Molecular investigation of a gene encoding organic solvent-tolerant alkaline protease from Pseudomonas aeruginosa strain K
A gene encoding an organic solvent-stable protease was amplified from Pseudomonas aeruginosa strain K by polymerase chain reaction using consensus primers based on multiple sequence alignment of alkaline and metalloprotease genes from Pseudomonas species. The gene, which consisted of 1440 bp nucleot...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wiley-VCH Verlag GmbH & Co. KGaA
2010
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| Online Access: | http://psasir.upm.edu.my/id/eprint/15709/1/15709.pdf |
| _version_ | 1796968903475200000 |
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| author | Raja Abdul Rahman, Raja Noor Zaliha Lee, Poh Geok Wong, Chee Fah Basri, Mahiran Salleh, Abu Bakar |
| author_facet | Raja Abdul Rahman, Raja Noor Zaliha Lee, Poh Geok Wong, Chee Fah Basri, Mahiran Salleh, Abu Bakar |
| author_sort | Raja Abdul Rahman, Raja Noor Zaliha |
| collection | UPM |
| description | A gene encoding an organic solvent-stable protease was amplified from Pseudomonas aeruginosa strain K by polymerase chain reaction using consensus primers based on multiple sequence alignment of alkaline and metalloprotease genes from Pseudomonas species. The gene, which consisted of 1440 bp nucleotides and deduced 479 amino acid residues, was successfully expressed in pGEX-4T-1 expression system in the presence of 1.0 mM IPTG, after an incubation of 6 h at 37 degrees C. Under these conditions, the recombinant strain K protease was, subsequently, released into the periplasm of E. coli BL21 (DE3) with an optimum proteolytic activity detected at 1.0112 U/ml. To date, this is the first reported expression of alkaline protease (aprA) with such remarkable property in Escherichia coli. |
| first_indexed | 2024-03-06T07:35:09Z |
| format | Article |
| id | upm.eprints-15709 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T07:35:09Z |
| publishDate | 2010 |
| publisher | Wiley-VCH Verlag GmbH & Co. KGaA |
| record_format | dspace |
| spelling | upm.eprints-157092016-09-26T04:57:36Z http://psasir.upm.edu.my/id/eprint/15709/ Molecular investigation of a gene encoding organic solvent-tolerant alkaline protease from Pseudomonas aeruginosa strain K Raja Abdul Rahman, Raja Noor Zaliha Lee, Poh Geok Wong, Chee Fah Basri, Mahiran Salleh, Abu Bakar A gene encoding an organic solvent-stable protease was amplified from Pseudomonas aeruginosa strain K by polymerase chain reaction using consensus primers based on multiple sequence alignment of alkaline and metalloprotease genes from Pseudomonas species. The gene, which consisted of 1440 bp nucleotides and deduced 479 amino acid residues, was successfully expressed in pGEX-4T-1 expression system in the presence of 1.0 mM IPTG, after an incubation of 6 h at 37 degrees C. Under these conditions, the recombinant strain K protease was, subsequently, released into the periplasm of E. coli BL21 (DE3) with an optimum proteolytic activity detected at 1.0112 U/ml. To date, this is the first reported expression of alkaline protease (aprA) with such remarkable property in Escherichia coli. Wiley-VCH Verlag GmbH & Co. KGaA 2010 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/15709/1/15709.pdf Raja Abdul Rahman, Raja Noor Zaliha and Lee, Poh Geok and Wong, Chee Fah and Basri, Mahiran and Salleh, Abu Bakar (2010) Molecular investigation of a gene encoding organic solvent-tolerant alkaline protease from Pseudomonas aeruginosa strain K. Journal of Basic Microbiology, 50 (2). pp. 143-149. ISSN 0233-111X; ESSN: 1521-4028 http://onlinelibrary.wiley.com/wol1/doi/10.1002/jobm.200900133/abstract 10.1002/jobm.200900133 |
| spellingShingle | Raja Abdul Rahman, Raja Noor Zaliha Lee, Poh Geok Wong, Chee Fah Basri, Mahiran Salleh, Abu Bakar Molecular investigation of a gene encoding organic solvent-tolerant alkaline protease from Pseudomonas aeruginosa strain K |
| title | Molecular investigation of a gene encoding organic solvent-tolerant alkaline protease from Pseudomonas aeruginosa strain K |
| title_full | Molecular investigation of a gene encoding organic solvent-tolerant alkaline protease from Pseudomonas aeruginosa strain K |
| title_fullStr | Molecular investigation of a gene encoding organic solvent-tolerant alkaline protease from Pseudomonas aeruginosa strain K |
| title_full_unstemmed | Molecular investigation of a gene encoding organic solvent-tolerant alkaline protease from Pseudomonas aeruginosa strain K |
| title_short | Molecular investigation of a gene encoding organic solvent-tolerant alkaline protease from Pseudomonas aeruginosa strain K |
| title_sort | molecular investigation of a gene encoding organic solvent tolerant alkaline protease from pseudomonas aeruginosa strain k |
| url | http://psasir.upm.edu.my/id/eprint/15709/1/15709.pdf |
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