UPSI Digital Repository (UDRep)
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UPSI Digital Repository (UDRep)
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| Abstract : Perpustakaan Tuanku Bainun |
| FLL1 gene encodes an endogenous oil palm lipase that influence the innate quality of palm oil. While the genetic information is well-documented, FLL1 characteristics at the protein level remained unexplored. Therefore, this study aims to express and characterize recombinant protein FLL1 to understand its physical and chemical properties. FLL1 was efficiently expressed in Rosetta_ (DE3) pLysS competent cells using pFLL1-pGEX6P2 at 37 _C with 0.4 mM IPTG at A660 0.9. A purification method for producing high-purity FLL1 protein was established through affinity and ion exchange chromatography, suitable for downstream characterization and application. FLL1 enzyme activity peaked at 40 _C, and demonstrated its thermal stability by retaining 50 % of its activity after four hours of incubation at a similar temperature. FLL1 functioned best at pH 7 (PBS buffer) and remained highly stable at pH 8 (PBS and Tris-HCl buffers). Low LogPo/w organic solvents, especially isopropanol, significantly enhanced FLL1 activity, expanding its potential for organic solvent-rich applications. Cofactor analysis showed that Mn__ and Ca2+ are good enhancers for FLL1 activity, while Ni__ and Cu2+ showed a promising inhibitory effect, thus demonstrating the ability of certain metal ions to regulate FLL1 activity. FLL1 exhibited strong hydrolysis activity towards all substrates tested, especially palm oil, short and long fatty acid chains, highlighting its versatility for lipid processing purposes. In conclusion, FLL1 protein research findings benefit palm oil quality improvement efforts by advancing the research on oil palm lipase. The broad substrate range, metal ion dependency, and stability in organic solvents make this enzyme a promising candidate as a biocatalyst in various industries. Therefore, this study has provided a solid foundation for future FLL1 protein engineering, by contributing valuable insights into the biochemical properties of the enzyme. |
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