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 psychrophili...
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Pusat Pengajian Sains Perubatan Universiti Sains Malaysia
2008
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ftunivsainsmal:oai:generic.eprints.org:50393 2023-05-15T14:00:58+02:00 Expression and purification of glyceraldehyde-3-phosphate dehydrogenase from psychrophilic bacterium Few Ling, Ling 2008 application/pdf http://eprints.usm.my/50393/ http://eprints.usm.my/50393/1/DR.%20FEW%20LING%20LING%20-%2024%20pages.pdf en eng Pusat Pengajian Sains Perubatan Universiti Sains Malaysia 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) R Medicine (General) Article PeerReviewed 2008 ftunivsainsmal 2021-11-02T18:14:52Z 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. Article in Journal/Newspaper Antarc* Antarctic eprints@USM (Universiti Sains Malaysia) Antarctic The Antarctic |
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R Medicine (General) |
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R Medicine (General) Few Ling, Ling Expression and purification of glyceraldehyde-3-phosphate dehydrogenase from psychrophilic bacterium |
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R Medicine (General) |
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. |
format |
Article in Journal/Newspaper |
author |
Few Ling, Ling |
author_facet |
Few Ling, Ling |
author_sort |
Few Ling, Ling |
title |
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_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_sort |
expression and purification of glyceraldehyde-3-phosphate dehydrogenase from psychrophilic bacterium |
publisher |
Pusat Pengajian Sains Perubatan Universiti Sains Malaysia |
publishDate |
2008 |
url |
http://eprints.usm.my/50393/ http://eprints.usm.my/50393/1/DR.%20FEW%20LING%20LING%20-%2024%20pages.pdf |
geographic |
Antarctic The Antarctic |
geographic_facet |
Antarctic The Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_relation |
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) |
_version_ |
1766270366924144640 |