Effect of molecular chaperones on the soluble expression of alginate lyase inE. coli |
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Authors: | Eun-Jung Shin So-Lim Park Sung-Jong Jeon Jin-Woo Lee Young-Tae Kim Yeon-Hee Kim Soo-Wan Nam |
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Institution: | (1) Department of Chemical Engineering, Kwangwoon University, 447-1 Wolgye-dong, Nowon-gu, Seoul, 139-701, Republic of Korea; |
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Abstract: | 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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