Expression and characterization of thermostable glycogen branching enzyme from Geobacillus mahadia Geo-05
The glycogen branching enzyme (EC 2.4.1.18), which catalyses the formation of α-1,6-glycosidic branch points in glycogen structure, is often used to enhance the nutritional value and quality of food and beverages. In order to be applicable in industries, enzymes that are stable and active at high te...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
PeerJ
2016
|
| Subjects: | |
| Online Access: | http://psasir.upm.edu.my/id/eprint/54244/1/Expression%20and%20characterization%20of%20thermostable%20glycogen%20branching%20enzyme.pdf |
| _version_ | 1825931000423645184 |
|---|---|
| author | Mohtar, Nur Syazwani Abdul Rahman, Mohd Basyaruddin Raja Abd Rahman, Raja Noor Zaliha Leow, Thean Chor Salleh, Abu Bakar Mat Isa, Mohd Noor |
| author_facet | Mohtar, Nur Syazwani Abdul Rahman, Mohd Basyaruddin Raja Abd Rahman, Raja Noor Zaliha Leow, Thean Chor Salleh, Abu Bakar Mat Isa, Mohd Noor |
| author_sort | Mohtar, Nur Syazwani |
| collection | UPM |
| description | The glycogen branching enzyme (EC 2.4.1.18), which catalyses the formation of α-1,6-glycosidic branch points in glycogen structure, is often used to enhance the nutritional value and quality of food and beverages. In order to be applicable in industries, enzymes that are stable and active at high temperature are much desired. Using genome mining, the nucleotide sequence of the branching enzyme gene (glgB) was extracted from the Geobacillus mahadia Geo-05 genome sequence provided by the Malaysia Genome Institute. The size of the gene is 2013 bp, and the theoretical molecular weight of the protein is 78.43 kDa. The gene sequence was then used to predict the thermostability, function and the three dimensional structure of the enzyme. The gene was cloned and overexpressed in E. coli to verify the predicted result experimentally. The purified enzyme was used to study the effect of temperature and pH on enzyme activity and stability, and the inhibitory effect by metal ion on enzyme activity. This thermostable glycogen branching enzyme was found to be most active at 55 °C, and the half-life at 60 °C and 70 °C was 24 h and 5 h, respectively. From this research, a thermostable glycogen branching enzyme was successfully isolated from Geobacillus mahadia Geo-05 by genome mining together with molecular biology technique. |
| first_indexed | 2024-03-06T09:20:06Z |
| format | Article |
| id | upm.eprints-54244 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T09:20:06Z |
| publishDate | 2016 |
| publisher | PeerJ |
| record_format | dspace |
| spelling | upm.eprints-542442018-03-08T04:32:20Z http://psasir.upm.edu.my/id/eprint/54244/ Expression and characterization of thermostable glycogen branching enzyme from Geobacillus mahadia Geo-05 Mohtar, Nur Syazwani Abdul Rahman, Mohd Basyaruddin Raja Abd Rahman, Raja Noor Zaliha Leow, Thean Chor Salleh, Abu Bakar Mat Isa, Mohd Noor The glycogen branching enzyme (EC 2.4.1.18), which catalyses the formation of α-1,6-glycosidic branch points in glycogen structure, is often used to enhance the nutritional value and quality of food and beverages. In order to be applicable in industries, enzymes that are stable and active at high temperature are much desired. Using genome mining, the nucleotide sequence of the branching enzyme gene (glgB) was extracted from the Geobacillus mahadia Geo-05 genome sequence provided by the Malaysia Genome Institute. The size of the gene is 2013 bp, and the theoretical molecular weight of the protein is 78.43 kDa. The gene sequence was then used to predict the thermostability, function and the three dimensional structure of the enzyme. The gene was cloned and overexpressed in E. coli to verify the predicted result experimentally. The purified enzyme was used to study the effect of temperature and pH on enzyme activity and stability, and the inhibitory effect by metal ion on enzyme activity. This thermostable glycogen branching enzyme was found to be most active at 55 °C, and the half-life at 60 °C and 70 °C was 24 h and 5 h, respectively. From this research, a thermostable glycogen branching enzyme was successfully isolated from Geobacillus mahadia Geo-05 by genome mining together with molecular biology technique. PeerJ 2016 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/54244/1/Expression%20and%20characterization%20of%20thermostable%20glycogen%20branching%20enzyme.pdf Mohtar, Nur Syazwani and Abdul Rahman, Mohd Basyaruddin and Raja Abd Rahman, Raja Noor Zaliha and Leow, Thean Chor and Salleh, Abu Bakar and Mat Isa, Mohd Noor (2016) Expression and characterization of thermostable glycogen branching enzyme from Geobacillus mahadia Geo-05. PeerJ, 4. pp. 1-12. ISSN 2167-8359 https://peerj.com/articles/2714/ 1-4-alpha-glucan branching enzyme; His-patch thioredoxin; Geobacillus sp; Glycogen branching enzyme; Genome mining 10.7717/peerj.2714 |
| spellingShingle | 1-4-alpha-glucan branching enzyme; His-patch thioredoxin; Geobacillus sp; Glycogen branching enzyme; Genome mining Mohtar, Nur Syazwani Abdul Rahman, Mohd Basyaruddin Raja Abd Rahman, Raja Noor Zaliha Leow, Thean Chor Salleh, Abu Bakar Mat Isa, Mohd Noor Expression and characterization of thermostable glycogen branching enzyme from Geobacillus mahadia Geo-05 |
| title | Expression and characterization of thermostable glycogen branching enzyme from Geobacillus mahadia Geo-05 |
| title_full | Expression and characterization of thermostable glycogen branching enzyme from Geobacillus mahadia Geo-05 |
| title_fullStr | Expression and characterization of thermostable glycogen branching enzyme from Geobacillus mahadia Geo-05 |
| title_full_unstemmed | Expression and characterization of thermostable glycogen branching enzyme from Geobacillus mahadia Geo-05 |
| title_short | Expression and characterization of thermostable glycogen branching enzyme from Geobacillus mahadia Geo-05 |
| title_sort | expression and characterization of thermostable glycogen branching enzyme from geobacillus mahadia geo 05 |
| topic | 1-4-alpha-glucan branching enzyme; His-patch thioredoxin; Geobacillus sp; Glycogen branching enzyme; Genome mining |
| url | http://psasir.upm.edu.my/id/eprint/54244/1/Expression%20and%20characterization%20of%20thermostable%20glycogen%20branching%20enzyme.pdf |
| work_keys_str_mv | AT mohtarnursyazwani expressionandcharacterizationofthermostableglycogenbranchingenzymefromgeobacillusmahadiageo05 AT abdulrahmanmohdbasyaruddin expressionandcharacterizationofthermostableglycogenbranchingenzymefromgeobacillusmahadiageo05 AT rajaabdrahmanrajanoorzaliha expressionandcharacterizationofthermostableglycogenbranchingenzymefromgeobacillusmahadiageo05 AT leowtheanchor expressionandcharacterizationofthermostableglycogenbranchingenzymefromgeobacillusmahadiageo05 AT sallehabubakar expressionandcharacterizationofthermostableglycogenbranchingenzymefromgeobacillusmahadiageo05 AT matisamohdnoor expressionandcharacterizationofthermostableglycogenbranchingenzymefromgeobacillusmahadiageo05 |