Protein engineering of selected residues from conserved sequence regions of a novel Anoxybacillus α-amylase
The α-amylases from Anoxybacillus species (ASKA and ADTA), Bacillus aquimaris (BaqA) and Geobacillus thermoleovorans (GTA, Pizzo and GtamyII) were proposed as a novel group of the α-amylase family GH13. An ASKA yielding a high percentage of maltose upon its reaction on starch was chosen as a model t...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Publishing Group
2014
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| Online Access: | http://psasir.upm.edu.my/id/eprint/35093/ http://psasir.upm.edu.my/id/eprint/35093/1/Protein%20engineering%20of%20selected%20residues%20from%20conserved%20sequence%20regions%20of%20a%20novel%20Anoxybacillus%20%CE%B1-amylase.pdf |
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| author | Ranjani, Velayudhan Janecek, Stefan Chai, Kian Piaw Shahir, Shafinaz Raja Abdul Rahman, Raja Noor Zaliha Chan, Kok Gan Goh, Kian Mau |
| author_facet | Ranjani, Velayudhan Janecek, Stefan Chai, Kian Piaw Shahir, Shafinaz Raja Abdul Rahman, Raja Noor Zaliha Chan, Kok Gan Goh, Kian Mau |
| author_sort | Ranjani, Velayudhan |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | The α-amylases from Anoxybacillus species (ASKA and ADTA), Bacillus aquimaris (BaqA) and Geobacillus thermoleovorans (GTA, Pizzo and GtamyII) were proposed as a novel group of the α-amylase family GH13. An ASKA yielding a high percentage of maltose upon its reaction on starch was chosen as a model to study the residues responsible for the biochemical properties. Four residues from conserved sequence regions (CSRs) were thus selected, and the mutants F113V (CSR-I), Y187F and L189I (CSR-II) and A161D (CSR-V) were characterised. Few changes in the optimum reaction temperature and pH were observed for all mutants. Whereas the Y187F (t1/2 43 h) and L189I (t1/2 36 h) mutants had a lower thermostability at 65°C than the native ASKA (t1/2 48 h), the mutants F113V and A161D exhibited an improved t1/2 of 51 h and 53 h, respectively. Among the mutants, only the A161D had a specific activity, kcat and kcat/Km higher (1.23-, 1.17- and 2.88-times, respectively) than the values determined for the ASKA. The replacement of the Ala-161 in the CSR-V with an aspartic acid also caused a significant reduction in the ratio of maltose formed. This finding suggests the Ala-161 may contribute to the high maltose production of the ASKA. |
| first_indexed | 2025-11-15T09:26:52Z |
| format | Article |
| id | upm-35093 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T09:26:52Z |
| publishDate | 2014 |
| publisher | Nature Publishing Group |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-350932016-09-28T07:09:23Z http://psasir.upm.edu.my/id/eprint/35093/ Protein engineering of selected residues from conserved sequence regions of a novel Anoxybacillus α-amylase Ranjani, Velayudhan Janecek, Stefan Chai, Kian Piaw Shahir, Shafinaz Raja Abdul Rahman, Raja Noor Zaliha Chan, Kok Gan Goh, Kian Mau The α-amylases from Anoxybacillus species (ASKA and ADTA), Bacillus aquimaris (BaqA) and Geobacillus thermoleovorans (GTA, Pizzo and GtamyII) were proposed as a novel group of the α-amylase family GH13. An ASKA yielding a high percentage of maltose upon its reaction on starch was chosen as a model to study the residues responsible for the biochemical properties. Four residues from conserved sequence regions (CSRs) were thus selected, and the mutants F113V (CSR-I), Y187F and L189I (CSR-II) and A161D (CSR-V) were characterised. Few changes in the optimum reaction temperature and pH were observed for all mutants. Whereas the Y187F (t1/2 43 h) and L189I (t1/2 36 h) mutants had a lower thermostability at 65°C than the native ASKA (t1/2 48 h), the mutants F113V and A161D exhibited an improved t1/2 of 51 h and 53 h, respectively. Among the mutants, only the A161D had a specific activity, kcat and kcat/Km higher (1.23-, 1.17- and 2.88-times, respectively) than the values determined for the ASKA. The replacement of the Ala-161 in the CSR-V with an aspartic acid also caused a significant reduction in the ratio of maltose formed. This finding suggests the Ala-161 may contribute to the high maltose production of the ASKA. Nature Publishing Group 2014-07-28 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/35093/1/Protein%20engineering%20of%20selected%20residues%20from%20conserved%20sequence%20regions%20of%20a%20novel%20Anoxybacillus%20%CE%B1-amylase.pdf Ranjani, Velayudhan and Janecek, Stefan and Chai, Kian Piaw and Shahir, Shafinaz and Raja Abdul Rahman, Raja Noor Zaliha and Chan, Kok Gan and Goh, Kian Mau (2014) Protein engineering of selected residues from conserved sequence regions of a novel Anoxybacillus α-amylase. Scientific Reports, 4. art. no. 5850. pp. 1-8. ISSN 2045-2322 10.1038/srep05850 |
| spellingShingle | Ranjani, Velayudhan Janecek, Stefan Chai, Kian Piaw Shahir, Shafinaz Raja Abdul Rahman, Raja Noor Zaliha Chan, Kok Gan Goh, Kian Mau Protein engineering of selected residues from conserved sequence regions of a novel Anoxybacillus α-amylase |
| title | Protein engineering of selected residues from conserved sequence regions of a novel Anoxybacillus α-amylase |
| title_full | Protein engineering of selected residues from conserved sequence regions of a novel Anoxybacillus α-amylase |
| title_fullStr | Protein engineering of selected residues from conserved sequence regions of a novel Anoxybacillus α-amylase |
| title_full_unstemmed | Protein engineering of selected residues from conserved sequence regions of a novel Anoxybacillus α-amylase |
| title_short | Protein engineering of selected residues from conserved sequence regions of a novel Anoxybacillus α-amylase |
| title_sort | protein engineering of selected residues from conserved sequence regions of a novel anoxybacillus α-amylase |
| url | http://psasir.upm.edu.my/id/eprint/35093/ http://psasir.upm.edu.my/id/eprint/35093/ http://psasir.upm.edu.my/id/eprint/35093/1/Protein%20engineering%20of%20selected%20residues%20from%20conserved%20sequence%20regions%20of%20a%20novel%20Anoxybacillus%20%CE%B1-amylase.pdf |