Cold-Adapted Alanine Dehydrogenases from Two Antarctic Bacterial Strains: Gene Cloning, Protein Characterization, and Comparison with Mesophilic and Thermophilic Counterparts
ABSTRACT 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 seq...
Published in: | Applied and Environmental Microbiology |
---|---|
Main Authors: | , , , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
American Society for Microbiology
1999
|
Subjects: | |
Online Access: | http://dx.doi.org/10.1128/aem.65.9.4014-4020.1999 https://journals.asm.org/doi/pdf/10.1128/AEM.65.9.4014-4020.1999 |
id |
crasmicro:10.1128/aem.65.9.4014-4020.1999 |
---|---|
record_format |
openpolar |
spelling |
crasmicro:10.1128/aem.65.9.4014-4020.1999 2023-11-05T03:36:31+01:00 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 http://dx.doi.org/10.1128/aem.65.9.4014-4020.1999 https://journals.asm.org/doi/pdf/10.1128/AEM.65.9.4014-4020.1999 en eng American Society for Microbiology https://journals.asm.org/non-commercial-tdm-license Applied and Environmental Microbiology volume 65, issue 9, page 4014-4020 ISSN 0099-2240 1098-5336 Ecology Applied Microbiology and Biotechnology Food Science Biotechnology journal-article 1999 crasmicro https://doi.org/10.1128/aem.65.9.4014-4020.1999 2023-10-09T16:02:23Z ABSTRACT 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 k cat / K m value for the SheAlaDH reaction was about three times higher than the k cat / K m value for VprAlaDH, but it was much lower than the k cat / K m 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. Article in Journal/Newspaper Antarc* Antarctic ASM Journals (American Society for Microbiology - via Crossref) Applied and Environmental Microbiology 65 9 4014 4020 |
institution |
Open Polar |
collection |
ASM Journals (American Society for Microbiology - via Crossref) |
op_collection_id |
crasmicro |
language |
English |
topic |
Ecology Applied Microbiology and Biotechnology Food Science Biotechnology |
spellingShingle |
Ecology Applied Microbiology and Biotechnology Food Science Biotechnology 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 |
Ecology Applied Microbiology and Biotechnology Food Science Biotechnology |
description |
ABSTRACT 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 k cat / K m value for the SheAlaDH reaction was about three times higher than the k cat / K m value for VprAlaDH, but it was much lower than the k cat / K m 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 |
Article in Journal/Newspaper |
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://dx.doi.org/10.1128/aem.65.9.4014-4020.1999 https://journals.asm.org/doi/pdf/10.1128/AEM.65.9.4014-4020.1999 |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
Applied and Environmental Microbiology volume 65, issue 9, page 4014-4020 ISSN 0099-2240 1098-5336 |
op_rights |
https://journals.asm.org/non-commercial-tdm-license |
op_doi |
https://doi.org/10.1128/aem.65.9.4014-4020.1999 |
container_title |
Applied and Environmental Microbiology |
container_volume |
65 |
container_issue |
9 |
container_start_page |
4014 |
op_container_end_page |
4020 |
_version_ |
1781691472647553024 |