Heterologous, Expression, and Characterization of Thermostable Glucoamylase Derived from Aspergillus flavus NSH9 in Pichia pastoris
A novel thermostable glucoamylase cDNA without starch binding domain (SBD) of Aspergillus flavus NSH9 was successfully identified, isolated, and overexpressed in Pichia pastoris GS115.The complete open reading frame of glucoamylase from Aspergillus flavus NSH9 was identified by employing PCR that...
Saved in:
Main Authors: | , , , , , , |
---|---|
Format: | E-Article |
Language: | English |
Published: |
Hindawi Publishing Corporation
2016
|
Subjects: | |
Online Access: | http://ir.unimas.my/id/eprint/15732/1/Heterologous%2C%20Expression%2C%20and%20Characterization%20of%20%28abstract%29.pdf http://ir.unimas.my/id/eprint/15732/ https://www.hindawi.com/journals/bmri/2016/5962028/ http://dx.doi.org/10.1155/2016/5962028 |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
id |
my.unimas.ir.15732 |
---|---|
record_format |
eprints |
spelling |
my.unimas.ir.157322017-03-30T06:41:12Z http://ir.unimas.my/id/eprint/15732/ Heterologous, Expression, and Characterization of Thermostable Glucoamylase Derived from Aspergillus flavus NSH9 in Pichia pastoris Kazi Muhammad, Rezaul Karim Ahmad, Husaini Md. Anowar, Hossain Ngieng, Ngui Sing Fazia, Mohd Sinang Mohd Hasnain, Md Hussain Hairul Azman, Roslan Q Science (General) A novel thermostable glucoamylase cDNA without starch binding domain (SBD) of Aspergillus flavus NSH9 was successfully identified, isolated, and overexpressed in Pichia pastoris GS115.The complete open reading frame of glucoamylase from Aspergillus flavus NSH9 was identified by employing PCR that encodes 493 amino acids lacking in the SBD. The first 17 amino acids were presumed to be a signal peptide.The cDNA was cloned into Pichia pastoris and the highest expression of recombinant glucoamylase (rGA) was observed after 8 days of incubation period with 1% methanol. The molecular weight of the purified rGA was about 78 kDa and exhibited optimum catalytic activity at pH 5.0 and temperature of 70∘C. The enzyme was stable at higher temperature with 50% of residual activity observed after 20 min at 90∘C and 100∘C. Low concentration of metal (Mg++, Fe++, Zn++, Cu++, and Pb++) had positive effect on rGA activity.This rGA has the potential for use and application in the saccharification steps, due to its thermostability, in the starch processing industries. Hindawi Publishing Corporation 2016 E-Article PeerReviewed text en http://ir.unimas.my/id/eprint/15732/1/Heterologous%2C%20Expression%2C%20and%20Characterization%20of%20%28abstract%29.pdf Kazi Muhammad, Rezaul Karim and Ahmad, Husaini and Md. Anowar, Hossain and Ngieng, Ngui Sing and Fazia, Mohd Sinang and Mohd Hasnain, Md Hussain and Hairul Azman, Roslan (2016) Heterologous, Expression, and Characterization of Thermostable Glucoamylase Derived from Aspergillus flavus NSH9 in Pichia pastoris. BioMed Research International, 2016. ISSN 23146133 https://www.hindawi.com/journals/bmri/2016/5962028/ http://dx.doi.org/10.1155/2016/5962028 |
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) Kazi Muhammad, Rezaul Karim Ahmad, Husaini Md. Anowar, Hossain Ngieng, Ngui Sing Fazia, Mohd Sinang Mohd Hasnain, Md Hussain Hairul Azman, Roslan Heterologous, Expression, and Characterization of Thermostable Glucoamylase Derived from Aspergillus flavus NSH9 in Pichia pastoris |
description |
A novel thermostable glucoamylase cDNA without starch binding domain (SBD) of Aspergillus flavus NSH9 was successfully
identified, isolated, and overexpressed in Pichia pastoris GS115.The complete open reading frame of glucoamylase from Aspergillus
flavus NSH9 was identified by employing PCR that encodes 493 amino acids lacking in the SBD. The first 17 amino acids were
presumed to be a signal peptide.The cDNA was cloned into Pichia pastoris and the highest expression of recombinant glucoamylase
(rGA) was observed after 8 days of incubation period with 1% methanol. The molecular weight of the purified rGA was about
78 kDa and exhibited optimum catalytic activity at pH 5.0 and temperature of 70∘C. The enzyme was stable at higher temperature
with 50% of residual activity observed after 20 min at 90∘C and 100∘C. Low concentration of metal (Mg++, Fe++, Zn++, Cu++, and
Pb++) had positive effect on rGA activity.This rGA has the potential for use and application in the saccharification steps, due to its
thermostability, in the starch processing industries. |
format |
E-Article |
author |
Kazi Muhammad, Rezaul Karim Ahmad, Husaini Md. Anowar, Hossain Ngieng, Ngui Sing Fazia, Mohd Sinang Mohd Hasnain, Md Hussain Hairul Azman, Roslan |
author_facet |
Kazi Muhammad, Rezaul Karim Ahmad, Husaini Md. Anowar, Hossain Ngieng, Ngui Sing Fazia, Mohd Sinang Mohd Hasnain, Md Hussain Hairul Azman, Roslan |
author_sort |
Kazi Muhammad, Rezaul Karim |
title |
Heterologous, Expression, and Characterization of Thermostable Glucoamylase Derived from Aspergillus flavus NSH9 in Pichia pastoris |
title_short |
Heterologous, Expression, and Characterization of Thermostable Glucoamylase Derived from Aspergillus flavus NSH9 in Pichia pastoris |
title_full |
Heterologous, Expression, and Characterization of Thermostable Glucoamylase Derived from Aspergillus flavus NSH9 in Pichia pastoris |
title_fullStr |
Heterologous, Expression, and Characterization of Thermostable Glucoamylase Derived from Aspergillus flavus NSH9 in Pichia pastoris |
title_full_unstemmed |
Heterologous, Expression, and Characterization of Thermostable Glucoamylase Derived from Aspergillus flavus NSH9 in Pichia pastoris |
title_sort |
heterologous, expression, and characterization of thermostable glucoamylase derived from aspergillus flavus nsh9 in pichia pastoris |
publisher |
Hindawi Publishing Corporation |
publishDate |
2016 |
url |
http://ir.unimas.my/id/eprint/15732/1/Heterologous%2C%20Expression%2C%20and%20Characterization%20of%20%28abstract%29.pdf http://ir.unimas.my/id/eprint/15732/ https://www.hindawi.com/journals/bmri/2016/5962028/ http://dx.doi.org/10.1155/2016/5962028 |
_version_ |
1644512218341965824 |
score |
13.160551 |