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Molecular And 3D-Structural Characterization Of Fructose-1,6-Bisphosphate Aldolase Derived From Metroxylon Sagu

Fructose-1,6-bisphosphate aldolase (FBAld) is an enzyme that catalyzes the cleavage of D-fructose-1,6-phosphate (FBP) to D-glyceraldehyde-3-phosphate (G3P) and dihydroxyacetone phosphate (DHAP), and plays vital role in glycolysis and gluconeogenesis. However, molecular characterization and functiona...

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Main Authors: Hairul Azman, Roslan, Md. Anowar, Hossain, Jerry, anak Gerunsin
Format: E-Article
Language:English
Published: SCIELO 2017
Subjects:
Q Science (General)
Online Access:http://ir.unimas.my/id/eprint/17018/1/Molecular_And_3D-Structural_Characterization_Of_Fr%28Abstract%29.pdf
http://ir.unimas.my/id/eprint/17018/
http://dx.doi.org/10.1590/1678-4324-2017160108
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spelling my.unimas.ir.170182017-08-09T06:47:14Z http://ir.unimas.my/id/eprint/17018/ Molecular And 3D-Structural Characterization Of Fructose-1,6-Bisphosphate Aldolase Derived From Metroxylon Sagu Hairul Azman, Roslan Md. Anowar, Hossain Jerry, anak Gerunsin Q Science (General) Fructose-1,6-bisphosphate aldolase (FBAld) is an enzyme that catalyzes the cleavage of D-fructose-1,6-phosphate (FBP) to D-glyceraldehyde-3-phosphate (G3P) and dihydroxyacetone phosphate (DHAP), and plays vital role in glycolysis and gluconeogenesis. However, molecular characterization and functional roles of FBAld remain unknown in sago palm. Here we report a modified CTAB-RNA extraction method was developed for the isolation of good quality RNA (RIN>8) from sago leaves and the isolation of FBAld cDNA from sago palm. The isolated sago FBAld (msFBAld) cDNA has total length of 1288 bp with an open reading frame of 1020 bp and a predicted to encode for a protein of 340 amino acid resides. The predicted protein shared a high degree of homology with Class-I FBAld from other plants. Meanwhile, the msFBAld gene spanned 2322 bp and consisted of five exons. Conserved domain search identified fifteen catalytically important amino acids at the active site and phylogenetic tree revealed localization of msFBAld in the chloroplast. A molecular 3D-structure of msFBAld was generated by homology modeling and a Ramachandran plot with 86.7% of the residues in the core region, 13.4% in the allowed region with no residues in the disallowed region. The modeled structure is a homotetramer containing an /-TIM-barrel at the center. Superimposition of the model with Class-I aldolases identified a catalytic dyad, Lys209-Glu167, which could be involved in the Schiff's base formation and aldol condensation. Apart from that, overproduction of the recombinant msFBAld in Escherichia coli resulted in increased tolerance towards salinity. SCIELO 2017 E-Article PeerReviewed text en http://ir.unimas.my/id/eprint/17018/1/Molecular_And_3D-Structural_Characterization_Of_Fr%28Abstract%29.pdf Hairul Azman, Roslan and Md. Anowar, Hossain and Jerry, anak Gerunsin (2017) Molecular And 3D-Structural Characterization Of Fructose-1,6-Bisphosphate Aldolase Derived From Metroxylon Sagu. Brazilian Archives of Biology and Technology, 60. ISSN 1516-8913 http://dx.doi.org/10.1590/1678-4324-2017160108 10.1590/1678-4324-2017160108
institution Universiti Malaysia Sarawak
building Centre for Academic Information Services (CAIS)
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaysia Sarawak
content_source UNIMAS Institutional Repository
url_provider http://ir.unimas.my/
language English
topic Q Science (General)
spellingShingle Q Science (General)
Hairul Azman, Roslan
Md. Anowar, Hossain
Jerry, anak Gerunsin
Molecular And 3D-Structural Characterization Of Fructose-1,6-Bisphosphate Aldolase Derived From Metroxylon Sagu
description Fructose-1,6-bisphosphate aldolase (FBAld) is an enzyme that catalyzes the cleavage of D-fructose-1,6-phosphate (FBP) to D-glyceraldehyde-3-phosphate (G3P) and dihydroxyacetone phosphate (DHAP), and plays vital role in glycolysis and gluconeogenesis. However, molecular characterization and functional roles of FBAld remain unknown in sago palm. Here we report a modified CTAB-RNA extraction method was developed for the isolation of good quality RNA (RIN>8) from sago leaves and the isolation of FBAld cDNA from sago palm. The isolated sago FBAld (msFBAld) cDNA has total length of 1288 bp with an open reading frame of 1020 bp and a predicted to encode for a protein of 340 amino acid resides. The predicted protein shared a high degree of homology with Class-I FBAld from other plants. Meanwhile, the msFBAld gene spanned 2322 bp and consisted of five exons. Conserved domain search identified fifteen catalytically important amino acids at the active site and phylogenetic tree revealed localization of msFBAld in the chloroplast. A molecular 3D-structure of msFBAld was generated by homology modeling and a Ramachandran plot with 86.7% of the residues in the core region, 13.4% in the allowed region with no residues in the disallowed region. The modeled structure is a homotetramer containing an /-TIM-barrel at the center. Superimposition of the model with Class-I aldolases identified a catalytic dyad, Lys209-Glu167, which could be involved in the Schiff's base formation and aldol condensation. Apart from that, overproduction of the recombinant msFBAld in Escherichia coli resulted in increased tolerance towards salinity.
format E-Article
author Hairul Azman, Roslan
Md. Anowar, Hossain
Jerry, anak Gerunsin
author_facet Hairul Azman, Roslan
Md. Anowar, Hossain
Jerry, anak Gerunsin
author_sort Hairul Azman, Roslan
title Molecular And 3D-Structural Characterization Of Fructose-1,6-Bisphosphate Aldolase Derived From Metroxylon Sagu
title_short Molecular And 3D-Structural Characterization Of Fructose-1,6-Bisphosphate Aldolase Derived From Metroxylon Sagu
title_full Molecular And 3D-Structural Characterization Of Fructose-1,6-Bisphosphate Aldolase Derived From Metroxylon Sagu
title_fullStr Molecular And 3D-Structural Characterization Of Fructose-1,6-Bisphosphate Aldolase Derived From Metroxylon Sagu
title_full_unstemmed Molecular And 3D-Structural Characterization Of Fructose-1,6-Bisphosphate Aldolase Derived From Metroxylon Sagu
title_sort molecular and 3d-structural characterization of fructose-1,6-bisphosphate aldolase derived from metroxylon sagu
publisher SCIELO
publishDate 2017
url http://ir.unimas.my/id/eprint/17018/1/Molecular_And_3D-Structural_Characterization_Of_Fr%28Abstract%29.pdf
http://ir.unimas.my/id/eprint/17018/
http://dx.doi.org/10.1590/1678-4324-2017160108
_version_ 1644512508159983616
score 13.235362

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