High level expression and characterization of a novel thermostable, organic solvent tolerant, 1,3-regioselective lipase from Geobacillus sp. strain ARM
The mature ARM lipase gene was cloned into the pTrcHis expression vector and over-expressed in Escherichia coli TOP10 host. The optimum lipase expression was obtained after 18h post induction incubation with 1.0mM IPTG, where the lipase activity was approximately 1623-fold higher than wild type. A r...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2011
|
| Online Access: | http://psasir.upm.edu.my/id/eprint/22282/ http://psasir.upm.edu.my/id/eprint/22282/1/High%20level%20expression%20and%20characterization%20of%20a%20novel%20thermostable.pdf |
| _version_ | 1848844443418886144 |
|---|---|
| author | Ebrahimpour, Afshin Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Salleh, Abu Bakar |
| author_facet | Ebrahimpour, Afshin Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Salleh, Abu Bakar |
| author_sort | Ebrahimpour, Afshin |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | The mature ARM lipase gene was cloned into the pTrcHis expression vector and over-expressed in Escherichia coli TOP10 host. The optimum lipase expression was obtained after 18h post induction incubation with 1.0mM IPTG, where the lipase activity was approximately 1623-fold higher than wild type. A rapid, high efficient, one-step purification of the His-tagged recombinant lipase was achieved using immobilized metal affinity chromatography with 63.2% recovery and purification factor of 14.6. The purified lipase was characterized as a high active (7092Umg -1), serine-hydrolase, thermostable, organic solvent tolerant, 1,3-specific lipase with a molecular weight of about 44kDa. The enzyme was a monomer with disulfide bond(s) in its structure, but was not a metalloenzyme. ARM lipase was active in a broad range of temperature and pH with optimum lipolytic activity at pH 8.0 and 65°C. The enzyme retained 50% residual activity at pH 6.0-7.0, 50°C for more than 150min. |
| first_indexed | 2025-11-15T08:31:00Z |
| format | Article |
| id | upm-22282 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T08:31:00Z |
| publishDate | 2011 |
| publisher | Elsevier |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-222822016-09-28T01:11:19Z http://psasir.upm.edu.my/id/eprint/22282/ High level expression and characterization of a novel thermostable, organic solvent tolerant, 1,3-regioselective lipase from Geobacillus sp. strain ARM Ebrahimpour, Afshin Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Salleh, Abu Bakar The mature ARM lipase gene was cloned into the pTrcHis expression vector and over-expressed in Escherichia coli TOP10 host. The optimum lipase expression was obtained after 18h post induction incubation with 1.0mM IPTG, where the lipase activity was approximately 1623-fold higher than wild type. A rapid, high efficient, one-step purification of the His-tagged recombinant lipase was achieved using immobilized metal affinity chromatography with 63.2% recovery and purification factor of 14.6. The purified lipase was characterized as a high active (7092Umg -1), serine-hydrolase, thermostable, organic solvent tolerant, 1,3-specific lipase with a molecular weight of about 44kDa. The enzyme was a monomer with disulfide bond(s) in its structure, but was not a metalloenzyme. ARM lipase was active in a broad range of temperature and pH with optimum lipolytic activity at pH 8.0 and 65°C. The enzyme retained 50% residual activity at pH 6.0-7.0, 50°C for more than 150min. Elsevier 2011-07 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/22282/1/High%20level%20expression%20and%20characterization%20of%20a%20novel%20thermostable.pdf Ebrahimpour, Afshin and Raja Abdul Rahman, Raja Noor Zaliha and Basri, Mahiran and Salleh, Abu Bakar (2011) High level expression and characterization of a novel thermostable, organic solvent tolerant, 1,3-regioselective lipase from Geobacillus sp. strain ARM. Bioresource Technology, 102 (13). pp. 6972-6981. ISSN 0960-8524; ESSN: 1873-2976 http://www.sciencedirect.com/science/article/pii/S0960852411004408 10.1016/j.biortech.2011.03.083 |
| spellingShingle | Ebrahimpour, Afshin Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Salleh, Abu Bakar High level expression and characterization of a novel thermostable, organic solvent tolerant, 1,3-regioselective lipase from Geobacillus sp. strain ARM |
| title | High level expression and characterization of a novel thermostable, organic solvent tolerant, 1,3-regioselective lipase from Geobacillus sp. strain ARM |
| title_full | High level expression and characterization of a novel thermostable, organic solvent tolerant, 1,3-regioselective lipase from Geobacillus sp. strain ARM |
| title_fullStr | High level expression and characterization of a novel thermostable, organic solvent tolerant, 1,3-regioselective lipase from Geobacillus sp. strain ARM |
| title_full_unstemmed | High level expression and characterization of a novel thermostable, organic solvent tolerant, 1,3-regioselective lipase from Geobacillus sp. strain ARM |
| title_short | High level expression and characterization of a novel thermostable, organic solvent tolerant, 1,3-regioselective lipase from Geobacillus sp. strain ARM |
| title_sort | high level expression and characterization of a novel thermostable, organic solvent tolerant, 1,3-regioselective lipase from geobacillus sp. strain arm |
| url | http://psasir.upm.edu.my/id/eprint/22282/ http://psasir.upm.edu.my/id/eprint/22282/ http://psasir.upm.edu.my/id/eprint/22282/ http://psasir.upm.edu.my/id/eprint/22282/1/High%20level%20expression%20and%20characterization%20of%20a%20novel%20thermostable.pdf |