A Novel Cold-Adapted Leucine Dehydrogenase from Antarctic Sea-Ice Bacterium Pseudoalteromonas sp. ANT178
l-tert-leucine and its derivatives are useful as pharmaceutical active ingredients, in which leucine dehydrogenase (LeuDH) is the key enzyme in their enzymatic conversions. In the present study, a novel cold-adapted LeuDH, psleudh, was cloned from psychrotrophic bacteria Pseudoalteromonas sp. ANT178...
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ftmdpi:oai:mdpi.com:/1660-3397/16/10/359/ 2023-08-20T04:00:18+02:00 A Novel Cold-Adapted Leucine Dehydrogenase from Antarctic Sea-Ice Bacterium Pseudoalteromonas sp. ANT178 Yatong Wang Yanhua Hou Yifan Wang Lu Zheng Xianlei Xu Kang Pan Rongqi Li Quanfu Wang agris 2018-10-01 application/pdf https://doi.org/10.3390/md16100359 EN eng Multidisciplinary Digital Publishing Institute https://dx.doi.org/10.3390/md16100359 https://creativecommons.org/licenses/by/4.0/ Marine Drugs; Volume 16; Issue 10; Pages: 359 leucine dehydrogenase cold-adapted Antarctic bacterium sea-ice homology modeling Text 2018 ftmdpi https://doi.org/10.3390/md16100359 2023-07-31T21:45:29Z l-tert-leucine and its derivatives are useful as pharmaceutical active ingredients, in which leucine dehydrogenase (LeuDH) is the key enzyme in their enzymatic conversions. In the present study, a novel cold-adapted LeuDH, psleudh, was cloned from psychrotrophic bacteria Pseudoalteromonas sp. ANT178, which was isolated from Antarctic sea-ice. Bioinformatics analysis of the gene psleudh showed that the gene was 1209 bp in length and coded for a 42.6 kDa protein containing 402 amino acids. PsLeuDH had conserved Phe binding site and NAD+ binding site, and belonged to a member of the Glu/Leu/Phe/Val dehydrogenase family. Homology modeling analysis results suggested that PsLeuDH exhibited more glycine residues, reduced proline residues, and arginine residues, which might be responsible for its catalytic efficiency at low temperature. The recombinant PsLeuDH (rPsLeuDH) was purified a major band with the high specific activity of 275.13 U/mg using a Ni-NTA affinity chromatography. The optimum temperature and pH for rPsLeuDH activity were 30 °C and pH 9.0, respectively. Importantly, rPsLeuDH retained at least 40% of its maximum activity even at 0 °C. Moreover, the activity of rPsLeuDH was the highest in the presence of 2.0 M NaCl. Substrate specificity and kinetic studies of rPsLeuDH demonstrated that l-leucine was the most suitable substrate, and the catalytic activity at low temperatures was ensured by maintaining a high kcat value. The results of the current study would provide insight into Antarctic sea-ice bacterium LeuDH, and the unique properties of rPsLeuDH make it a promising candidate as a biocatalyst in medical and pharmaceutical industries. Text Antarc* Antarctic Sea ice MDPI Open Access Publishing Antarctic Marine Drugs 16 10 359 |
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Open Polar |
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MDPI Open Access Publishing |
op_collection_id |
ftmdpi |
language |
English |
topic |
leucine dehydrogenase cold-adapted Antarctic bacterium sea-ice homology modeling |
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leucine dehydrogenase cold-adapted Antarctic bacterium sea-ice homology modeling Yatong Wang Yanhua Hou Yifan Wang Lu Zheng Xianlei Xu Kang Pan Rongqi Li Quanfu Wang A Novel Cold-Adapted Leucine Dehydrogenase from Antarctic Sea-Ice Bacterium Pseudoalteromonas sp. ANT178 |
topic_facet |
leucine dehydrogenase cold-adapted Antarctic bacterium sea-ice homology modeling |
description |
l-tert-leucine and its derivatives are useful as pharmaceutical active ingredients, in which leucine dehydrogenase (LeuDH) is the key enzyme in their enzymatic conversions. In the present study, a novel cold-adapted LeuDH, psleudh, was cloned from psychrotrophic bacteria Pseudoalteromonas sp. ANT178, which was isolated from Antarctic sea-ice. Bioinformatics analysis of the gene psleudh showed that the gene was 1209 bp in length and coded for a 42.6 kDa protein containing 402 amino acids. PsLeuDH had conserved Phe binding site and NAD+ binding site, and belonged to a member of the Glu/Leu/Phe/Val dehydrogenase family. Homology modeling analysis results suggested that PsLeuDH exhibited more glycine residues, reduced proline residues, and arginine residues, which might be responsible for its catalytic efficiency at low temperature. The recombinant PsLeuDH (rPsLeuDH) was purified a major band with the high specific activity of 275.13 U/mg using a Ni-NTA affinity chromatography. The optimum temperature and pH for rPsLeuDH activity were 30 °C and pH 9.0, respectively. Importantly, rPsLeuDH retained at least 40% of its maximum activity even at 0 °C. Moreover, the activity of rPsLeuDH was the highest in the presence of 2.0 M NaCl. Substrate specificity and kinetic studies of rPsLeuDH demonstrated that l-leucine was the most suitable substrate, and the catalytic activity at low temperatures was ensured by maintaining a high kcat value. The results of the current study would provide insight into Antarctic sea-ice bacterium LeuDH, and the unique properties of rPsLeuDH make it a promising candidate as a biocatalyst in medical and pharmaceutical industries. |
format |
Text |
author |
Yatong Wang Yanhua Hou Yifan Wang Lu Zheng Xianlei Xu Kang Pan Rongqi Li Quanfu Wang |
author_facet |
Yatong Wang Yanhua Hou Yifan Wang Lu Zheng Xianlei Xu Kang Pan Rongqi Li Quanfu Wang |
author_sort |
Yatong Wang |
title |
A Novel Cold-Adapted Leucine Dehydrogenase from Antarctic Sea-Ice Bacterium Pseudoalteromonas sp. ANT178 |
title_short |
A Novel Cold-Adapted Leucine Dehydrogenase from Antarctic Sea-Ice Bacterium Pseudoalteromonas sp. ANT178 |
title_full |
A Novel Cold-Adapted Leucine Dehydrogenase from Antarctic Sea-Ice Bacterium Pseudoalteromonas sp. ANT178 |
title_fullStr |
A Novel Cold-Adapted Leucine Dehydrogenase from Antarctic Sea-Ice Bacterium Pseudoalteromonas sp. ANT178 |
title_full_unstemmed |
A Novel Cold-Adapted Leucine Dehydrogenase from Antarctic Sea-Ice Bacterium Pseudoalteromonas sp. ANT178 |
title_sort |
novel cold-adapted leucine dehydrogenase from antarctic sea-ice bacterium pseudoalteromonas sp. ant178 |
publisher |
Multidisciplinary Digital Publishing Institute |
publishDate |
2018 |
url |
https://doi.org/10.3390/md16100359 |
op_coverage |
agris |
geographic |
Antarctic |
geographic_facet |
Antarctic |
genre |
Antarc* Antarctic Sea ice |
genre_facet |
Antarc* Antarctic Sea ice |
op_source |
Marine Drugs; Volume 16; Issue 10; Pages: 359 |
op_relation |
https://dx.doi.org/10.3390/md16100359 |
op_rights |
https://creativecommons.org/licenses/by/4.0/ |
op_doi |
https://doi.org/10.3390/md16100359 |
container_title |
Marine Drugs |
container_volume |
16 |
container_issue |
10 |
container_start_page |
359 |
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1774717440957087744 |