In silico analysis and truncation studies of the c terminal alpha amylase from anoxybacillus species

Thermostable and alkalitolerant a-amylase from Anoxybacillus sp. SK3-4 (ASKA) yields high level of maltose (69%) from starch hydrolysis. A putative raw starch binding domain was found in the C-terminal domain of ASKA. Starch binding domain (SBD) plays a role in hydrolyzing insoluble starch which enh...

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Main Author:	Sea, Lillian Shun Yi
Format:	Thesis
Language:	English
Published:	2015
Subjects:	R Medicine (General)
Online Access:	http://eprints.utm.my/id/eprint/54653/25/LillianSeaShunYiMFBME2015.pdf
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spelling	my-utm-ep.546532020-11-03T07:21:56Z In silico analysis and truncation studies of the c terminal alpha amylase from anoxybacillus species 2015-02 Sea, Lillian Shun Yi R Medicine (General) Thermostable and alkalitolerant a-amylase from Anoxybacillus sp. SK3-4 (ASKA) yields high level of maltose (69%) from starch hydrolysis. A putative raw starch binding domain was found in the C-terminal domain of ASKA. Starch binding domain (SBD) plays a role in hydrolyzing insoluble starch which enhances the amylolytic rate. The gene that encodes the ASKA is 1,518 bp. The present study involved in silico analyses and protein mutagenesis of two C-terminal truncated variants, namely mutant ?50 and ?100. The respective mutants had an elimination of 81 and 112 amino acids from C-terminal end of ASKA. Firstly mutagenesis was made in in silico structure. Bioinformatics analysis revealed that the eight ß-strand forms as an open-sided distorted ß-sandwich fold which of putative domain C. Using Neighbor-Joining (NJ) method, the phylogenetic analysis of the domain C had confirmed that starch binding region of ASKA is the non-catalytic domain C and not the typical SBD of CBM families. Actual gene truncation was constructed via protein engineering approach and the truncated genes were expressed in Escherichia coli, under the promoter of T7. However, both recombinant proteins exhibited no activity with soluble starch as a result of inclusion bodies aggregation; despite several optimization on expression conditions were made. 2015-02 Thesis http://eprints.utm.my/id/eprint/54653/ http://eprints.utm.my/id/eprint/54653/25/LillianSeaShunYiMFBME2015.pdf application/pdf en public http://dms.library.utm.my:8080/vital/access/manager/Repository/vital:86580 masters Universiti Teknologi Malaysia, Faculty of Biosciences and Medical Engineering Faculty of Biosciences and Medical Engineering
institution	Universiti Teknologi Malaysia
collection	UTM Institutional Repository
language	English
topic	R Medicine (General)
spellingShingle	R Medicine (General) Sea, Lillian Shun Yi In silico analysis and truncation studies of the c terminal alpha amylase from anoxybacillus species
description	Thermostable and alkalitolerant a-amylase from Anoxybacillus sp. SK3-4 (ASKA) yields high level of maltose (69%) from starch hydrolysis. A putative raw starch binding domain was found in the C-terminal domain of ASKA. Starch binding domain (SBD) plays a role in hydrolyzing insoluble starch which enhances the amylolytic rate. The gene that encodes the ASKA is 1,518 bp. The present study involved in silico analyses and protein mutagenesis of two C-terminal truncated variants, namely mutant ?50 and ?100. The respective mutants had an elimination of 81 and 112 amino acids from C-terminal end of ASKA. Firstly mutagenesis was made in in silico structure. Bioinformatics analysis revealed that the eight ß-strand forms as an open-sided distorted ß-sandwich fold which of putative domain C. Using Neighbor-Joining (NJ) method, the phylogenetic analysis of the domain C had confirmed that starch binding region of ASKA is the non-catalytic domain C and not the typical SBD of CBM families. Actual gene truncation was constructed via protein engineering approach and the truncated genes were expressed in Escherichia coli, under the promoter of T7. However, both recombinant proteins exhibited no activity with soluble starch as a result of inclusion bodies aggregation; despite several optimization on expression conditions were made.
format	Thesis
qualification_level	Master's degree
author	Sea, Lillian Shun Yi
author_facet	Sea, Lillian Shun Yi
author_sort	Sea, Lillian Shun Yi
title	In silico analysis and truncation studies of the c terminal alpha amylase from anoxybacillus species
title_short	In silico analysis and truncation studies of the c terminal alpha amylase from anoxybacillus species
title_full	In silico analysis and truncation studies of the c terminal alpha amylase from anoxybacillus species
title_fullStr	In silico analysis and truncation studies of the c terminal alpha amylase from anoxybacillus species
title_full_unstemmed	In silico analysis and truncation studies of the c terminal alpha amylase from anoxybacillus species
title_sort	in silico analysis and truncation studies of the c terminal alpha amylase from anoxybacillus species
granting_institution	Universiti Teknologi Malaysia, Faculty of Biosciences and Medical Engineering
granting_department	Faculty of Biosciences and Medical Engineering
publishDate	2015
url	http://eprints.utm.my/id/eprint/54653/25/LillianSeaShunYiMFBME2015.pdf
_version_	1747817697281835008

In silico analysis and truncation studies of the c terminal alpha amylase from anoxybacillus species

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