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Effect of molecular chaperones on the soluble expression of alginate lyase inE. coli

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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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Authors and Affiliations

  1. Department of Biomaterial Control, Dong-Eui University, 614-714, Busan, Korea

    Eun-Jung Shin, Sung-Jong Jeon & Soo-Wan Nam

  2. BioLeaders Corp., 301-212, Daejeon, Korea

    So-Lim Park

  3. Department of Biotechnology and Bioengineering, Dong-Eui University, 614-714, Busan, Korea

    Sung-Jong Jeon & Soo-Wan Nam

  4. Department of Biotechnology, Dong-A University, 604-714, Busan, Korea

    Jin-Woo Lee

  5. Department of Microbiology, Pukyong National University, 608-737, Busan, Korea

    Young-Tae Kim

  6. Department of Biotechnology, Osaka University, 565-0871, Osaka, Japan

    Yeon-Hee Kim

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  1. Eun-Jung Shin
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Shin, EJ., Park, SL., Jeon, SJ. et al. Effect of molecular chaperones on the soluble expression of alginate lyase inE. coli . Biotechnol. Bioprocess Eng. 11, 414–419 (2006). https://doi.org/10.1007/BF02932308

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  • DOI: https://doi.org/10.1007/BF02932308

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