@article{01K2YAH5ACN0J2PXNFMQ0JKVAG,
  abstract     = {{Designer cellulosomes (DCs) are precisely engineered multi-enzyme complexes aimed at lignocellulose saccharification. Achieving spontaneous designer cellulosome display on yeast cell surfaces has been a long-term objective to enhance consolidated bioprocesses with concurrent ethanol production. A “self-assembly” approach involves simultaneous scaffoldin display and docking enzyme secretion. However, challenges arise from the size and complexity of designer cellulosomes, coupled with the yeast cells’ limited capacity for heterologous protein expression. A comprehensive examination of Saccharomyces cerevisiae as a host for DC expression remains unaddressed. We meticulously examined the capability of S. cerevisiae to produce and display a fluorescent protein complex, which mimics the designer cellulosome architecture and allows for convenient detection of all individual components on the cell surface, using flow cytometry and confocal microscopy. Population-wide analysis revealed a fluorescent protein complex production efficiency of approximately 10%. Single-cell analysis highlighted a clear mutual influence between the expression of scaffoldin and docking proteins, impacting cellular fitness. Newly emerging buds were identified as hotspots for scaffoldin display. The finite capacity of yeast cells to produce heterologous proteins was identified as a major bottleneck. While distributing the cellular load among multiple hosts within a synthetic yeast consortium can alleviate this burden, the use of fluorescent protein complex surface display has visualized the heterogeneity and constraints of S. cerevisiae as a host for designer cellulosome expression at the population and single cell level. This study provides a realistic assessment of the challenges in achieving efficient S. cerevisiae-based DC display for consolidated bioprocessing.}},
  articleno    = {{e00750-25}},
  author       = {{Lamote, Babette and Cremelie, Emma and Demeyere, Kristel and De Groote, Philippe and Grimon, Dennis and Vanderstraeten, Julie and Meyer, Evelyne and De Mey, Marjan and Briers, Yves}},
  issn         = {{2165-0497}},
  journal      = {{MICROBIOLOGY SPECTRUM}},
  keywords     = {{designer cellulosomes,Saccharomyces cerevisiae,yeast surface display,flow cytometry,confocal microscopy,fluorescent proteins,SACCHAROMYCES-CEREVISIAE,ETHANOL,SACCHARIFICATION,MINICELLULOSOME,NANOMACHINES,SECRETION}},
  language     = {{eng}},
  number       = {{9}},
  pages        = {{16}},
  title        = {{Single-cell analysis of yeast surface display for designer cellulosome applications using a fluorescent protein complex}},
  url          = {{http://doi.org/10.1128/spectrum.00750-25}},
  volume       = {{13}},
  year         = {{2025}},
}

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