Enzymatic modification by point mutation and functional analysis of an omega-6 fatty acid desaturase from Arctic Chlamydomonas sp.
Arctic Chlamydomonas sp. is a dominant microalgal strain in cold or frozen freshwater in the Arctic region. The full-length open reading frame of the omega-6 fatty acid desaturase gene (AChFAD6) was obtained from the transcriptomic database of Arctic Chlamydomonas sp. from the KOPRI culture collecti...
Main Authors: | , , , , , |
---|---|
Format: | Text |
Language: | unknown |
Published: |
Taylor & Francis
2016
|
Subjects: | |
Online Access: | https://dx.doi.org/10.6084/m9.figshare.3384925 https://tandf.figshare.com/articles/journal_contribution/Enzymatic_modification_by_point_mutation_and_functional_analysis_of_an_omega-6_fatty_acid_desaturase_from_arctic_i_Chlamydomonas_i_sp_/3384925 |
id |
ftdatacite:10.6084/m9.figshare.3384925 |
---|---|
record_format |
openpolar |
spelling |
ftdatacite:10.6084/m9.figshare.3384925 2023-05-15T14:45:38+02:00 Enzymatic modification by point mutation and functional analysis of an omega-6 fatty acid desaturase from Arctic Chlamydomonas sp. Woongsic Jung Kim, Eun Jae Han, Se Jong Kang, Sung-Ho Han-Gu Choi Sanghee Kim 2016 https://dx.doi.org/10.6084/m9.figshare.3384925 https://tandf.figshare.com/articles/journal_contribution/Enzymatic_modification_by_point_mutation_and_functional_analysis_of_an_omega-6_fatty_acid_desaturase_from_arctic_i_Chlamydomonas_i_sp_/3384925 unknown Taylor & Francis https://dx.doi.org/10.1080/10826068.2016.1188311 Creative Commons Attribution 4.0 International https://creativecommons.org/licenses/by/4.0/legalcode cc-by-4.0 CC-BY Biochemistry Medicine Microbiology FOS Biological sciences Cell Biology Genetics 39999 Chemical Sciences not elsewhere classified FOS Chemical sciences 69999 Biological Sciences not elsewhere classified Developmental Biology Plant Biology Text article-journal Journal contribution ScholarlyArticle 2016 ftdatacite https://doi.org/10.6084/m9.figshare.3384925 https://doi.org/10.1080/10826068.2016.1188311 2021-11-05T12:55:41Z Arctic Chlamydomonas sp. is a dominant microalgal strain in cold or frozen freshwater in the Arctic region. The full-length open reading frame of the omega-6 fatty acid desaturase gene (AChFAD6) was obtained from the transcriptomic database of Arctic Chlamydomonas sp. from the KOPRI culture collection of polar micro-organisms. Amino acid sequence analysis indicated the presence of three conserved histidine-rich segments as unique characteristics of omega-6 fatty acid desaturases, and three transmembrane regions transported to plastidic membranes by chloroplast transit peptides in the N-terminal region. The AChFAD6 desaturase activity was examined by expressing wild-type and V254A mutant (Mut-AChFAD6) heterologous recombinant proteins. Quantitative gas chromatography indicated that the concentration of linoleic acids in AChFAD6 -transformed cells increased more than 3-fold [6.73 ± 0.13 mg g −1 dry cell weight (DCW)] compared with cells transformed with vector alone. In contrast, transformation with Mut- AChFAD6 increased the concentration of oleic acid to 9.23 ± 0.18 mg g −1 DCW, indicating a change in enzymatic activity to mimic that of stearoyl-CoA desaturase. These results demonstrate that AChFAD6 of Arctic Chlamydomonas sp. increases membrane fluidity by enhancing denaturation of C18 fatty acids and facilitates production of large quantities of linoleic fatty acids in prokaryotic expression systems. Text Arctic DataCite Metadata Store (German National Library of Science and Technology) Arctic |
institution |
Open Polar |
collection |
DataCite Metadata Store (German National Library of Science and Technology) |
op_collection_id |
ftdatacite |
language |
unknown |
topic |
Biochemistry Medicine Microbiology FOS Biological sciences Cell Biology Genetics 39999 Chemical Sciences not elsewhere classified FOS Chemical sciences 69999 Biological Sciences not elsewhere classified Developmental Biology Plant Biology |
