Process Biochemistry, ( ISI ), Volume (48), No (1), Year (2013-10) , Pages (1679-1685)

Title : ( Molecular cloning of a thermo-alkaliphilic lipase from Bacillus subtilis DR8806: Expression and biochemical characterization )

Authors: Shirin Emtenani , Ahmad Asoodeh , Shamsi Emtenani ,

Access to full-text not allowed by authors

Citation: BibTeX | EndNote

A thermo-alkaliphilic lipase from Bacillus subtilis DR8806 was functionally expressed as an N-terminal 6xHis-tagged recombinant enzyme in Escherichia coli BL21 using pET-28a(+) expression vector. Sequence analysis revealed an open reading frame of 639 bp encoding a 212-amino acid protein containing the well-conserved Ala-His-Ser-Met-Gly motif. One-step purification of the His-tagged recombinant lipase was achieved using Ni-NTA affinity chromatography with a specific activity of 1364 U/mg. The purified enzyme with an apparent molecular mass of 26.8 kDa demonstrated the maximum activity at 70 ◦C and pH 8.0 for hydrolysis of p-nitrophenylbutyrate as substrate. The enzyme activity was strongly inhibited by divalent ions of heavy metals such as Hg2+ and Cu2+, while retained over 90% of the original activity in the presence of several reagents including DTNB (5,5-dithiobis-(2-nitrobenzoic acid)), SDS (sodium dodecyl sulfate), urea, DMF (dimethylformamide), DTT (dithiothreitol), glycerol and Triton X-100. While being considerably stable in organic solvents, imidazolium-based ionic liquids (ILs) had stimulatory effects on the activity of purified lipase. Remarkable stabilization of enzyme at alkaline pH and in ionic liquids as well as its thermostability/thermoactivity are among the most fundamental characteristics which offer great potential for various biotechnological applications including detergent formulation, bioremediation processes and biotransformation in non-aqueous media.

Keywords

Cloning Lipase Bacillus subtilis DR8806 Biochemical characterization Ionic
برای دانلود از شناسه و رمز عبور پرتال پویا استفاده کنید.

@article{paperid:1037138,
author = {Emtenani, Shirin and Asoodeh, Ahmad and Emtenani, Shamsi},
title = {Molecular cloning of a thermo-alkaliphilic lipase from Bacillus subtilis DR8806: Expression and biochemical characterization},
journal = {Process Biochemistry},
year = {2013},
volume = {48},
number = {1},
month = {October},
issn = {1359-5113},
pages = {1679--1685},
numpages = {6},
keywords = {Cloning Lipase Bacillus subtilis DR8806 Biochemical characterization Ionic liquids},
}

[Download]

%0 Journal Article
%T Molecular cloning of a thermo-alkaliphilic lipase from Bacillus subtilis DR8806: Expression and biochemical characterization
%A Emtenani, Shirin
%A Asoodeh, Ahmad
%A Emtenani, Shamsi
%J Process Biochemistry
%@ 1359-5113
%D 2013

[Download]