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


Citation

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

Abstract

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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Additional Metadata

Item Type: Article
Divisions: Faculty of Biotechnology and Biomolecular Sciences
Publisher: Academic Journals
Keywords: Bacillus sp. NR5 UPM; Cyclodextrin glycosyltransferase; Molecular cloning; Nucleotide sequence
Depositing User: Nur Farahin Ramli
Date Deposited: 24 Feb 2014 03:21
Last Modified: 17 Nov 2017 08:17
URI: http://psasir.upm.edu.my/id/eprint/24091
Statistic Details: View Download Statistic

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