berbagai pH menunjukkan bahwa kultur Bacillus cereus NC7, Rummeliibacillus sp. LK3 dan Comamonas sp. TG6 mengalami peningkatan jumlah presipitat, seiring dengan peningkatan nilai pH dan nilai OD600 selama masa inkubasi 14 hari. Berdasarkan hasil penelitian ini, ketiga bakteri memiliki potensi sebagai agen biogrouting./Biogrouting is an alternative method used to reduce negative impact of using chemical grout on the.The biogrouting technique can be carried out by utilizing urease-producing bacteria which can from calcium carbonate crystals. In this study, the identifications and characterization of isolates NC7, LK3 and TG6 originating from soil in Indonesia were carried out, as well as evaluating the potential of the third isolate as a biogrouting agent. The research methods included morphological analysis; moleculer identification based on 16S-rRNA sequencing; urease activity test; calcium carbonate precipitation test;, bacterial growth test at various pH; and precipitate formation test at various pH. The third isolate had white colonies, but NC7 colonies was medium size, irregular shape, smooth texture, convex elevation and wavy margin; LK3 colonies have small size, smooth texture, circular shape, raised elevation and entire margin; while TG6 colonies was small size, wmooth, slightly sticky texture, circular in shape, raised elevation and entire margin. NC7 and LK3 is Gram-positive rod-shaped bacteria that can forming-endospores, while TG6 is Gram-negative rod-shaped bacteria and non-endospore-forming; The 16S rRNA sequence analysis revealed that NC7 identical to Bacillus cereus, LK3 identical to Rummeliibacillus sp. and TG6 identical to Comamonas sp.; All isolates showed urease activity, Comamonas sp. TG6 has the highest urease enzyme activity. Bacillus cereus NC7 and Comamonas sp. TG6 were able to produce calcium carbonate crystals on B4 agar medium after 14 days of incubation, but no crystals were observed in Rummeliibacillus sp. LK3 culture. In B4 broth medium, the rate of crystal formation of NC7, TG6, and LK3 was 0,13; 0,11; and 0,09 g/week. Bacillus cereus NC7, Rummeliibacillus sp. LK3 and Comamonas sp. TG6 did not who growth at pH 2, however all three could grow at pH 5, 7 and 9. CaCO3 precipitation test at various pH showed that Bacillus cereus NC7, Rummeliibacillus sp. LK3 and Comamonas sp. TG6 experienced an increase in the amount precipitate, along with an increase in pH and OD600 values during the 14 day incubation period. Based on the result this result, the third bacteria have as potential as a biogoruting agent.

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