Cold-Adapted Alanine Dehydrogenases from Two Antarctic Bacterial Strains: Gene Cloning, Protein Characterization, and Comparison with Mesophilic and Thermophilic Counterparts
The genes encoding NAD+-dependent alanine dehydrogenases (AlaDHs) (EC 1.4.1.1) from the Antarctic bacterial organisms Shewanella sp. strain Ac10 (SheAlaDH) and Carnobacterium sp. strain St2 (CarAlaDH) were cloned and expressed in Escherichia coli. Of all of the AlaDHs that have been sequenced, SheAl...
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ftpubmed:oai:pubmedcentral.nih.gov:99735 2023-05-15T14:03:18+02:00 Cold-Adapted Alanine Dehydrogenases from Two Antarctic Bacterial Strains: Gene Cloning, Protein Characterization, and Comparison with Mesophilic and Thermophilic Counterparts Galkin, Andrey Kulakova, Ljudmila Ashida, Hiroyuki Sawa, Yoshihiro Esaki, Nobuyoshi 1999-09 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC99735 http://www.ncbi.nlm.nih.gov/pubmed/10473410 en eng American Society for Microbiology http://www.ncbi.nlm.nih.gov/pmc/articles/PMC99735 http://www.ncbi.nlm.nih.gov/pubmed/10473410 Copyright © 1999, American Society for Microbiology Enzymology and Protein Engineering Text 1999 ftpubmed 2013-08-29T09:45:58Z The genes encoding NAD+-dependent alanine dehydrogenases (AlaDHs) (EC 1.4.1.1) from the Antarctic bacterial organisms Shewanella sp. strain Ac10 (SheAlaDH) and Carnobacterium sp. strain St2 (CarAlaDH) were cloned and expressed in Escherichia coli. Of all of the AlaDHs that have been sequenced, SheAlaDH exhibited the highest level of sequence similarity to the AlaDH from the gram-negative bacterium Vibrio proteolyticus (VprAlaDH). CarAlaDH was most similar to AlaDHs from mesophilic and thermophilic Bacillus strains. SheAlaDH and CarAlaDH had features typical of cold-adapted enzymes; both the optimal temperature for catalytic activity and the temperature limit for retaining thermostability were lower than the values obtained for the mesophilic counterparts. The kcat/Km value for the SheAlaDH reaction was about three times higher than the kcat/Km value for VprAlaDH, but it was much lower than the kcat/Km value for the AlaDH from Bacillus subtilis. Homology-based structural models of various AlaDHs, including the two psychrotrophic AlaDHs, were constructed. The thermal instability of SheAlaDH and CarAlaDH may result from relatively low numbers of salt bridges in these proteins. Text Antarc* Antarctic PubMed Central (PMC) Antarctic The Antarctic |
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Enzymology and Protein Engineering |
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Enzymology and Protein Engineering Galkin, Andrey Kulakova, Ljudmila Ashida, Hiroyuki Sawa, Yoshihiro Esaki, Nobuyoshi Cold-Adapted Alanine Dehydrogenases from Two Antarctic Bacterial Strains: Gene Cloning, Protein Characterization, and Comparison with Mesophilic and Thermophilic Counterparts |
topic_facet |
Enzymology and Protein Engineering |
description |
The genes encoding NAD+-dependent alanine dehydrogenases (AlaDHs) (EC 1.4.1.1) from the Antarctic bacterial organisms Shewanella sp. strain Ac10 (SheAlaDH) and Carnobacterium sp. strain St2 (CarAlaDH) were cloned and expressed in Escherichia coli. Of all of the AlaDHs that have been sequenced, SheAlaDH exhibited the highest level of sequence similarity to the AlaDH from the gram-negative bacterium Vibrio proteolyticus (VprAlaDH). CarAlaDH was most similar to AlaDHs from mesophilic and thermophilic Bacillus strains. SheAlaDH and CarAlaDH had features typical of cold-adapted enzymes; both the optimal temperature for catalytic activity and the temperature limit for retaining thermostability were lower than the values obtained for the mesophilic counterparts. The kcat/Km value for the SheAlaDH reaction was about three times higher than the kcat/Km value for VprAlaDH, but it was much lower than the kcat/Km value for the AlaDH from Bacillus subtilis. Homology-based structural models of various AlaDHs, including the two psychrotrophic AlaDHs, were constructed. The thermal instability of SheAlaDH and CarAlaDH may result from relatively low numbers of salt bridges in these proteins. |
format |
Text |
author |
Galkin, Andrey Kulakova, Ljudmila Ashida, Hiroyuki Sawa, Yoshihiro Esaki, Nobuyoshi |
author_facet |
Galkin, Andrey Kulakova, Ljudmila Ashida, Hiroyuki Sawa, Yoshihiro Esaki, Nobuyoshi |
author_sort |
Galkin, Andrey |
title |
Cold-Adapted Alanine Dehydrogenases from Two Antarctic Bacterial Strains: Gene Cloning, Protein Characterization, and Comparison with Mesophilic and Thermophilic Counterparts |
title_short |
Cold-Adapted Alanine Dehydrogenases from Two Antarctic Bacterial Strains: Gene Cloning, Protein Characterization, and Comparison with Mesophilic and Thermophilic Counterparts |
title_full |
Cold-Adapted Alanine Dehydrogenases from Two Antarctic Bacterial Strains: Gene Cloning, Protein Characterization, and Comparison with Mesophilic and Thermophilic Counterparts |
title_fullStr |
Cold-Adapted Alanine Dehydrogenases from Two Antarctic Bacterial Strains: Gene Cloning, Protein Characterization, and Comparison with Mesophilic and Thermophilic Counterparts |
title_full_unstemmed |
Cold-Adapted Alanine Dehydrogenases from Two Antarctic Bacterial Strains: Gene Cloning, Protein Characterization, and Comparison with Mesophilic and Thermophilic Counterparts |
title_sort |
cold-adapted alanine dehydrogenases from two antarctic bacterial strains: gene cloning, protein characterization, and comparison with mesophilic and thermophilic counterparts |
publisher |
American Society for Microbiology |
publishDate |
1999 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC99735 http://www.ncbi.nlm.nih.gov/pubmed/10473410 |
geographic |
Antarctic The Antarctic |
geographic_facet |
Antarctic The Antarctic |
genre |
Antarc* Antarctic |
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Antarc* Antarctic |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC99735 http://www.ncbi.nlm.nih.gov/pubmed/10473410 |
op_rights |
Copyright © 1999, American Society for Microbiology |
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