Molecular cloning and extracellular expression of cyclodextrin glycosyltransferase gene from Bacillus sp. NR5 UPM

The cloning of a polymerase chain reaction (PCR) gene fragment from Bacillus sp. NR5 UPM isolated from the soil in Malaysia into an Escherichia coli expression vector was successfully carried out. Analysis of the nucleotide sequences revealed the presence of an open reading frame of 2112 bp which en...

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Main Authors: Ramli, Norhayati, Abd. Aziz, Suraini, Hassan, Mohd Ali, Mohammed Alitheen, Noorjahan Banu, Kamaruddin, Kamarulzaman, Ibrahim, Zoolhilmi
Format: Article
Language:English
Published: Academic Journals 2011
Online Access:http://psasir.upm.edu.my/id/eprint/24091/
http://psasir.upm.edu.my/id/eprint/24091/1/24091.pdf
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author Ramli, Norhayati
Abd. Aziz, Suraini
Hassan, Mohd Ali
Mohammed Alitheen, Noorjahan Banu
Kamaruddin, Kamarulzaman
Ibrahim, Zoolhilmi
author_facet Ramli, Norhayati
Abd. Aziz, Suraini
Hassan, Mohd Ali
Mohammed Alitheen, Noorjahan Banu
Kamaruddin, Kamarulzaman
Ibrahim, Zoolhilmi
author_sort Ramli, Norhayati
building UPM Institutional Repository
collection Online Access
description The cloning of a polymerase chain reaction (PCR) gene fragment from Bacillus sp. NR5 UPM isolated from the soil in Malaysia into an Escherichia coli expression vector was successfully carried out. Analysis of the nucleotide sequences revealed the presence of an open reading frame of 2112 bp which encoded a protein containing 704 amino acids with a putative molecular weight of 78.6 kDa. The deduced amino acids sequence showed about 98% homology with the CGTase from Bacillus sp. KC201. Compared to the wild type, the CGTase that was produced in E. coli cells only required one-fourth of culture time and neutral pH to produce CGTase. After 12 h of cultivation, the CGTase activity in the culture medium reached 29.6 U/ml, which was approximately 2.5-fold higher than the CGTase from the parental strain. The CGTase was produced extracellularly by E. coli (94%) indicating the signal peptide was functional in E. coli.
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spelling upm-240912017-11-17T08:17:10Z http://psasir.upm.edu.my/id/eprint/24091/ Molecular cloning and extracellular expression of cyclodextrin glycosyltransferase gene from Bacillus sp. NR5 UPM Ramli, Norhayati Abd. Aziz, Suraini Hassan, Mohd Ali Mohammed Alitheen, Noorjahan Banu Kamaruddin, Kamarulzaman Ibrahim, Zoolhilmi The cloning of a polymerase chain reaction (PCR) gene fragment from Bacillus sp. NR5 UPM isolated from the soil in Malaysia into an Escherichia coli expression vector was successfully carried out. Analysis of the nucleotide sequences revealed the presence of an open reading frame of 2112 bp which encoded a protein containing 704 amino acids with a putative molecular weight of 78.6 kDa. The deduced amino acids sequence showed about 98% homology with the CGTase from Bacillus sp. KC201. Compared to the wild type, the CGTase that was produced in E. coli cells only required one-fourth of culture time and neutral pH to produce CGTase. After 12 h of cultivation, the CGTase activity in the culture medium reached 29.6 U/ml, which was approximately 2.5-fold higher than the CGTase from the parental strain. The CGTase was produced extracellularly by E. coli (94%) indicating the signal peptide was functional in E. coli. Academic Journals 2011 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/24091/1/24091.pdf Ramli, Norhayati and Abd. Aziz, Suraini and Hassan, Mohd Ali and Mohammed Alitheen, Noorjahan Banu and Kamaruddin, Kamarulzaman and Ibrahim, Zoolhilmi (2011) Molecular cloning and extracellular expression of cyclodextrin glycosyltransferase gene from Bacillus sp. NR5 UPM. African Journal of Microbiology Research, 5 (21). art. no. 3BF03B713165. pp. 3475-3482. ISSN 1996-0808 http://www.academicjournals.org/journal/AJMR/article-abstract/3BF03B713165
spellingShingle Ramli, Norhayati
Abd. Aziz, Suraini
Hassan, Mohd Ali
Mohammed Alitheen, Noorjahan Banu
Kamaruddin, Kamarulzaman
Ibrahim, Zoolhilmi
Molecular cloning and extracellular expression of cyclodextrin glycosyltransferase gene from Bacillus sp. NR5 UPM
title Molecular cloning and extracellular expression of cyclodextrin glycosyltransferase gene from Bacillus sp. NR5 UPM
title_full Molecular cloning and extracellular expression of cyclodextrin glycosyltransferase gene from Bacillus sp. NR5 UPM
title_fullStr Molecular cloning and extracellular expression of cyclodextrin glycosyltransferase gene from Bacillus sp. NR5 UPM
title_full_unstemmed Molecular cloning and extracellular expression of cyclodextrin glycosyltransferase gene from Bacillus sp. NR5 UPM
title_short Molecular cloning and extracellular expression of cyclodextrin glycosyltransferase gene from Bacillus sp. NR5 UPM
title_sort molecular cloning and extracellular expression of cyclodextrin glycosyltransferase gene from bacillus sp. nr5 upm
url http://psasir.upm.edu.my/id/eprint/24091/
http://psasir.upm.edu.my/id/eprint/24091/
http://psasir.upm.edu.my/id/eprint/24091/1/24091.pdf