Molecular Cloning, Sequencing and Expression of an Organic Solvent-Tolerant Lipase Gene from Bacillus Sphaericus 205Y

Previously, a total of 131 organic solvent tolerant isolates originating from soil samples were successfully isolated via direct plating method using 1% (v/v) of either benzene, toluene, or a mixture of benzene and toluene as their sole carbon sources. Six out of these isolates demonstrated high...

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Main Author:	Chin, John Hun
Format:	Thesis
Language:	English English
Published:	2002
Subjects:	Organic solvents Molecular cloning
Online Access:	http://psasir.upm.edu.my/id/eprint/9518/1/FSAS_2002_47_A.pdf
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id	my-upm-ir.9518
record_format	uketd_dc
spelling	my-upm-ir.95182012-05-18T02:06:35Z Molecular Cloning, Sequencing and Expression of an Organic Solvent-Tolerant Lipase Gene from Bacillus Sphaericus 205Y 2002-06 Chin, John Hun Previously, a total of 131 organic solvent tolerant isolates originating from soil samples were successfully isolated via direct plating method using 1% (v/v) of either benzene, toluene, or a mixture of benzene and toluene as their sole carbon sources. Six out of these isolates demonstrated high tolerance of up to 75% (v/v) concentration of BTEX (benzene, toluene, ethyl benzene and xylene). These isolates were screened for lipolytic activity using triolein plates. Isolate 205y was found to produce the highest lipase yield in liquid medium and later identified as Bacillus sphaericus via 16S ribosomal DNA gene sequencing. In order to increase lipase yield, four production media were used and medium M3 yielded the highest lipase activity of 0.42 Umr⁻¹min⁻¹ at 36h incubation time. Stability studies in various organic solvents of 25% (v/v) for 30 min showed that lipase from B. sphaericus 205y was not only stable but also can be activated by hexane and xylene by 3.5 and 2.9 folds respectively. The activity, however, was greatly reduced in dimethyl sulfoxide and completely inactivated by hexadecane and acetonitrile. The gene coding for the extracellular lipase of Bacillus sphaericus 205y was isolated via genomic library and cloned into pUC 19. Organic solvents Molecular cloning 2002-06 Thesis http://psasir.upm.edu.my/id/eprint/9518/ http://psasir.upm.edu.my/id/eprint/9518/1/FSAS_2002_47_A.pdf application/pdf en public masters Universiti Putra Malaysia Organic solvents Molecular cloning Faculty of Science and Environmental Studies English
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English English
topic	Organic solvents Molecular cloning
spellingShingle	Organic solvents Molecular cloning Chin, John Hun Molecular Cloning, Sequencing and Expression of an Organic Solvent-Tolerant Lipase Gene from Bacillus Sphaericus 205Y
description	Previously, a total of 131 organic solvent tolerant isolates originating from soil samples were successfully isolated via direct plating method using 1% (v/v) of either benzene, toluene, or a mixture of benzene and toluene as their sole carbon sources. Six out of these isolates demonstrated high tolerance of up to 75% (v/v) concentration of BTEX (benzene, toluene, ethyl benzene and xylene). These isolates were screened for lipolytic activity using triolein plates. Isolate 205y was found to produce the highest lipase yield in liquid medium and later identified as Bacillus sphaericus via 16S ribosomal DNA gene sequencing. In order to increase lipase yield, four production media were used and medium M3 yielded the highest lipase activity of 0.42 Umr⁻¹min⁻¹ at 36h incubation time. Stability studies in various organic solvents of 25% (v/v) for 30 min showed that lipase from B. sphaericus 205y was not only stable but also can be activated by hexane and xylene by 3.5 and 2.9 folds respectively. The activity, however, was greatly reduced in dimethyl sulfoxide and completely inactivated by hexadecane and acetonitrile. The gene coding for the extracellular lipase of Bacillus sphaericus 205y was isolated via genomic library and cloned into pUC 19.
format	Thesis
qualification_level	Master's degree
author	Chin, John Hun
author_facet	Chin, John Hun
author_sort	Chin, John Hun
title	Molecular Cloning, Sequencing and Expression of an Organic Solvent-Tolerant Lipase Gene from Bacillus Sphaericus 205Y
title_short	Molecular Cloning, Sequencing and Expression of an Organic Solvent-Tolerant Lipase Gene from Bacillus Sphaericus 205Y
title_full	Molecular Cloning, Sequencing and Expression of an Organic Solvent-Tolerant Lipase Gene from Bacillus Sphaericus 205Y
title_fullStr	Molecular Cloning, Sequencing and Expression of an Organic Solvent-Tolerant Lipase Gene from Bacillus Sphaericus 205Y
title_full_unstemmed	Molecular Cloning, Sequencing and Expression of an Organic Solvent-Tolerant Lipase Gene from Bacillus Sphaericus 205Y
title_sort	molecular cloning, sequencing and expression of an organic solvent-tolerant lipase gene from bacillus sphaericus 205y
granting_institution	Universiti Putra Malaysia
granting_department	Faculty of Science and Environmental Studies
publishDate	2002
url	http://psasir.upm.edu.my/id/eprint/9518/1/FSAS_2002_47_A.pdf
_version_	1747810971733196800

Molecular Cloning, Sequencing and Expression of an Organic Solvent-Tolerant Lipase Gene from Bacillus Sphaericus 205Y

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