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Indian Journal of Biochemistry and Biophysics (IJBB) >
IJBB Vol.48 [2011] >
IJBB Vol.48(6) [December 2011] >
| Title: | Functional fusion expression of sunflower multicystatin in E. coli and its comparison with a single domain cystatin |
| Authors: | Gholizadeh, Ashraf
Kohnehrouz, B Baghban |
| Keywords: | Cystatin
Expression
Fusion protein
Inhibitor
Sunflower multicystatin
Phytocystatin |
| Issue Date: | Dec-2011 |
| Publisher: | NISCAIR-CSIR, India |
| Abstract: | Identification of the molecular structure and novel biophysiological
functions of plant cystatins or phytocystatins is of great interest in the
field of molecular biology. The important
requirements for these are the efficient production, purification and
correctly folded forms of these proteins. We report here the cloning, easy
expression and characterization of a sunflower multicystatin (SMC) as a
functional fusion protein in E. coli. For the first time, the amplified
cystatin coding region was expressed as a part of maltose-binding fusion
protein using pMALc2X over-expression vector in TB1 strain of
E. coli without affecting the recombinant bacterial growth. In
comparison to the previously prepared recombinant SMC (rSMC), a high amount
(~44 mg/L of bacterial cell culture) of purified fused SMC (fSMC) was obtained
using single-step purification method. fSMC strongly inhibited papain activity in
vitro as compared to Celosia single-domain cystatin. Purified
fSMC may be used for basic biochemical, pharmacological or clinical studies
without the cleavage of its fusion parts. |
| Page(s): | 375-379 |
| CC License: |  CC Attribution-Noncommercial-No Derivative Works 2.5 India |
| ISSN: | 0975-0959 (Online); 0301-1208 (Print) |
| Source: | IJBB Vol.48(6) [December 2011]
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