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 en...
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| Format: | Article |
| Language: | English |
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Hidawi Publishing Coperation
2016
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| Online Access: | http://ir.unimas.my/id/eprint/12764/ http://ir.unimas.my/id/eprint/12764/1/Kazi%20Muhammad.pdf |
| _version_ | 1848837270556114944 |
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| 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 |
| building | UNIMAS Institutional Repository |
| collection | Online Access |
| 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. |
| first_indexed | 2025-11-15T06:37:00Z |
| format | Article |
| id | unimas-12764 |
| institution | Universiti Malaysia Sarawak |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T06:37:00Z |
| publishDate | 2016 |
| publisher | Hidawi Publishing Coperation |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | unimas-127642022-02-07T06:14:31Z http://ir.unimas.my/id/eprint/12764/ 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) QD Chemistry 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. Hidawi Publishing Coperation 2016 Article PeerReviewed text en http://ir.unimas.my/id/eprint/12764/1/Kazi%20Muhammad.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 2314-6141 https://www.scopus.com/inward/record.uri?eid=2-s2.0-84982796344&partnerID=40&md5=9a99c00cfdab1191cf4cd5bb9858b1e3 http://dx.doi.org/10.1155/2016/5962028 |
| spellingShingle | Q Science (General) QD Chemistry 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 |
| title | 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_short | 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 |
| topic | Q Science (General) QD Chemistry |
| url | http://ir.unimas.my/id/eprint/12764/ http://ir.unimas.my/id/eprint/12764/ http://ir.unimas.my/id/eprint/12764/ http://ir.unimas.my/id/eprint/12764/1/Kazi%20Muhammad.pdf |