Characterization Of Variants Of Polyhydroxyalkanoate Synthase Of Cupriavidus Necator Generated By Error-Prone Polymerase Chain Reaction

In this study, error-prone PCR of type I polyhydroxyalkanoate synthase (PhaCCn) from Cupriavidus necator was carried out to introduce random mutations in PhaCCn to investigate the functional importance of the substituted amino acids of PhaCCn, which to date, is structurally unknown.. Via this approa...

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Main Author:	Loh, Keat Lay
Format:	Thesis
Language:	English
Published:	2013
Subjects:	QH301 Biology
Online Access:	http://eprints.usm.my/45111/1/Loh%20Keat%20Lay24.pdf
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id	my-usm-ep.45111
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spelling	my-usm-ep.451112019-07-31T08:23:32Z Characterization Of Variants Of Polyhydroxyalkanoate Synthase Of Cupriavidus Necator Generated By Error-Prone Polymerase Chain Reaction 2013-07 Loh, Keat Lay QH301 Biology In this study, error-prone PCR of type I polyhydroxyalkanoate synthase (PhaCCn) from Cupriavidus necator was carried out to introduce random mutations in PhaCCn to investigate the functional importance of the substituted amino acids of PhaCCn, which to date, is structurally unknown.. Via this approach, mutant H8, which harboured five amino acid substitutions (I71T, I172T, P339L, N514S and S522R) was obtained. The H8 mutant exhibited low P(3HB) accumulation (0.1% CDW (Cell dry weight)) when compared to the wild-type (11.3% CDW). In terms of their positions in PhaCCn sequence, mutation I71T is located at the highly variable N-terminal domain of the PhaCCn, the mutation I172T is located in between the N-terminal and the conserved region, while mutations N514S, P339L and S522R were located in the α/β hydrolase fold region, which is highly conserved in all classes of PHA synthases. 2013-07 Thesis http://eprints.usm.my/45111/ http://eprints.usm.my/45111/1/Loh%20Keat%20Lay24.pdf application/pdf en public masters Universiti Sains Malaysia Pusat Pengajian Sains Kajihayat
institution	Universiti Sains Malaysia
collection	USM Institutional Repository
language	English
topic	QH301 Biology
spellingShingle	QH301 Biology Loh, Keat Lay Characterization Of Variants Of Polyhydroxyalkanoate Synthase Of Cupriavidus Necator Generated By Error-Prone Polymerase Chain Reaction
description	In this study, error-prone PCR of type I polyhydroxyalkanoate synthase (PhaCCn) from Cupriavidus necator was carried out to introduce random mutations in PhaCCn to investigate the functional importance of the substituted amino acids of PhaCCn, which to date, is structurally unknown.. Via this approach, mutant H8, which harboured five amino acid substitutions (I71T, I172T, P339L, N514S and S522R) was obtained. The H8 mutant exhibited low P(3HB) accumulation (0.1% CDW (Cell dry weight)) when compared to the wild-type (11.3% CDW). In terms of their positions in PhaCCn sequence, mutation I71T is located at the highly variable N-terminal domain of the PhaCCn, the mutation I172T is located in between the N-terminal and the conserved region, while mutations N514S, P339L and S522R were located in the α/β hydrolase fold region, which is highly conserved in all classes of PHA synthases.
format	Thesis
qualification_level	Master's degree
author	Loh, Keat Lay
author_facet	Loh, Keat Lay
author_sort	Loh, Keat Lay
title	Characterization Of Variants Of Polyhydroxyalkanoate Synthase Of Cupriavidus Necator Generated By Error-Prone Polymerase Chain Reaction
title_short	Characterization Of Variants Of Polyhydroxyalkanoate Synthase Of Cupriavidus Necator Generated By Error-Prone Polymerase Chain Reaction
title_full	Characterization Of Variants Of Polyhydroxyalkanoate Synthase Of Cupriavidus Necator Generated By Error-Prone Polymerase Chain Reaction
title_fullStr	Characterization Of Variants Of Polyhydroxyalkanoate Synthase Of Cupriavidus Necator Generated By Error-Prone Polymerase Chain Reaction
title_full_unstemmed	Characterization Of Variants Of Polyhydroxyalkanoate Synthase Of Cupriavidus Necator Generated By Error-Prone Polymerase Chain Reaction
title_sort	characterization of variants of polyhydroxyalkanoate synthase of cupriavidus necator generated by error-prone polymerase chain reaction
granting_institution	Universiti Sains Malaysia
granting_department	Pusat Pengajian Sains Kajihayat
publishDate	2013
url	http://eprints.usm.my/45111/1/Loh%20Keat%20Lay24.pdf
_version_	1747821456843079680

Characterization Of Variants Of Polyhydroxyalkanoate Synthase Of Cupriavidus Necator Generated By Error-Prone Polymerase Chain Reaction

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