Characterization and molecular cloning of secreted α-amylase with dominant activity from mon thong durian (Durio zibethinus murr. cv. mon thong)
Autor(a) principal: | |
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Data de Publicação: | 2021 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Revista brasileira de fruticultura (Online) |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-29452021000300801 |
Resumo: | Abstract The secreted α-amylase with dominant activity was purified from the crude extract of Mon Thong durian by steps of ammonium sulphate precipitation and the affinity column chromatography. The purified α-amylase (DzAmy1) had a molecular mass of approximately 44 kDa. Its optimum pH and temperature for activity were 7.0 and 50°C, respectively. The enzyme was stable from pH 6 to 10 and from 30 to 60°C. Many metal ions did not affect amylase activity. The gene cloning of DzAmy1 was carried out and it was confirmed that DzAmy1 gene consisted of 1,254 bp open reading frame, which encoded 23 amino acids of the signal peptide and 395 amino acids of mature protein with a calculated molecular mass of 43.7 kDa. The isoelectric point of the enzyme was 5.78. DzAmy1 was shown to belong to sub-family one of the plant α-amylases based on phylogenetic tree analysis. Structural characterization by homology modelling suggested that it consisted of 3 domains with a catalytic triad in domain A. Recombinant DzAmy1 (rDzAmy1) was successfully expressed in Escherichia coli and had hydrolysis activity for starch and ethylidene-pNP-G7, which was clearly confirmed the authenticity of DzAmy1 as a functional α-amylase. |
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Characterization and molecular cloning of secreted α-amylase with dominant activity from mon thong durian (Durio zibethinus murr. cv. mon thong)Durio zibethinus Murr. cv. Mon Thongsecreted α-amylasecharacterizationEscherichia coliAbstract The secreted α-amylase with dominant activity was purified from the crude extract of Mon Thong durian by steps of ammonium sulphate precipitation and the affinity column chromatography. The purified α-amylase (DzAmy1) had a molecular mass of approximately 44 kDa. Its optimum pH and temperature for activity were 7.0 and 50°C, respectively. The enzyme was stable from pH 6 to 10 and from 30 to 60°C. Many metal ions did not affect amylase activity. The gene cloning of DzAmy1 was carried out and it was confirmed that DzAmy1 gene consisted of 1,254 bp open reading frame, which encoded 23 amino acids of the signal peptide and 395 amino acids of mature protein with a calculated molecular mass of 43.7 kDa. The isoelectric point of the enzyme was 5.78. DzAmy1 was shown to belong to sub-family one of the plant α-amylases based on phylogenetic tree analysis. Structural characterization by homology modelling suggested that it consisted of 3 domains with a catalytic triad in domain A. Recombinant DzAmy1 (rDzAmy1) was successfully expressed in Escherichia coli and had hydrolysis activity for starch and ethylidene-pNP-G7, which was clearly confirmed the authenticity of DzAmy1 as a functional α-amylase.Sociedade Brasileira de Fruticultura2021-01-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-29452021000300801Revista Brasileira de Fruticultura v.43 n.3 2021reponame:Revista brasileira de fruticultura (Online)instname:Sociedade Brasileira de Fruticultura (SBF)instacron:SBFRU10.1590/0100-29452021231info:eu-repo/semantics/openAccessPosoongnoen,SaijaiUbonbal,RaksmontKlaynongsruang,SompongDaduang,JureerutRoytrakul,SittirukDaduang,Sakdaeng2021-06-16T00:00:00Zoai:scielo:S0100-29452021000300801Revistahttp://www.scielo.br/rbfhttps://old.scielo.br/oai/scielo-oai.phprbf@fcav.unesp.br||http://rbf.org.br/1806-99670100-2945opendoar:2021-06-16T00:00Revista brasileira de fruticultura (Online) - Sociedade Brasileira de Fruticultura (SBF)false |
dc.title.none.fl_str_mv |
Characterization and molecular cloning of secreted α-amylase with dominant activity from mon thong durian (Durio zibethinus murr. cv. mon thong) |
title |
Characterization and molecular cloning of secreted α-amylase with dominant activity from mon thong durian (Durio zibethinus murr. cv. mon thong) |
