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

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...

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Main Author:	Lee, Yuan Chern
Format:	Thesis
Language:	English English
Published:	2006
Subjects:	Monoterpenes - Enzyme activation
Online Access:	http://psasir.upm.edu.my/id/eprint/504/1/600363_fbsb_2006_20_abstrak_je__dh_pdf_.pdf
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spelling	my-upm-ir.5042013-05-27T06:48:48Z Partial Purification and Characterization of a Monoterpene Synthase Extracted from Young Leaves of Michellia Alba 2006-02 Lee, Yuan Chern 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. Monoterpenes - Enzyme activation 2006-02 Thesis http://psasir.upm.edu.my/id/eprint/504/ http://psasir.upm.edu.my/id/eprint/504/1/600363_fbsb_2006_20_abstrak_je__dh_pdf_.pdf application/pdf en public masters Universiti Putra Malaysia Monoterpenes - Enzyme activation Faculty of Biotechnology and Biomolecular Sciences English
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English English
topic	Monoterpenes - Enzyme activation
spellingShingle	Monoterpenes - Enzyme activation Lee, Yuan Chern Partial Purification and Characterization of a Monoterpene Synthase Extracted from Young Leaves of Michellia Alba
description	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.
format	Thesis
qualification_level	Master's degree
author	Lee, Yuan Chern
author_facet	Lee, Yuan Chern
author_sort	Lee, Yuan Chern
title	Partial Purification and Characterization of a Monoterpene Synthase Extracted from Young Leaves of Michellia Alba
title_short	Partial Purification and Characterization of a Monoterpene Synthase Extracted from Young Leaves of Michellia Alba
title_full	Partial Purification and Characterization of a Monoterpene Synthase Extracted from Young Leaves of Michellia Alba
title_fullStr	Partial Purification and Characterization of a Monoterpene Synthase Extracted from Young Leaves of Michellia Alba
title_full_unstemmed	Partial Purification and Characterization of a Monoterpene Synthase Extracted from Young Leaves of Michellia Alba
title_sort	partial purification and characterization of a monoterpene synthase extracted from young leaves of michellia alba
granting_institution	Universiti Putra Malaysia
granting_department	Faculty of Biotechnology and Biomolecular Sciences
publishDate	2006
url	http://psasir.upm.edu.my/id/eprint/504/1/600363_fbsb_2006_20_abstrak_je__dh_pdf_.pdf
_version_	1747810236446539776

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

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