Cloning, extracellular expression and characterization of a predominant beta-CGTase from Bacillus sp G1 in E-coli
The cyclodextrin glucanotransferase (CGTase, EC 2.4.1.19) gene from Bacillus sp. G1 was successfully isolated and cloned into Escherichia coli. Analysis of the nucleotide sequence revealed the presence of an open reading frame of 2,109 bp and encoded a 674 amino acid protein. Purified CGTase exhibit...
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| Format: | Article |
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Springer Verlag
2008
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| author | Ong, Rui Min Goh, Kian Mau Mahadi, Nor Muhammad Hassan, Osman Raja Abdul Rahman, Raja Noor Zaliha Md. Illias, Rosli |
| author_facet | Ong, Rui Min Goh, Kian Mau Mahadi, Nor Muhammad Hassan, Osman Raja Abdul Rahman, Raja Noor Zaliha Md. Illias, Rosli |
| author_sort | Ong, Rui Min |
| collection | ePrints |
| description | The cyclodextrin glucanotransferase (CGTase, EC 2.4.1.19) gene from Bacillus sp. G1 was successfully isolated and cloned into Escherichia coli. Analysis of the nucleotide sequence revealed the presence of an open reading frame of 2,109 bp and encoded a 674 amino acid protein. Purified CGTase exhibited a molecular weight of 75 kDa and had optimum activity at pH 6 and 60C. Heterologous recombinant protein expression in E. coli is commonly problematic causing intracellular localization and formation of inactive inclusion bodies. This paper shows that the majority of CGTase was secreted into the medium due to the signal peptide of Bacillus sp. G1 that also works well in E. coli, leading to easier purification steps. When reacted with starch, CGTase G1 produced 90% beta-cyclodextrin (CD) and 10% gamma-CD. This enzyme also preferred the economical tapioca starch as a substrate, based on kinetics studies. Therefore, CGTase G1 could potentially serve as an industrial enzyme for the production of beta-CD. |
| first_indexed | 2024-03-05T18:23:32Z |
| format | Article |
| id | utm.eprints-12511 |
| institution | Universiti Teknologi Malaysia - ePrints |
| last_indexed | 2024-03-05T18:23:32Z |
| publishDate | 2008 |
| publisher | Springer Verlag |
| record_format | dspace |
| spelling | utm.eprints-125112018-03-15T01:42:59Z http://eprints.utm.my/12511/ Cloning, extracellular expression and characterization of a predominant beta-CGTase from Bacillus sp G1 in E-coli Ong, Rui Min Goh, Kian Mau Mahadi, Nor Muhammad Hassan, Osman Raja Abdul Rahman, Raja Noor Zaliha Md. Illias, Rosli Q Science (General) The cyclodextrin glucanotransferase (CGTase, EC 2.4.1.19) gene from Bacillus sp. G1 was successfully isolated and cloned into Escherichia coli. Analysis of the nucleotide sequence revealed the presence of an open reading frame of 2,109 bp and encoded a 674 amino acid protein. Purified CGTase exhibited a molecular weight of 75 kDa and had optimum activity at pH 6 and 60C. Heterologous recombinant protein expression in E. coli is commonly problematic causing intracellular localization and formation of inactive inclusion bodies. This paper shows that the majority of CGTase was secreted into the medium due to the signal peptide of Bacillus sp. G1 that also works well in E. coli, leading to easier purification steps. When reacted with starch, CGTase G1 produced 90% beta-cyclodextrin (CD) and 10% gamma-CD. This enzyme also preferred the economical tapioca starch as a substrate, based on kinetics studies. Therefore, CGTase G1 could potentially serve as an industrial enzyme for the production of beta-CD. Springer Verlag 2008-12 Article PeerReviewed Ong, Rui Min and Goh, Kian Mau and Mahadi, Nor Muhammad and Hassan, Osman and Raja Abdul Rahman, Raja Noor Zaliha and Md. Illias, Rosli (2008) Cloning, extracellular expression and characterization of a predominant beta-CGTase from Bacillus sp G1 in E-coli. Journal Of Industrial Microbiology & Biotechnology, 35 . pp. 1705-1714. ISSN 1367-5435 http://www.springerlink.com/content/u165261504231305/ DOI:10.1007/s10295-008-0462-2 |
| spellingShingle | Q Science (General) Ong, Rui Min Goh, Kian Mau Mahadi, Nor Muhammad Hassan, Osman Raja Abdul Rahman, Raja Noor Zaliha Md. Illias, Rosli Cloning, extracellular expression and characterization of a predominant beta-CGTase from Bacillus sp G1 in E-coli |
| title | Cloning, extracellular expression and characterization of a predominant beta-CGTase from Bacillus sp G1 in E-coli |
| title_full | Cloning, extracellular expression and characterization of a predominant beta-CGTase from Bacillus sp G1 in E-coli |
| title_fullStr | Cloning, extracellular expression and characterization of a predominant beta-CGTase from Bacillus sp G1 in E-coli |
| title_full_unstemmed | Cloning, extracellular expression and characterization of a predominant beta-CGTase from Bacillus sp G1 in E-coli |
| title_short | Cloning, extracellular expression and characterization of a predominant beta-CGTase from Bacillus sp G1 in E-coli |
| title_sort | cloning extracellular expression and characterization of a predominant beta cgtase from bacillus sp g1 in e coli |
| topic | Q Science (General) |
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