title: Screening, gene sequencing and characterising of lipase for methanolysis of crude palm oil.
abstract: Staphylococcus sp. WL1 lipase (LipFWS) was investigated for methanolysis of crude palm oil (CPO) at moderate temperatures. Experiments were conducted in the following order: searching for the suitable bacterium for producing lipase from activated sludge, sequencing lipase gene, identifying lipase activity, then synthesising CPO biodiesel using the enzyme. From bacterial screening, one isolated specimen which consistently showed the highest extracellular lipase activity was identified as Staphylococcus sp. WL1 possessing lipFWS (lipase gene of 2,244 bp). The LipFWS deduced was a protein of 747 amino acid residues containing an α/β hydrolase core domain with predicted triad catalytic residues to be Ser474, His704 and Asp665. Optimal conditions for the LipFWS activity were found to be at 55 °C and pH 7.0 (in phosphate buffer but not in Tris buffer). The lipase had a K(M) of 0.75 mM and a V(max) of 0.33 mMmin(-1) on p-nitrophenyl palmitate substrate. The lyophilised crude LipFWS performed as good as the commonly used catalyst potassium hydroxide for methanolysis of CPO. ESI-IT-MS spectra indicated that the CPO was converted into biodiesel, suggesting that free LipFWS is a worthy alternative for CPO biodiesel synthesis.
authors: Biochemistry Division, Faculty of Mathematics and Natural Sciences, Institut Teknologi Bandung, Jl. Ganesha 10, Bandung 40132, Indonesia.
authors: Enny
authors: E
authors: Ratnaningsih
authors: Dewi
authors: D
authors: Handayani
authors: Fatiha
authors: F
authors: Khairunnisa
authors: Ihsanawati
authors: Sari Dewi
authors: SD
authors: Kurniasih
authors: Bill
authors: B
authors: Mangindaan
authors: Sinta
authors: S
authors: Rismayani
authors: Cica
authors: C
authors: Kasipah
authors: Zeily
authors: Z
authors: Nurachman
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D055162:Biocatalysis; D056804:Biofuels; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008049:Lipase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D008697:Methane / Q000737:chemistry*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D000073878:Palm Oil; D010938:Plant Oils / Q000378:metabolism*; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D017422:Sequence Analysis, DNA; D012722:Sewage / Q000382:microbiology; D013210:Staphylococcus / Q000737:chemistry* / Q000201:enzymology; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D056804:Biofuels; D010938:Plant Oils; D012722:Sewage; D000073878:Palm Oil; D008049:Lipase; D008697:Methane