Abstract
Yeast’s extracellular expression provides a cost-efficient means of producing recombinant proteins of academic or commercial interests. However, depending on the protein to be expressed, the production occasionally results in a poor yield, which is frequently accompanied with a deteriorated growth of the host. Here we describe our simple approach, high cell-density expression, to circumvent the cellular toxicity and achieve in a production of a certain range of “difficult-to-express” secretory protein in preparative amount. The system features an ease of performing: (1) precultivate yeast cells to the stationary phase in non-inducing condition, (2) suspend the cells to a small aliquot of inducing medium to form a high cell-density suspension or “a phalanx,” and then (3) give a sufficient aeration to the phalanx. Factors and pitfalls that affect the system’s performance are also described.
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Acknowledgment
This work was financially supported by Industrial Technology Research Grant Program (04A01042) from New Energy and Industrial Technology Development Organization (NEDO) of Japan and partly by Grant-in-Aid for Young Scientist (21760645) from Ministry of Education, Culture, Sports and Technology (MEXT). The cDNA for L. edodes Lcc4 was a kind gift from Drs. Yuko Nakagawa and Akira Yano (Iwate Biotechnology Research Center). We are grateful to Mr. Koichi Kimata and Yuta Saito (University of Shizuoka) for their technical supports. Suggestions and advices on fermentation engineering by Prof. Tsuneo Yamane are also appreciated.
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Kawarasaki, Y., Kurose, T., Ito, K. (2015). High Cell-Density Expression System: Yeast Cells in a Phalanx Efficiently Produce a Certain Range of “Difficult-to-Express” Secretory Recombinant Proteins. In: García-Fruitós, E. (eds) Insoluble Proteins. Methods in Molecular Biology, vol 1258. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2205-5_9
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DOI: https://doi.org/10.1007/978-1-4939-2205-5_9
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