Cell-Free Synthetic Glycobiology: Designing and Engineering Glycomolecules Outside of Living Cells

Thapakorn Jaroentomeechai, May N. Taw, Mingji Li, Alicia Aquino, Ninad Agashe, Sean Chung, Michael C. Jewett, Matthew P. DeLisa*

*Corresponding author for this work

Research output: Contribution to journalReview articlepeer-review

26 Scopus citations

Abstract

Glycans and glycosylated biomolecules are directly involved in almost every biological process as well as the etiology of most major diseases. Hence, glycoscience knowledge is essential to efforts aimed at addressing fundamental challenges in understanding and improving human health, protecting the environment and enhancing energy security, and developing renewable and sustainable resources that can serve as the source of next-generation materials. While much progress has been made, there remains an urgent need for new tools that can overexpress structurally uniform glycans and glycoconjugates in the quantities needed for characterization and that can be used to mechanistically dissect the enzymatic reactions and multi-enzyme assembly lines that promote their construction. To address this technology gap, cell-free synthetic glycobiology has emerged as a simplified and highly modular framework to investigate, prototype, and engineer pathways for glycan biosynthesis and biomolecule glycosylation outside the confines of living cells. From nucleotide sugars to complex glycoproteins, we summarize here recent efforts that harness the power of cell-free approaches to design, build, test, and utilize glyco-enzyme reaction networks that produce desired glycomolecules in a predictable and controllable manner. We also highlight novel cell-free methods for shedding light on poorly understood aspects of diverse glycosylation processes and engineering these processes toward desired outcomes. Taken together, cell-free synthetic glycobiology represents a promising set of tools and techniques for accelerating basic glycoscience research (e.g., deciphering the “glycan code”) and its application (e.g., biomanufacturing high-value glycomolecules on demand).

Original languageEnglish (US)
Article number645
JournalFrontiers in Chemistry
Volume8
DOIs
StatePublished - Jul 29 2020

Funding

Funding. This work was supported by the Defense Threat Reduction Agency (HDTRA1-15-10052 and HDTRA1-20-10004 to MD and MJ), National Institutes of Health Grant (R01 GM137314-01 and R01 GM127578-01 to MD and 1U19AI142780-01 to MJ), National Science Foundation (Grants # CBET 1159581, CBET 1264701, and CMMI 1728049 to MD; MCB 1716766 to MJ; and CBET 1936823 and MCB 1413563 to MD and MJ), the Bill and Melinda Gates Foundation (Grant OPP1217652 to MD and MJ), the DARPA 1000 Molecules Program (HR0011-15-C-0084), the Human Frontiers Science Program (Grant RGP0015/2017), the David and Lucile Packard Foundation (to MJ), and the Camille Dreyfus Teacher-Scholar Program (to MJ). TJ was supported by a Royal Thai Government Fellowship and the Cornell Fleming Graduate Scholarship. AA was supported by an NIH Chemical Biology Interface (CBI) training fellowship (supporting grant T32GM008500) and a National Science Foundation Graduate Research Fellowship (DGE-1650441).

Keywords

  • Cell-free system
  • chemoenzymatic synthesis
  • glycoprotein therapeutics and vaccines
  • glycoscience
  • metabolic glycoengineering
  • nucleotide sugars
  • post-translational modification
  • synthetic biology

ASJC Scopus subject areas

  • General Chemistry

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