Development of combinatorial bioengineering using yeast cell surface display--order-made design of cell and protein for bio-monitoring.
A genetic system to display proteins as their active and functional forms on the cell surface of yeast, Saccharomyces cerevisiae, has been exploited. Surface-engineered (arming) cells displaying amylase or cellulase could assimilate starch or cellulose as the sole carbon source, although S. cerevisiae can not intrinsically assimilate them. Arming cells with a green fluorescent protein (GFP) from Aequorea victoria can emit green fluorescence from the cell surface in response to the environmental conditions. From these results, we attempted to construct a system to monitor the foreign protein production in yeast by simultaneous displaying the enhanced GFP (EGFP). The expression in yeast of the Escherichia coli beta-galactosidase-encoding gene was examined as an example of intracellular production and that of the human interferon-alpha (omega, IFN-omega)-encoding gene as an example of extracellular production. Their productions and the simultaneous surface-display of EGFP as a reporter were controlled by the same promoter, GAL1. The relationship among fluorescence signals and their productions was evaluated. The surface-display system, unlike one using tag-proteins, would be able to facilitate the monitoring of native protein productions in bioprocesses using living cells in real time by the combination of promoters and GFP variants.
Shibasaki
Seiji
S
Department of Applied Chemistry, Kobe City College of Technology, Gakuen-higashimachi, Nishi-ku, Kobe 651-2194, Japan.
Tanaka
Atsuo
A
Ueda
Mitsuyoshi
M
D001698:Biomedical Engineering / Q000379:methods; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D020650:Combinatorial Chemistry Techniques; D005690:Galactose / Q000032:analysis* / Q000378:metabolism*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D016898:Interferon-alpha / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000378:metabolism; D015203:Reproducibility of Results; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis* / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*
D016898:Interferon-alpha; D008164:Luminescent Proteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D049452:Green Fluorescent Proteins; D005690:Galactose