spellingShingle |
Characterization and molecular cloning of secreted α-amylase with dominant activity from mon thong durian (Durio zibethinus murr. cv. mon thong) Posoongnoen,Saijai Durio zibethinus Murr. cv. Mon Thong secreted α-amylase characterization Escherichia coli |
title_short |
Characterization and molecular cloning of secreted α-amylase with dominant activity from mon thong durian (Durio zibethinus murr. cv. mon thong) |
title_full |
Characterization and molecular cloning of secreted α-amylase with dominant activity from mon thong durian (Durio zibethinus murr. cv. mon thong) |
title_fullStr |
Characterization and molecular cloning of secreted α-amylase with dominant activity from mon thong durian (Durio zibethinus murr. cv. mon thong) |
title_full_unstemmed |
Characterization and molecular cloning of secreted α-amylase with dominant activity from mon thong durian (Durio zibethinus murr. cv. mon thong) |
title_sort |
Characterization and molecular cloning of secreted α-amylase with dominant activity from mon thong durian (Durio zibethinus murr. cv. mon thong) |
author |
Posoongnoen,Saijai |
author_facet |
Posoongnoen,Saijai Ubonbal,Raksmont Klaynongsruang,Sompong Daduang,Jureerut Roytrakul,Sittiruk Daduang,Sakda |
author_role |
author |
author2 |
Ubonbal,Raksmont Klaynongsruang,Sompong Daduang,Jureerut Roytrakul,Sittiruk Daduang,Sakda |
author2_role |
author author author author author |
dc.contributor.author.fl_str_mv |
Posoongnoen,Saijai Ubonbal,Raksmont Klaynongsruang,Sompong Daduang,Jureerut Roytrakul,Sittiruk Daduang,Sakda |
dc.subject.por.fl_str_mv |
Durio zibethinus Murr. cv. Mon Thong secreted α-amylase characterization Escherichia coli |
topic |
Durio zibethinus Murr. cv. Mon Thong secreted α-amylase characterization Escherichia coli |
description |
Abstract The secreted α-amylase with dominant activity was purified from the crude extract of Mon Thong durian by steps of ammonium sulphate precipitation and the affinity column chromatography. The purified α-amylase (DzAmy1) had a molecular mass of approximately 44 kDa. Its optimum pH and temperature for activity were 7.0 and 50°C, respectively. The enzyme was stable from pH 6 to 10 and from 30 to 60°C. Many metal ions did not affect amylase activity. The gene cloning of DzAmy1 was carried out and it was confirmed that DzAmy1 gene consisted of 1,254 bp open reading frame, which encoded 23 amino acids of the signal peptide and 395 amino acids of mature protein with a calculated molecular mass of 43.7 kDa. The isoelectric point of the enzyme was 5.78. DzAmy1 was shown to belong to sub-family one of the plant α-amylases based on phylogenetic tree analysis. Structural characterization by homology modelling suggested that it consisted of 3 domains with a catalytic triad in domain A. Recombinant DzAmy1 (rDzAmy1) was successfully expressed in Escherichia coli and had hydrolysis activity for starch and ethylidene-pNP-G7, which was clearly confirmed the authenticity of DzAmy1 as a functional α-amylase. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-01-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-29452021000300801 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-29452021000300801 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/0100-29452021231 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Sociedade Brasileira de Fruticultura |
publisher.none.fl_str_mv |
Sociedade Brasileira de Fruticultura |
dc.source.none.fl_str_mv |
Revista Brasileira de Fruticultura v.43 n.3 2021 reponame:Revista brasileira de fruticultura (Online) instname:Sociedade Brasileira de Fruticultura (SBF) instacron:SBFRU |
instname_str |
Sociedade Brasileira de Fruticultura (SBF) |
instacron_str |
SBFRU |
institution |
SBFRU |
reponame_str |
Revista brasileira de fruticultura (Online) |
collection |
Revista brasileira de fruticultura (Online) |
repository.name.fl_str_mv |
Revista brasileira de fruticultura (Online) - Sociedade Brasileira de Fruticultura (SBF) |
repository.mail.fl_str_mv |
rbf@fcav.unesp.br||http://rbf.org.br/ |
_version_ |
1752122496515047424 |