ISOLATION AND ENZYME ASSAY OF LIPASE AND XYLANASE ACTIVITY IN A COMPOSTING PROCESSES

• Main Author: CITRA TESALONIKA (NIM : 10508062); Pembimbing I : Fida Madayanti Warganegara, Ph.D; Pemb, ANASTASIA
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/14854
• Institution: Institut Teknologi Bandung

In a process of composting, microrganisms have a potential role as a producer of industrial enzymes such as lipase and xylanase. This study aims to isolate lipase and xylanase in the mesophilic and thermophilic phases of composting (25 oC and 49 oC, respectively) and to determine which phase of composting has the optimal enzyme activity. To achieve these goals, microbes isolated from the composting samples were screened to obtained lipase and xylanase activities and followed by identification of lipase and xylanase-producing bacteria. Finally, the activities of lipase and xylanase were determined quantitaively. Lipaseproducing <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> bacteria was screened using Luria Bertani medium containing Rhodamin B and olive oil as a substrate. Bacterial colonies were selected based on the resulting glow under UV light 350 nm. Screening xylanase-producing bacteria was performed using selective agar medium containing xylan substrate. Bacterial colonies were selected based on a clear zone. <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> <br /> Identification of lipase and xylanase-producing bacteria was carried out by a Gram staining test and 16S rDNA sequence using forward primer BacF and reverse UniB. Analysis of 16S rDNA gene was identified with the Basic Local Alignment Search Tool (BLAST) program. The result showed that the colony A1 had 85% sequence similarity with Kurthia gibsonii strain, the colony A2 had 99% sequence similarity with Providencia rettgeri strain: DSM, the colony B1 had 99% sequence similarity with Bacillus thermoamylovorans strain and the colony B2 had 98% sequence similarity with Bacillus sonorensis strain NRLL B-23154. The optimum spesific activity of lipase was observed in the colony B2 (0,0717 Unit/mg) at compost phase of 49 oC and the xylanase spesific activity was observed in the colony A2 (0,1664 Unit/mg, unit : &#956;mol/minute) at compost phase of 25 oC.