ISOLATION AND IDENTIFICATION THERMOPHILIC BACTERIA PRODUCING LIPASE FROM DOMESTIC WASTE

• Main Author: SUTANTO, MEGA
• Format: Final Project
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/71470
• Institution: Institut Teknologi Bandung

Thermophilic bacteria have been rapidly studied since its function in biotechnological industry to produce stable enzyme in high temperature. Lipase is one of the enzyme which is largely applied to detergent, food, cosmetics, and other industries. Thermophilic bacteria have a lot advantages compared to mesophilic bacteria since its survival to board range of temperature. Four thermophilic bacteria producing lipase were isolated from domestic organic waste. The isolates could grow at range temperature at 37 ºC to 65 ºC. The isolates have been characterized with optical density measurement and gram staining. Optical density measurement showed that all isolates had growth rate value between 0.04 to 0.1 OD h-1 (Optical Density per hour) and maximum growth at about (A600) 0.4 to 1.1 OD. After being stained, all isolates showed as gram positive bacteria and coccus formed. These isolates have been characterized further by isolating their chromosomal DNA with Klijn method and the 16S rRNA genes have been amplified with PCR. Two of them were amplified succesfully with predenaturation step at 94 ºC for 5 minutes, denaturation step 50 ºC for 1 minute, elongation step 72 ºC for 10 minutes. Agarose gel electrophoresis showed that the size of band at around 1.6 kbl. One of the 16S rRNA genes has been sequenced with Uni B1 and Bact F1 primers.The result of 16S rRNA sequence showed that the isolate was closely related to Bacillus licheniformis TKW 3 (Accession number gi|50080682|gb|AY631057.1) following comparisons to the gene bank.|.