Expression and purification of glyceraldehyde-3-phosphate dehydrogenase from psychrophilic bacterium
Organisms that thrive in cold environments are known as psychrophiles. One of the strategies for their cold adaptation is the ability to synthesize cold-adapted enzymes. Our collection of cold-tolerant microorganisms isolated· from the Antarctic region has offered a potential source for psychroph...
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| Format: | Article |
| Language: | English |
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Pusat Pengajian Sains Perubatan Universiti Sains Malaysia
2008
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| Online Access: | http://eprints.usm.my/50393/1/DR.%20FEW%20LING%20LING%20-%2024%20pages.pdf |
| _version_ | 1797012415250956288 |
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| author | Few Ling, Ling |
| author_facet | Few Ling, Ling |
| author_sort | Few Ling, Ling |
| collection | USM |
| description | Organisms that thrive in cold environments are known as psychrophiles. One of the
strategies for their cold adaptation is the ability to synthesize cold-adapted enzymes.
Our collection of cold-tolerant microorganisms isolated· from the Antarctic region has
offered a potential source for psychrophilic enzymes. Previously our group had
successfully cloned the open reading frame for GAPDH gene from an Antarctical
bacterium known as phi9. The ORF was cloned into a pET-14b plasmid. The full
length GAPDH protein was subsequently expressed in E. coli strain BL21 (DE3),
purified as His-tag protein and confirmed to be catalytically active. Results showed
that IPTG concentration did not have any effect on protein expression and solubility
while 3 hours of induction time at room temperature (28°C) was the best conditions for
the expression and solubility of this protein. This protein was shown to be most active
at 38°C and its specific activity increased by 40% from 3.6 JlmOI/min/mg to 6.1
J.UllOifmin/mg when the temperature increased from 23°C to 38°C. This work laid the
foundation for further biochemical and structural characterizations of GAPDH from a
psychrophilic bacterium by providing a highly purified recombinant protein sample. |
| first_indexed | 2024-03-06T15:47:21Z |
| format | Article |
| id | usm.eprints-50393 |
| institution | Universiti Sains Malaysia |
| language | English |
| last_indexed | 2024-03-06T15:47:21Z |
| publishDate | 2008 |
| publisher | Pusat Pengajian Sains Perubatan Universiti Sains Malaysia |
| record_format | dspace |
| spelling | usm.eprints-503932021-10-26T02:33:54Z http://eprints.usm.my/50393/ Expression and purification of glyceraldehyde-3-phosphate dehydrogenase from psychrophilic bacterium Few Ling, Ling R Medicine (General) Organisms that thrive in cold environments are known as psychrophiles. One of the strategies for their cold adaptation is the ability to synthesize cold-adapted enzymes. Our collection of cold-tolerant microorganisms isolated· from the Antarctic region has offered a potential source for psychrophilic enzymes. Previously our group had successfully cloned the open reading frame for GAPDH gene from an Antarctical bacterium known as phi9. The ORF was cloned into a pET-14b plasmid. The full length GAPDH protein was subsequently expressed in E. coli strain BL21 (DE3), purified as His-tag protein and confirmed to be catalytically active. Results showed that IPTG concentration did not have any effect on protein expression and solubility while 3 hours of induction time at room temperature (28°C) was the best conditions for the expression and solubility of this protein. This protein was shown to be most active at 38°C and its specific activity increased by 40% from 3.6 JlmOI/min/mg to 6.1 J.UllOifmin/mg when the temperature increased from 23°C to 38°C. This work laid the foundation for further biochemical and structural characterizations of GAPDH from a psychrophilic bacterium by providing a highly purified recombinant protein sample. Pusat Pengajian Sains Perubatan Universiti Sains Malaysia 2008 Article PeerReviewed application/pdf en http://eprints.usm.my/50393/1/DR.%20FEW%20LING%20LING%20-%2024%20pages.pdf Few Ling, Ling (2008) Expression and purification of glyceraldehyde-3-phosphate dehydrogenase from psychrophilic bacterium. Universiti Sains Malaysia. (Submitted) |
| spellingShingle | R Medicine (General) Few Ling, Ling Expression and purification of glyceraldehyde-3-phosphate dehydrogenase from psychrophilic bacterium |
| title | Expression and purification of glyceraldehyde-3-phosphate
dehydrogenase from psychrophilic bacterium |
| title_full | Expression and purification of glyceraldehyde-3-phosphate
dehydrogenase from psychrophilic bacterium |
| title_fullStr | Expression and purification of glyceraldehyde-3-phosphate
dehydrogenase from psychrophilic bacterium |
| title_full_unstemmed | Expression and purification of glyceraldehyde-3-phosphate
dehydrogenase from psychrophilic bacterium |
| title_short | Expression and purification of glyceraldehyde-3-phosphate
dehydrogenase from psychrophilic bacterium |
| title_sort | expression and purification of glyceraldehyde 3 phosphate dehydrogenase from psychrophilic bacterium |
| topic | R Medicine (General) |
| url | http://eprints.usm.my/50393/1/DR.%20FEW%20LING%20LING%20-%2024%20pages.pdf |
| work_keys_str_mv | AT fewlingling expressionandpurificationofglyceraldehyde3phosphatedehydrogenasefrompsychrophilicbacterium |