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.
Department of Applied Chemistry, Kobe City College of Technology, Gakuen-higashimachi, Nishi-ku, Kobe 651-2194, Japan.
Seiji
S
Shibasaki
Atsuo
A
Tanaka
Mitsuyoshi
M
Ueda
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