Staff View: Protein engineering of selected residues from conserved sequence regions of a novel Anoxybacillus α-amylase

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...

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Main Authors:	Ranjani, Velayudhan, Janecek, Stefan, Chai, Kian Piaw, Shahir, Shafinaz, Raja Abdul Rahman, Raja Noor Zaliha, Chan, Kok Gan, Goh, Kian Mau
Format:	Article
Language:	English
Published:	Nature Publishing Group 2014
Online Access:	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 http://psasir.upm.edu.my/id/eprint/35093/
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spelling	my.upm.eprints.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
institution	Universiti Putra Malaysia
building	UPM Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	Universiti Putra Malaysia
content_source	UPM Institutional Repository
url_provider	http://psasir.upm.edu.my/
language	English
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.
format	Article
author	Ranjani, Velayudhan Janecek, Stefan Chai, Kian Piaw Shahir, Shafinaz Raja Abdul Rahman, Raja Noor Zaliha Chan, Kok Gan Goh, Kian Mau
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
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
title	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_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_sort	protein engineering of selected residues from conserved sequence regions of a novel anoxybacillus α-amylase
publisher	Nature Publishing Group
publishDate	2014
url	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 http://psasir.upm.edu.my/id/eprint/35093/
_version_	1643831350948528128
score	13.211869

Protein engineering of selected residues from conserved sequence regions of a novel Anoxybacillus α-amylase

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