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Molecular cloning and expression ofThiobacillus ferrooxidans chromosomal ribulose bisphosphate carboxylase genes inEscherichia coli

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Abstract

The genes coding ford-ribulose-1,5-bisphosphate carboxylase (RuBPCase) from an iron-oxidizing bacterium,Thiobacillus ferrooxidans, were cloned into anEscherichia coli plasmid, pUC18. The recombinant plasmid, termed pTR11, contained a 4.0-kb PstI fragment including the entire coding regions for both large and small subunits of RuBPCase.Escherichia coli carrying pTR11 did not show any CO2-fixing activity. However, a derivative plasmid with an appropriate deletion, which was placed under the control of atac promoter, conferred ribulose bisphosphate-dependent CO2-fixing activity on the host cell. Analysis of gel-filtration chromatography of the RuBPCase synthesized inE. coli revealed that it had a hexadecameric form like the native enzyme ofT. ferrooxidans.

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Authors and Affiliations

  1. Research Center, Dowa Mining Co. Ltd., Hachioji, Tokyo, Japan

    Chihiro Inoue

  2. Laboratory of Plant Genetic Engineering, Biotechnology Institute, Akita Prefectural College of Agriculture, Ohgata, 010-04, Akita, Japan

    Tomonobu Kusano, Kazuyuki Sugawara, Chihiro Inoue & Nobuhiro Suzuki

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  1. Tomonobu Kusano
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  2. Kazuyuki Sugawara
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  3. Chihiro Inoue
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  4. Nobuhiro Suzuki
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Kusano, T., Sugawara, K., Inoue, C. et al. Molecular cloning and expression ofThiobacillus ferrooxidans chromosomal ribulose bisphosphate carboxylase genes inEscherichia coli . Current Microbiology 22, 35–41 (1991). https://doi.org/10.1007/BF02106210

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