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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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 |