spellingShingle |
Biochemistry Medicine Microbiology FOS Biological sciences Cell Biology Genetics 39999 Chemical Sciences not elsewhere classified FOS Chemical sciences 69999 Biological Sciences not elsewhere classified Developmental Biology Plant Biology Woongsic Jung Kim, Eun Jae Han, Se Jong Kang, Sung-Ho Han-Gu Choi Sanghee Kim Enzymatic modification by point mutation and functional analysis of an omega-6 fatty acid desaturase from Arctic Chlamydomonas sp. |
topic_facet |
Biochemistry Medicine Microbiology FOS Biological sciences Cell Biology Genetics 39999 Chemical Sciences not elsewhere classified FOS Chemical sciences 69999 Biological Sciences not elsewhere classified Developmental Biology Plant Biology |
description |
Arctic Chlamydomonas sp. is a dominant microalgal strain in cold or frozen freshwater in the Arctic region. The full-length open reading frame of the omega-6 fatty acid desaturase gene (AChFAD6) was obtained from the transcriptomic database of Arctic Chlamydomonas sp. from the KOPRI culture collection of polar micro-organisms. Amino acid sequence analysis indicated the presence of three conserved histidine-rich segments as unique characteristics of omega-6 fatty acid desaturases, and three transmembrane regions transported to plastidic membranes by chloroplast transit peptides in the N-terminal region. The AChFAD6 desaturase activity was examined by expressing wild-type and V254A mutant (Mut-AChFAD6) heterologous recombinant proteins. Quantitative gas chromatography indicated that the concentration of linoleic acids in AChFAD6 -transformed cells increased more than 3-fold [6.73 ± 0.13 mg g −1 dry cell weight (DCW)] compared with cells transformed with vector alone. In contrast, transformation with Mut- AChFAD6 increased the concentration of oleic acid to 9.23 ± 0.18 mg g −1 DCW, indicating a change in enzymatic activity to mimic that of stearoyl-CoA desaturase. These results demonstrate that AChFAD6 of Arctic Chlamydomonas sp. increases membrane fluidity by enhancing denaturation of C18 fatty acids and facilitates production of large quantities of linoleic fatty acids in prokaryotic expression systems. |
format |
Text |
author |
Woongsic Jung Kim, Eun Jae Han, Se Jong Kang, Sung-Ho Han-Gu Choi Sanghee Kim |
author_facet |
Woongsic Jung Kim, Eun Jae Han, Se Jong Kang, Sung-Ho Han-Gu Choi Sanghee Kim |
author_sort |
Woongsic Jung |
title |
Enzymatic modification by point mutation and functional analysis of an omega-6 fatty acid desaturase from Arctic Chlamydomonas sp. |
title_short |
Enzymatic modification by point mutation and functional analysis of an omega-6 fatty acid desaturase from Arctic Chlamydomonas sp. |
title_full |
Enzymatic modification by point mutation and functional analysis of an omega-6 fatty acid desaturase from Arctic Chlamydomonas sp. |
title_fullStr |
Enzymatic modification by point mutation and functional analysis of an omega-6 fatty acid desaturase from Arctic Chlamydomonas sp. |
title_full_unstemmed |
Enzymatic modification by point mutation and functional analysis of an omega-6 fatty acid desaturase from Arctic Chlamydomonas sp. |
title_sort |
enzymatic modification by point mutation and functional analysis of an omega-6 fatty acid desaturase from arctic chlamydomonas sp. |
publisher |
Taylor & Francis |
publishDate |
2016 |
url |
https://dx.doi.org/10.6084/m9.figshare.3384925 https://tandf.figshare.com/articles/journal_contribution/Enzymatic_modification_by_point_mutation_and_functional_analysis_of_an_omega-6_fatty_acid_desaturase_from_arctic_i_Chlamydomonas_i_sp_/3384925 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_relation |
https://dx.doi.org/10.1080/10826068.2016.1188311 |
op_rights |
Creative Commons Attribution 4.0 International https://creativecommons.org/licenses/by/4.0/legalcode cc-by-4.0 |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.6084/m9.figshare.3384925 https://doi.org/10.1080/10826068.2016.1188311 |
_version_ |
1766317014607986688 |