Enzyme Sequence and Its Relationship to Hyperbaric Stability of Artificial and Natural Fish Lactate Dehydrogenases
The cDNAs of lactate dehydrogenase b (LDH-b) from both deep-sea and shallow living fish species, Corphaenoides armatus and Gadus morhua respectively, have been isolated, sequenced and their encoded products overproduced as recombinant enzymes in E. coli. The proteins were characterised in terms of t...
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ftucl:oai:eprints.ucl.ac.uk.OAI2:123018 2023-12-24T10:16:52+01:00 Enzyme Sequence and Its Relationship to Hyperbaric Stability of Artificial and Natural Fish Lactate Dehydrogenases Brindley, AA Pickersgill, RW Partridge, JC Dunstan, DJ Hunt, DM Warren, MJ 2008-04-30 application/pdf https://discovery.ucl.ac.uk/id/eprint/123018/1/123018.pdf https://discovery.ucl.ac.uk/id/eprint/123018/ eng eng PUBLIC LIBRARY SCIENCE https://discovery.ucl.ac.uk/id/eprint/123018/1/123018.pdf https://discovery.ucl.ac.uk/id/eprint/123018/ open PLOS ONE , 3 (4) , Article e2042. (2008) Article 2008 ftucl 2023-11-27T13:07:39Z The cDNAs of lactate dehydrogenase b (LDH-b) from both deep-sea and shallow living fish species, Corphaenoides armatus and Gadus morhua respectively, have been isolated, sequenced and their encoded products overproduced as recombinant enzymes in E. coli. The proteins were characterised in terms of their kinetic and physical properties and their ability to withstand high pressures. Although the two proteins are very similar in terms of their primary structure, only 21 differences at the amino acid level exist between them, the enzyme from the deep-sea species has a significantly increased tolerance to pressure and a higher thermostability. It was possible to investigate whether the changes in the N-terminal or C-terminal regions played a greater role in barophilic adaptation by the construction of two chimeric enzymes by use of a common restriction site within the cDNAs. One of these hybrids was found to have even greater pressure stability than the recombinant enzyme from the deep-living fish species. It was possible to conclude that the major adaptive changes to pressure tolerance must be located in the N-terminal region of the protein. The types of changes that are found and their spatial location within the protein structure are discussed. An analysis of the kinetic parameters of the enzymes suggests that there is clearly a trade off between Km and kcat values, which likely reflects the necessity of the deep-sea enzyme to operate at low temperatures. Article in Journal/Newspaper Gadus morhua University College London: UCL Discovery |
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University College London: UCL Discovery |
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ftucl |
language |
English |
description |
The cDNAs of lactate dehydrogenase b (LDH-b) from both deep-sea and shallow living fish species, Corphaenoides armatus and Gadus morhua respectively, have been isolated, sequenced and their encoded products overproduced as recombinant enzymes in E. coli. The proteins were characterised in terms of their kinetic and physical properties and their ability to withstand high pressures. Although the two proteins are very similar in terms of their primary structure, only 21 differences at the amino acid level exist between them, the enzyme from the deep-sea species has a significantly increased tolerance to pressure and a higher thermostability. It was possible to investigate whether the changes in the N-terminal or C-terminal regions played a greater role in barophilic adaptation by the construction of two chimeric enzymes by use of a common restriction site within the cDNAs. One of these hybrids was found to have even greater pressure stability than the recombinant enzyme from the deep-living fish species. It was possible to conclude that the major adaptive changes to pressure tolerance must be located in the N-terminal region of the protein. The types of changes that are found and their spatial location within the protein structure are discussed. An analysis of the kinetic parameters of the enzymes suggests that there is clearly a trade off between Km and kcat values, which likely reflects the necessity of the deep-sea enzyme to operate at low temperatures. |
format |
Article in Journal/Newspaper |
author |
Brindley, AA Pickersgill, RW Partridge, JC Dunstan, DJ Hunt, DM Warren, MJ |
spellingShingle |
Brindley, AA Pickersgill, RW Partridge, JC Dunstan, DJ Hunt, DM Warren, MJ Enzyme Sequence and Its Relationship to Hyperbaric Stability of Artificial and Natural Fish Lactate Dehydrogenases |
author_facet |
Brindley, AA Pickersgill, RW Partridge, JC Dunstan, DJ Hunt, DM Warren, MJ |
author_sort |
Brindley, AA |
title |
Enzyme Sequence and Its Relationship to Hyperbaric Stability of Artificial and Natural Fish Lactate Dehydrogenases |
title_short |
Enzyme Sequence and Its Relationship to Hyperbaric Stability of Artificial and Natural Fish Lactate Dehydrogenases |
title_full |
Enzyme Sequence and Its Relationship to Hyperbaric Stability of Artificial and Natural Fish Lactate Dehydrogenases |
title_fullStr |
Enzyme Sequence and Its Relationship to Hyperbaric Stability of Artificial and Natural Fish Lactate Dehydrogenases |
title_full_unstemmed |
Enzyme Sequence and Its Relationship to Hyperbaric Stability of Artificial and Natural Fish Lactate Dehydrogenases |
title_sort |
enzyme sequence and its relationship to hyperbaric stability of artificial and natural fish lactate dehydrogenases |
publisher |
PUBLIC LIBRARY SCIENCE |
publishDate |
2008 |
url |
https://discovery.ucl.ac.uk/id/eprint/123018/1/123018.pdf https://discovery.ucl.ac.uk/id/eprint/123018/ |
genre |
Gadus morhua |
genre_facet |
Gadus morhua |
op_source |
PLOS ONE , 3 (4) , Article e2042. (2008) |
op_relation |
https://discovery.ucl.ac.uk/id/eprint/123018/1/123018.pdf https://discovery.ucl.ac.uk/id/eprint/123018/ |
op_rights |
open |
_version_ |
1786204600594857984 |