Partial Purification and Characterization of a Monoterpene Synthase Extracted from Young Leaves of Michellia Alba

Lee, Yuan Chern (2006) Partial Purification and Characterization of a Monoterpene Synthase Extracted from Young Leaves of Michellia Alba. Masters thesis, Universiti Putra Malaysia.

[img] PDF
119Kb

Abstract

This study was conducted to partially purify linalool synthase from the young leaves of Michellia alba (Cempaka Putih) from the Magnoliaceae family. The technique used to determine the amount of linalool produced from enzyme activity was the combination of solid-phase microextraction (SPME) and gas chromatography with flame-ionization detecter (GCFID) technique. The substrate used for this enzyme activity assay was geranylpyrophosphate (GPP). Optimal conditions such as temperature and incubation time for SPME technique were also determined. The linalool synthase exhibited a strict requirement for a divalent metal cofactor with a preference for Mg2+, Mn2+ and K+ ions. The optimal pH and temperature of the enzyme was 6.0 and 30oC respectively. The enzyme was inhibited by 1,2-Di(2-aminoethoxy)ethane-N,N,N’N’-tetra-acetic acids (EGTA). Three steps of partial purification of enzyme were carried out, including the crude extraction of young leaves, ultra centrifugation and Mono-Q anion exchange chromatography. The partially purified linalool synthase was characterized and studied for its enzyme kinetic properties. The linalool synthase has a Km of 83µM for substrate GPP. The SPME-GCFID technique was later proved to be reliable and sensitive in determination of monoterpene products.

Item Type:Thesis (Masters)
Subject:Monoterpenes - Enzyme activation
Chairman Supervisor:Associate Professor Radzali Muse, PhD
Call Number:FBSB 2006 20
Faculty or Institute:Faculty of Biotechnology and Biomolecular Sciences
ID Code:504
Deposited By: Yusfauhannum Mohd Yunus
Deposited On:14 Oct 2008 14:42
Last Modified:27 May 2013 06:48

Repository Staff Only: item control page

Document Download Statistics

This item has been downloaded for since 14 Oct 2008 14:42.

View statistics for "Partial Purification and Characterization of a Monoterpene Synthase Extracted from Young Leaves of Michellia Alba"


Universiti Putra Malaysia Institutional Repository

Universiti Putra Malaysia Institutional Repository is an on-line digital archive that serves as a central collection and storage of scientific information and research at the Universiti Putra Malaysia.

Currently, the collections deposited in the IR consists of Master and PhD theses, Master and PhD Project Report, Journal Articles, Journal Bulletins, Conference Papers, UPM News, Newspaper Cuttings, Patents and Inaugural Lectures.

As the policy of the university does not permit users to view thesis in full text, access is only given to the first 24 pages only.