Citation
Raja Noor Zaliha R. A. R., . and Lokman S., . and Mahiran B., . and Moohamad Ropaning S., . and Abu Bakar S., . (2010) Cloning and expression of industrial important thermostable amylase gene. [Proceedings Paper]
Abstract
Production of thermostable amylases was achieved using local strains. These strains were isolated from various hot springs with water temperatures ranging from 65C to 90C. However a quantitative test of the thermostable amylases using dinitrosalicylic acid DNSA method revealed inadequate level of the enzyme production 0.36 U/ml. Therefore molecular approaches such as cloning and expression were adopted to increase the amount of amylases. The thermostable amylase gene with approaximately 1.6 kb was successfully isolated. The gene was cloned into pET-32b vector and then transformed into E. coli BL21 expression host. The formation of clear zone on starch agar plate proved the presence of the recombinant amylases and thus indicated the success in cloning and expression of thermostable amylase gene. The level of the recombinant thermostable amylase after 24 h of induction time was higher as compared to the wild type 2.3 U/ml. Studies on the optimization of different concentrations of inducer IPTG as well as the post induction time are in progress.
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Abstract
Production of thermostable amylases was achieved using local strains. These strains were isolated from various hot springs with water temperatures ranging from 65C to 90C. However a quantitative test of the thermostable amylases using dinitrosalicylic acid DNSA method revealed inadequate level of the enzyme production 0.36 U/ml. Therefore molecular approaches such as cloning and expression were adopted to increase the amount of amylases. The thermostable amylase gene with approaximately 1.6 kb was successfully isolated. The gene was cloned into pET-32b vector and then transformed into E. coli BL21 expression host. The formation of clear zone on starch agar plate proved the presence of the recombinant amylases and thus indicated the success in cloning and expression of thermostable amylase gene. The level of the recombinant thermostable amylase after 24 h of induction time was higher as compared to the wild type 2.3 U/ml. Studies on the optimization of different concentrations of inducer IPTG as well as the post induction time are in progress.
Additional Metadata
| Item Type: | Proceedings Paper |
|---|---|
| Additional Information: | Available at Perpustakaan Sultan Abdul Samad Universiti Putra Malaysia 43400 UPM Serdang Selangor Malaysia. TP248.14 I61 2008 Call Number. |
| AGROVOC Term: | Amylases |
| AGROVOC Term: | Enzymes |
| AGROVOC Term: | Genes |
| AGROVOC Term: | Isolation of microorganisms |
| AGROVOC Term: | Gene cloning |
| AGROVOC Term: | Gene expression |
| AGROVOC Term: | Hot water treatment |
| AGROVOC Term: | genomics |
| AGROVOC Term: | DNA sequence |
| AGROVOC Term: | Enzyme activity |
| Geographical Term: | MALAYSIA |
| Depositing User: | Ms. Suzila Mohamad Kasim |
| Last Modified: | 24 Apr 2025 05:15 |
| URI: | http://webagris.upm.edu.my/id/eprint/13181 |
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