title: High cell-density expression system: yeast cells in a phalanx efficiently produce a certain range of "difficult-to-express" secretory recombinant proteins.
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.
authors.affiliation: Biomolecular Engineering Laboratory, Graduate School of Integrated Pharmaceutical and Nutritional Sciences, University of Shizuoka, Yada 52-1, Suruga-ku, Shizuoka, 422-8526, Japan, kawarsky@u-shizuoka-ken.ac.jp.
authors.forename: Yasuaki
authors.initials: Y
authors.lastname: Kawarasaki
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kurose
authors.forename: Keisuke
authors.initials: K
authors.lastname: Ito
mesh_terms: D000818:Animals; D002452:Cell Count / Q000379:methods; D015870:Gene Expression / Q000235:genetics*; D006801:Humans; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism*; D015003:Yeasts / Q000235:genetics* / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins