Enzymatic characterization of two acetyl-CoA synthetase genes from Populus trichocarpa.
Identifieur interne : 001891 ( Main/Exploration ); précédent : 001890; suivant : 001892Enzymatic characterization of two acetyl-CoA synthetase genes from Populus trichocarpa.
Auteurs : Shan Cao [République populaire de Chine] ; Hui Li [République populaire de Chine] ; Xiaoyun Yao [République populaire de Chine] ; Lihong Li [République populaire de Chine] ; Luyao Jiang [République populaire de Chine] ; Qiang Zhang [République populaire de Chine] ; Jiaxue Zhang [République populaire de Chine] ; Di Liu [République populaire de Chine] ; Hai Lu [République populaire de Chine]Source :
- SpringerPlus [ 2193-1801 ] ; 2016.
Abstract
The acetyl-CoA synthetase (ACS) family is a subfamily of adenylate-forming enzymes, which has a close evolutionary relationship with the 4-coumarate:CoA ligase (4CL) family. In this study, two ACS genes were cloned from Populus trichocarpa and were named PtrACS1 and PtrACS2. Bioinformatics characterization of PtrACS1 and PtrACS2 showed that they contained the key ACS residues and a putative peroxisome targeting sequence 1 (PTS1) at the end of the C-terminal sequence. Real-time PCR results showed that PtrACS1 and PtrACS2 were expressed in the phloem, xylem, leaves, and roots of one-year-old P. trichocarpa, but were expressed primarily in the leaves. The ACS enzyme activity was higher in leaves than other tissues in P. trichocarpa. Two overexpressed recombinant proteins showed no catalytic activity toward the substrates of 4CL, but did have notable catalytic activity toward sodium acetate and substrates of ACS. The relative activities of PtrACS1 and PtrACS2 were 194.16 ± 11.23 and 422.25 ± 21.69 μM min(-1) mg(-1), respectively. The K m and V max of PtrACS1 were 0.25 mM and 698.85 μM min(-1) mg(-1), while those for PtrACS2 were 0.72 mM and 245.96 μM min(-1) mg(-1), respectively. Our results revealed that both proteins belong to the ACS family, and provide a theoretical foundation for the identification and functional analysis of members of the adenylate-forming enzyme superfamily.
DOI: 10.1186/s40064-016-2532-7
PubMed: 27390658
PubMed Central: PMC4916118
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sort="Cao, Shan" uniqKey="Cao S" first="Shan" last="Cao">Shan Cao</name><affiliation wicri:level="3"><nlm:affiliation>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation><affiliation wicri:level="3"><nlm:affiliation>National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>National Engineering Laboratory for Tree Breeding, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Li, Hui" sort="Li, Hui" uniqKey="Li H" first="Hui" last="Li">Hui Li</name><affiliation wicri:level="3"><nlm:affiliation>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation><affiliation wicri:level="3"><nlm:affiliation>National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>National Engineering Laboratory for Tree Breeding, 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People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>National Engineering Laboratory for Tree Breeding, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Zhang, Qiang" sort="Zhang, Qiang" uniqKey="Zhang Q" first="Qiang" last="Zhang">Qiang Zhang</name><affiliation wicri:level="3"><nlm:affiliation>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation><affiliation wicri:level="3"><nlm:affiliation>National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>National Engineering Laboratory for Tree Breeding, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Zhang, Jiaxue" sort="Zhang, Jiaxue" uniqKey="Zhang J" first="Jiaxue" last="Zhang">Jiaxue Zhang</name><affiliation wicri:level="3"><nlm:affiliation>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation><affiliation wicri:level="3"><nlm:affiliation>National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>National Engineering Laboratory for Tree Breeding, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Liu, Di" sort="Liu, Di" uniqKey="Liu D" first="Di" last="Liu">Di Liu</name><affiliation wicri:level="3"><nlm:affiliation>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author><author><name sortKey="Lu, Hai" sort="Lu, Hai" uniqKey="Lu H" first="Hai" last="Lu">Hai Lu</name><affiliation wicri:level="3"><nlm:affiliation>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation><affiliation wicri:level="3"><nlm:affiliation>National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>National Engineering Laboratory for Tree Breeding, Beijing</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName></affiliation></author></analytic><series><title level="j">SpringerPlus</title><idno type="ISSN">2193-1801</idno><imprint><date when="2016" type="published">2016</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The acetyl-CoA synthetase (ACS) family is a subfamily of adenylate-forming enzymes, which has a close evolutionary relationship with the 4-coumarate:CoA ligase (4CL) family. In this study, two ACS genes were cloned from Populus trichocarpa and were named PtrACS1 and PtrACS2. Bioinformatics characterization of PtrACS1 and PtrACS2 showed that they contained the key ACS residues and a putative peroxisome targeting sequence 1 (PTS1) at the end of the C-terminal sequence. Real-time PCR results showed that PtrACS1 and PtrACS2 were expressed in the phloem, xylem, leaves, and roots of one-year-old P. trichocarpa, but were expressed primarily in the leaves. The ACS enzyme activity was higher in leaves than other tissues in P. trichocarpa. Two overexpressed recombinant proteins showed no catalytic activity toward the substrates of 4CL, but did have notable catalytic activity toward sodium acetate and substrates of ACS. The relative activities of PtrACS1 and PtrACS2 were 194.16 ± 11.23 and 422.25 ± 21.69 μM min(-1) mg(-1), respectively. The K m and V max of PtrACS1 were 0.25 mM and 698.85 μM min(-1) mg(-1), while those for PtrACS2 were 0.72 mM and 245.96 μM min(-1) mg(-1), respectively. Our results revealed that both proteins belong to the ACS family, and provide a theoretical foundation for the identification and functional analysis of members of the adenylate-forming enzyme superfamily. </div></front></TEI><pubmed><MedlineCitation Status="PubMed-not-MEDLINE" Owner="NLM"><PMID Version="1">27390658</PMID><DateCompleted><Year>2016</Year><Month>07</Month><Day>08</Day></DateCompleted><DateRevised><Year>2020</Year><Month>10</Month><Day>01</Day></DateRevised><Article PubModel="Electronic-eCollection"><Journal><ISSN IssnType="Print">2193-1801</ISSN><JournalIssue CitedMedium="Print"><Volume>5</Volume><Issue>1</Issue><PubDate><Year>2016</Year></PubDate></JournalIssue><Title>SpringerPlus</Title><ISOAbbreviation>Springerplus</ISOAbbreviation></Journal><ArticleTitle>Enzymatic characterization of two acetyl-CoA synthetase genes from Populus trichocarpa.</ArticleTitle><Pagination><MedlinePgn>818</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1186/s40064-016-2532-7</ELocationID><Abstract><AbstractText>The acetyl-CoA synthetase (ACS) family is a subfamily of adenylate-forming enzymes, which has a close evolutionary relationship with the 4-coumarate:CoA ligase (4CL) family. In this study, two ACS genes were cloned from Populus trichocarpa and were named PtrACS1 and PtrACS2. Bioinformatics characterization of PtrACS1 and PtrACS2 showed that they contained the key ACS residues and a putative peroxisome targeting sequence 1 (PTS1) at the end of the C-terminal sequence. Real-time PCR results showed that PtrACS1 and PtrACS2 were expressed in the phloem, xylem, leaves, and roots of one-year-old P. trichocarpa, but were expressed primarily in the leaves. The ACS enzyme activity was higher in leaves than other tissues in P. trichocarpa. Two overexpressed recombinant proteins showed no catalytic activity toward the substrates of 4CL, but did have notable catalytic activity toward sodium acetate and substrates of ACS. The relative activities of PtrACS1 and PtrACS2 were 194.16 ± 11.23 and 422.25 ± 21.69 μM min(-1) mg(-1), respectively. The K m and V max of PtrACS1 were 0.25 mM and 698.85 μM min(-1) mg(-1), while those for PtrACS2 were 0.72 mM and 245.96 μM min(-1) mg(-1), respectively. Our results revealed that both proteins belong to the ACS family, and provide a theoretical foundation for the identification and functional analysis of members of the adenylate-forming enzyme superfamily. </AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y" EqualContrib="Y"><LastName>Cao</LastName><ForeName>Shan</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y" EqualContrib="Y"><LastName>Li</LastName><ForeName>Hui</ForeName><Initials>H</Initials><AffiliationInfo><Affiliation>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y" EqualContrib="Y"><LastName>Yao</LastName><ForeName>Xiaoyun</ForeName><Initials>X</Initials><AffiliationInfo><Affiliation>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y" EqualContrib="Y"><LastName>Li</LastName><ForeName>Lihong</ForeName><Initials>L</Initials><AffiliationInfo><Affiliation>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Jiang</LastName><ForeName>Luyao</ForeName><Initials>L</Initials><AffiliationInfo><Affiliation>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Qiang</ForeName><Initials>Q</Initials><AffiliationInfo><Affiliation>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Jiaxue</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Liu</LastName><ForeName>Di</ForeName><Initials>D</Initials><AffiliationInfo><Affiliation>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Lu</LastName><ForeName>Hai</ForeName><Initials>H</Initials><AffiliationInfo><Affiliation>College of Life Sciences and Biotechnology, Beijing Forestry University, No. 35 Qinghua East Road, Haidian District, Beijing, 100083 People's Republic of China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>National Engineering Laboratory for Tree Breeding, Beijing, 100083 People's Republic of China.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2016</Year><Month>06</Month><Day>21</Day></ArticleDate></Article><MedlineJournalInfo><Country>Switzerland</Country><MedlineTA>Springerplus</MedlineTA><NlmUniqueID>101597967</NlmUniqueID><ISSNLinking>2193-1801</ISSNLinking></MedlineJournalInfo><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Acetyl-CoA synthetase (ACS)</Keyword><Keyword MajorTopicYN="N">Enzyme activity</Keyword><Keyword MajorTopicYN="N">Populus trichocarpa</Keyword><Keyword MajorTopicYN="N">Prokaryotic expression</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2016</Year><Month>01</Month><Day>12</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2016</Year><Month>06</Month><Day>07</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2016</Year><Month>7</Month><Day>9</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2016</Year><Month>7</Month><Day>9</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2016</Year><Month>7</Month><Day>9</Day><Hour>6</Hour><Minute>1</Minute></PubMedPubDate></History><PublicationStatus>epublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">27390658</ArticleId><ArticleId 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first="Luyao" last="Jiang">Luyao Jiang</name><name sortKey="Li, Hui" sort="Li, Hui" uniqKey="Li H" first="Hui" last="Li">Hui Li</name><name sortKey="Li, Hui" sort="Li, Hui" uniqKey="Li H" first="Hui" last="Li">Hui Li</name><name sortKey="Li, Lihong" sort="Li, Lihong" uniqKey="Li L" first="Lihong" last="Li">Lihong Li</name><name sortKey="Li, Lihong" sort="Li, Lihong" uniqKey="Li L" first="Lihong" last="Li">Lihong Li</name><name sortKey="Liu, Di" sort="Liu, Di" uniqKey="Liu D" first="Di" last="Liu">Di Liu</name><name sortKey="Lu, Hai" sort="Lu, Hai" uniqKey="Lu H" first="Hai" last="Lu">Hai Lu</name><name sortKey="Lu, Hai" sort="Lu, Hai" uniqKey="Lu H" first="Hai" last="Lu">Hai Lu</name><name sortKey="Yao, Xiaoyun" sort="Yao, Xiaoyun" uniqKey="Yao X" first="Xiaoyun" last="Yao">Xiaoyun Yao</name><name sortKey="Yao, Xiaoyun" sort="Yao, Xiaoyun" uniqKey="Yao X" first="Xiaoyun" last="Yao">Xiaoyun Yao</name><name sortKey="Zhang, Jiaxue" sort="Zhang, Jiaxue" uniqKey="Zhang J" first="Jiaxue" last="Zhang">Jiaxue Zhang</name><name sortKey="Zhang, Jiaxue" sort="Zhang, Jiaxue" uniqKey="Zhang J" first="Jiaxue" last="Zhang">Jiaxue Zhang</name><name sortKey="Zhang, Qiang" sort="Zhang, Qiang" uniqKey="Zhang Q" first="Qiang" last="Zhang">Qiang Zhang</name><name sortKey="Zhang, Qiang" sort="Zhang, Qiang" uniqKey="Zhang Q" first="Qiang" last="Zhang">Qiang Zhang</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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