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KMID : 1094720060110050414
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Biotechnology and Bioprocess Engineering
2006 Volume.11 No. 5 p.414 ~ p.419
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Effect of molecular chaperones on the soluble expression of alginate lyase in E. coli
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Shin Eun-Jung
Park So-Lim
Jeon Seong-Soo
Lee Jin-Woo
Kim Young-Tae
Kim Yeon-Hee
Nam Soo-Wan
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Abstract
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When the alginate lyase gene (aly) fromPseudoalteromonas elyakovii was expressed inE. coli, most of the gene product was organized as aggregated insoluble particles known as inclusion bodies. To examine the effects of chaperones on soluble and nonaggregated form of alginate lyase inE. coli, we constructed plasmids designed to permit the coexpression ofaly and the DnaK/DnaJ/GrpE or GroEL/ES chaperones. The results indicate that coexpression ofaly with the Dnak/DnaJ/GrpE chaperone together had a marked effect on the yield alginate lyase as a soluble and active form of the enzyme. It is speculated this result occurs through facilitation of the correct folding of the protein. The optimal concentration ofl-arabinose required for the induction of the DnaK/DnaJ/GrpE chaperone was found to be 0.05 mg/mL. An analysis of the protein bands on SDS-PAGE gel indicated that at least 37% of total alginate lyase was produced in the soluble fraction when the DnaK/DnaJ/GrpE chaperone was coexpressed.
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KEYWORD
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alginate lyase, molecular chaperone, coexpression, E. coli
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