Characterization and solvent stability of strep-tagged purified recombinant lipase from thermostable and solvent tolerant Bacillus sp. strain 42.

A 1.2 kb lipase gene (AY 78735) from solvent stable and thermostable Bacillus sp. strain 42 was overexpressed in a heterologous system that allowed for an extensive characterization of its solvent stability and thermostability. An overexpression was achieved using pET51b vector with Escherichia col...

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Main Authors:	Tengku Abdul Hamid, Tengku Haziyamin, Mohamed , Abdallah Eltaweel, Raja Abdul Rahman, Raja Noor Zaliha, Basri, Mahiran, Salleh, Abu Bakar
Format:	Article
Language:	English
Published:	Springer 2009
Subjects:	Q Science (General)
Online Access:	http://irep.iium.edu.my/1558/ http://irep.iium.edu.my/1558/ http://irep.iium.edu.my/1558/1/Tg_Annal-Micribiology.pdf

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recordtype	eprints
spelling	iium-15582011-09-06T06:20:40Z http://irep.iium.edu.my/1558/ Characterization and solvent stability of strep-tagged purified recombinant lipase from thermostable and solvent tolerant Bacillus sp. strain 42. Tengku Abdul Hamid, Tengku Haziyamin Mohamed , Abdallah Eltaweel Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Salleh, Abu Bakar Q Science (General) A 1.2 kb lipase gene (AY 78735) from solvent stable and thermostable Bacillus sp. strain 42 was overexpressed in a heterologous system that allowed for an extensive characterization of its solvent stability and thermostability. An overexpression was achieved using pET51b vector with Escherichia coli host strain BL21(DE3)pLysS, in which optimum expression was at 22-24 h incubation at 37 °C, with lipase activity reached at 80.0 U mL-1 (specific activity 160.0 U mg-1), after induction by 0.5 mM IPTG. This expression was 11.5 fold higher and superseded the pQE-30UA/M15(pREP4) host-vector system, which only achieved at 17.0 U mL-1 (34.0 U mg-1). The fusion lipase contains N-terminal Strep-tag II affinity tag that in one step of purification, the lipase was purified to homogeneity using Strep-tag II agarose column. The lipase was purified at 1.3 fold and 70% recovery with the elution fraction gave a band of 43 kDa in SDS-PAGE. The purified fusion lipase was most active at 70 °C and pH 8.0, and was stable in a broad pH range of 7-10. It showed hydrolysis preference towards olive, sunflower and corn oils. Based on solvent stability studies in 30 min pre-incubation in 25% v/v solvents with a shaking rate at 150 strokes per min, the purified Lip 42 showed a different residual activity profiles depending on solvents and temperatures. Lip 42 was found be stable in polar organic solvents such as DMSO, DMF, acetone, methanol, heptanol and octanol, which could make it as a potential biocatalyst for the use in industrial biodiesel production. Springer 2009 Article PeerReviewed application/pdf en http://irep.iium.edu.my/1558/1/Tg_Annal-Micribiology.pdf Tengku Abdul Hamid, Tengku Haziyamin and Mohamed , Abdallah Eltaweel and Raja Abdul Rahman, Raja Noor Zaliha and Basri, Mahiran and Salleh, Abu Bakar (2009) Characterization and solvent stability of strep-tagged purified recombinant lipase from thermostable and solvent tolerant Bacillus sp. strain 42. Annals of Microbiology, 59 (1). pp. 111-118. ISSN 1590-4261 http://www.springerlink.com/content/r69471q47k057653/
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English
topic	Q Science (General)
spellingShingle	Q Science (General) Tengku Abdul Hamid, Tengku Haziyamin Mohamed , Abdallah Eltaweel Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Salleh, Abu Bakar Characterization and solvent stability of strep-tagged purified recombinant lipase from thermostable and solvent tolerant Bacillus sp. strain 42.
description	A 1.2 kb lipase gene (AY 78735) from solvent stable and thermostable Bacillus sp. strain 42 was overexpressed in a heterologous system that allowed for an extensive characterization of its solvent stability and thermostability. An overexpression was achieved using pET51b vector with Escherichia coli host strain BL21(DE3)pLysS, in which optimum expression was at 22-24 h incubation at 37 °C, with lipase activity reached at 80.0 U mL-1 (specific activity 160.0 U mg-1), after induction by 0.5 mM IPTG. This expression was 11.5 fold higher and superseded the pQE-30UA/M15(pREP4) host-vector system, which only achieved at 17.0 U mL-1 (34.0 U mg-1). The fusion lipase contains N-terminal Strep-tag II affinity tag that in one step of purification, the lipase was purified to homogeneity using Strep-tag II agarose column. The lipase was purified at 1.3 fold and 70% recovery with the elution fraction gave a band of 43 kDa in SDS-PAGE. The purified fusion lipase was most active at 70 °C and pH 8.0, and was stable in a broad pH range of 7-10. It showed hydrolysis preference towards olive, sunflower and corn oils. Based on solvent stability studies in 30 min pre-incubation in 25% v/v solvents with a shaking rate at 150 strokes per min, the purified Lip 42 showed a different residual activity profiles depending on solvents and temperatures. Lip 42 was found be stable in polar organic solvents such as DMSO, DMF, acetone, methanol, heptanol and octanol, which could make it as a potential biocatalyst for the use in industrial biodiesel production.
format	Article
author	Tengku Abdul Hamid, Tengku Haziyamin Mohamed , Abdallah Eltaweel Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Salleh, Abu Bakar
author_facet	Tengku Abdul Hamid, Tengku Haziyamin Mohamed , Abdallah Eltaweel Raja Abdul Rahman, Raja Noor Zaliha Basri, Mahiran Salleh, Abu Bakar
author_sort	Tengku Abdul Hamid, Tengku Haziyamin
title	Characterization and solvent stability of strep-tagged purified recombinant lipase from thermostable and solvent tolerant Bacillus sp. strain 42.
title_short	Characterization and solvent stability of strep-tagged purified recombinant lipase from thermostable and solvent tolerant Bacillus sp. strain 42.
title_full	Characterization and solvent stability of strep-tagged purified recombinant lipase from thermostable and solvent tolerant Bacillus sp. strain 42.
title_fullStr	Characterization and solvent stability of strep-tagged purified recombinant lipase from thermostable and solvent tolerant Bacillus sp. strain 42.
title_full_unstemmed	Characterization and solvent stability of strep-tagged purified recombinant lipase from thermostable and solvent tolerant Bacillus sp. strain 42.
title_sort	characterization and solvent stability of strep-tagged purified recombinant lipase from thermostable and solvent tolerant bacillus sp. strain 42.
publisher	Springer
publishDate	2009
url	http://irep.iium.edu.my/1558/ http://irep.iium.edu.my/1558/ http://irep.iium.edu.my/1558/1/Tg_Annal-Micribiology.pdf
first_indexed	2023-09-18T20:08:55Z
last_indexed	2023-09-18T20:08:55Z
_version_	1777407346294652928

Characterization and solvent stability of strep-tagged purified recombinant lipase from thermostable and solvent tolerant Bacillus sp. strain 42.

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