Cloning and expression of industrial important thermostable amylase gene
Production of thermostable amylases was achieved using local strains. These strains were isolated from various hot springs with water temperatures ranging from 65°C to 90°C. However, a quantitative test of the thermostable amylases using dinitrosalicylic acid (DNSA) method revealed inadequate level...
| Main Authors: | , , , , |
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| Format: | Conference or Workshop Item |
| Language: | English |
| Published: |
Sarawak Biodiversity Centre
2008
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| Online Access: | http://psasir.upm.edu.my/id/eprint/64276/1/PROCEEDING_100719-13.pdf |
| _version_ | 1796977866032807936 |
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| author | Sulong, Moohamad Ropaning Shamsudin, Lokman Salleh, Abu Bakar Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran |
| author_facet | Sulong, Moohamad Ropaning Shamsudin, Lokman Salleh, Abu Bakar Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran |
| author_sort | Sulong, Moohamad Ropaning |
| collection | UPM |
| description | Production of thermostable amylases was achieved using local strains. These strains were isolated from various hot springs with water temperatures ranging from 65°C to 90°C. 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. |
| first_indexed | 2024-03-06T09:46:19Z |
| format | Conference or Workshop Item |
| id | upm.eprints-64276 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T09:46:19Z |
| publishDate | 2008 |
| publisher | Sarawak Biodiversity Centre |
| record_format | dspace |
| spelling | upm.eprints-642762018-07-04T02:31:39Z http://psasir.upm.edu.my/id/eprint/64276/ Cloning and expression of industrial important thermostable amylase gene Sulong, Moohamad Ropaning Shamsudin, Lokman Salleh, Abu Bakar Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Production of thermostable amylases was achieved using local strains. These strains were isolated from various hot springs with water temperatures ranging from 65°C to 90°C. 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. Sarawak Biodiversity Centre 2008 Conference or Workshop Item PeerReviewed text en http://psasir.upm.edu.my/id/eprint/64276/1/PROCEEDING_100719-13.pdf Sulong, Moohamad Ropaning and Shamsudin, Lokman and Salleh, Abu Bakar and Raja Abdul Rahman, Raja Noor Zaliha and Basri, Mahiran (2008) Cloning and expression of industrial important thermostable amylase gene. In: Biodiversity and Biotechnology Symposium 2008, 19-21 Nov. 2008, Kuching, Sarawak, Malaysia. (pp. 399-402). |
| spellingShingle | Sulong, Moohamad Ropaning Shamsudin, Lokman Salleh, Abu Bakar Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Cloning and expression of industrial important thermostable amylase gene |
| title | Cloning and expression of industrial important thermostable amylase gene |
| title_full | Cloning and expression of industrial important thermostable amylase gene |
| title_fullStr | Cloning and expression of industrial important thermostable amylase gene |
| title_full_unstemmed | Cloning and expression of industrial important thermostable amylase gene |
| title_short | Cloning and expression of industrial important thermostable amylase gene |
| title_sort | cloning and expression of industrial important thermostable amylase gene |
| url | http://psasir.upm.edu.my/id/eprint/64276/1/PROCEEDING_100719-13.pdf |
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