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KMID : 1094720100150020273
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Biotechnology and Bioprocess Engineering
2010 Volume.15 No. 2 p.273 ~ p.276
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Effect of Growth Temperature, Induction, and Molecular Chaperones on the Solubilization of Over-expressed Cellobiose Phosphorylase from Cellvibrio Gilvus under in vivo Conditions
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Singh S. P.
Purohit M. K.
Aoyagi C.
Kitaoka M.
Hayashi K.
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Abstract
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In vivo folding of many proteins can be facilitated by growth temperature, extent of induction, and molecular chaperones, which prevent over-expressed protein from being trapped into insoluble inclusion bodies. In the present report, we describe the role of molecular chaperones and growth temperature on the solubilization of overexpressed Cellobiose Phosphorylase (CBP) in Escherichia coli. The growth of host at low temperature enhanced enzyme in soluble fraction. Similarly, induction of target gene at low level of IPTG also yielded higher enzyme in soluble fraction. The synergistic effect of low temperature and induction on the prevention of inclusion bodies was also evident from our results. In addition, co-expression of the target gene with two types of molecular chaperones (GroESL and KODHsp) was also attempted. However, none of these chaperones enhanced the solubilization under in vivo conditions. Nevertheless, effective role of low growth temperature coupled with low level of induction appeared to be an attractive feature for producing recombinant protein.
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KEYWORD
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over-expression, solubilization, molecularchaperone, cellobiose phosphorylase, Cellvibrio gilvus
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