ISOLATION AND DETERMINATIAN OF LIPASE GENE SEQUENCE FROM STAPHYLOCOCCUS WL1 ISOLATE

• Main Author: KHAIRUNNISA (NIM : 10508054); Pembimbing : Zeily Nurachman, D.Sc , FATIHA
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/15648
• Institution: Institut Teknologi Bandung

Staphylococcus WL1 isolate that is obtained from previous research produces lipase for catalyzing transesterification of crude palm oil into biodiesel. To obtain large amount of lipase, first searching on Staphylococcus WL1 lipase gene should be performed. In the present study, steps for the research included designing specific primers for lipase gene, amplification of lipase gene fragment using PCR, cloning lipase gene fragment into pGEM®-T Easy plasmid, and determination of nucleotides sequence of the gene. The result showed that amplification of the Staphylococcus WL1 chromosomal DNA using primers: forward primer (5'-GAG GTG CCG AAT ATG AAA AAT AGA G-3') and reverse primer (5'-TTA TGC TTG TGC TTT TTG CTC TG-3') produced a nucleotide fragment of 2,244 bp. This was a gene encoding lipase FWS (called lipFWS) including ATG as a start codon and TAA as a stop codon. Furthermore, the gene showed 96% identity to lipase genes of Staphylococcus warneri strain gehWC (GeneBank No. AB189474.1). In silico translation analysis showed that LipFWS consisted of 747 amino acid residues. The enzyme belongs to α/β hydrolase family with predicted catalytic residues at Ser474, Asp665, and His704.