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Heterologous expression of cholesterol oxidase in Bifidobacterium longum under the control of 16S rRNA gene promoter of bifidobacteria

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Abstract

We have constructed a constitutive high-level-expression vector for the genus Bifidobacterium and used it to express cholesterol oxidase from Streptomyces coelicola. The promoter region of the 16S rRNA gene was amplified by inverse PCR and used for the construction of pBES16PR. The optimal ribosome-binding site (RBS) for Bifidobacterium was incorporated in pBES16PR. In order to test the efficacy of this expression vector, we constructed pBES16PR-CHOL with the structural gene for cholesterol oxidase under the control of the 16S rRNA promoter, and used it to transform Bifidobacterium longum. The gene was successfully expressed and high level of cholesterol oxidase activity was obtained in B. longum. This is the first report of an expression vector for the genus Bifidobacterium using a 16S rRNA gene promoter and successful expression of cholesterol oxidase.

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Acknowledgments

This work was supported by the 21C Frontier Microbial Genomics and Applications Center Program, Ministry of Science and Technology (Grant MG05-0307-1-0), Republic of Korea.

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Authors and Affiliations

  1. Research center, BIFIDO Co., Ltd., Seoul, 151-818, Korea

    Myeong Soo Park, Bin Kwon & Geun Eog Ji

  2. Department of Nutrition and Hotel Culinary Art, Anyang Technical College, Anyang, Kyeonggi, Korea

    Myeong Soo Park

  3. Department of Food and Nutrition, Seoul National University, San 56-1, Shillim-dong, Kwanak-gu, Seoul, 151-742, South Korea

    Bin Kwon & Geun Eog Ji

  4. Research center, Korean Yakult, Kyeonggi, Korea

    Jae Jung Shim & Chul Sung Huh

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  1. Myeong Soo Park
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  2. Bin Kwon
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  3. Jae Jung Shim
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  4. Chul Sung Huh
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  5. Geun Eog Ji
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Correspondence to Geun Eog Ji.

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Myeong Soo Park and Bin Kwon contributed equally.

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Park, M.S., Kwon, B., Shim, J.J. et al. Heterologous expression of cholesterol oxidase in Bifidobacterium longum under the control of 16S rRNA gene promoter of bifidobacteria. Biotechnol Lett 30, 165–172 (2008). https://doi.org/10.1007/s10529-007-9514-3

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  • DOI: https://doi.org/10.1007/s10529-007-9514-3

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