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Gene and structural characterisation of pea cDNA library for pullulanase

Pullulanases belong to a discrete class of debranching enzymes that are capable of hydrolysing the α-1,6-branches in starch. This work described the screening, identification and characterization of pullulanase gene from pea cDNA library. The screening process used degenerate primers based on the al...

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Main Authors: Hasnain, Hussain, Zainab, Ngaini
Format: Proceeding
Language:English
Published: 2011
Subjects:
Q Science (General)
QR Microbiology
Online Access:http://ir.unimas.my/id/eprint/15580/1/Gene%20and%20structural.pdf
http://ir.unimas.my/id/eprint/15580/
https://www.researchgate.net/publication/291341950_Gene_and_structural_characterisation_of_pea_cDNA_library_for_pullulanase
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spelling my.unimas.ir.155802022-01-04T06:52:31Z http://ir.unimas.my/id/eprint/15580/ Gene and structural characterisation of pea cDNA library for pullulanase Hasnain, Hussain Zainab, Ngaini Q Science (General) QR Microbiology Pullulanases belong to a discrete class of debranching enzymes that are capable of hydrolysing the α-1,6-branches in starch. This work described the screening, identification and characterization of pullulanase gene from pea cDNA library. The screening process used degenerate primers based on the alignment of pullulanase genes from four plant species that has been previously identified. Initially, PCR using these primers for pullulanase on pea genomic DNA produced bands of about 600 bp, which was of approximately the expected size when compared to the sequence of pullulanase from spinach. Using this fragment as probe, screening of the pea cDNA library resulted in a full-length identification of a pullulanase gene with the size of 3.2 kb and encoded for 952 amino acids. Further analysis of the isolated clone, named PSPUL, from the screening process showed that it consists of 93 nucleotides in the 5’-untranslated region and 266 nucleotides in the 3’-untranslated region. The 3’-untranslated region contained three putative polyadenylation signals (AATAAT/A) located at 110 bp, 212 bp and 220 bp upstream of the last polyadenylation site. The predicted size for mature PSPUL peptides was 888 amino acids and calculation by PEPTIDESORT gave predicted molecular mass of 93 kDa. Three dimensional features of these enzymes showed that the pullulanase from pea contains eight regions of β-strand followed by eight regions of α-helix, which confirmed that it has the (β/α)8 characteristic of the α-amylase super family. 2011 Proceeding NonPeerReviewed text en http://ir.unimas.my/id/eprint/15580/1/Gene%20and%20structural.pdf Hasnain, Hussain and Zainab, Ngaini (2011) Gene and structural characterisation of pea cDNA library for pullulanase. In: 9th Malaysia Genetics Congress (MGC9). https://www.researchgate.net/publication/291341950_Gene_and_structural_characterisation_of_pea_cDNA_library_for_pullulanase
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)
QR Microbiology
spellingShingle Q Science (General)
QR Microbiology
Hasnain, Hussain
Zainab, Ngaini
Gene and structural characterisation of pea cDNA library for pullulanase
description Pullulanases belong to a discrete class of debranching enzymes that are capable of hydrolysing the α-1,6-branches in starch. This work described the screening, identification and characterization of pullulanase gene from pea cDNA library. The screening process used degenerate primers based on the alignment of pullulanase genes from four plant species that has been previously identified. Initially, PCR using these primers for pullulanase on pea genomic DNA produced bands of about 600 bp, which was of approximately the expected size when compared to the sequence of pullulanase from spinach. Using this fragment as probe, screening of the pea cDNA library resulted in a full-length identification of a pullulanase gene with the size of 3.2 kb and encoded for 952 amino acids. Further analysis of the isolated clone, named PSPUL, from the screening process showed that it consists of 93 nucleotides in the 5’-untranslated region and 266 nucleotides in the 3’-untranslated region. The 3’-untranslated region contained three putative polyadenylation signals (AATAAT/A) located at 110 bp, 212 bp and 220 bp upstream of the last polyadenylation site. The predicted size for mature PSPUL peptides was 888 amino acids and calculation by PEPTIDESORT gave predicted molecular mass of 93 kDa. Three dimensional features of these enzymes showed that the pullulanase from pea contains eight regions of β-strand followed by eight regions of α-helix, which confirmed that it has the (β/α)8 characteristic of the α-amylase super family.
format Proceeding
author Hasnain, Hussain
Zainab, Ngaini
author_facet Hasnain, Hussain
Zainab, Ngaini
author_sort Hasnain, Hussain
title Gene and structural characterisation of pea cDNA library for pullulanase
title_short Gene and structural characterisation of pea cDNA library for pullulanase
title_full Gene and structural characterisation of pea cDNA library for pullulanase
title_fullStr Gene and structural characterisation of pea cDNA library for pullulanase
title_full_unstemmed Gene and structural characterisation of pea cDNA library for pullulanase
title_sort gene and structural characterisation of pea cdna library for pullulanase
publishDate 2011
url http://ir.unimas.my/id/eprint/15580/1/Gene%20and%20structural.pdf
http://ir.unimas.my/id/eprint/15580/
https://www.researchgate.net/publication/291341950_Gene_and_structural_characterisation_of_pea_cDNA_library_for_pullulanase
_version_ 1724078487818469376
score 13.252575

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