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351.
Gokce Altin-Yavuzarslan Sierra M. Brooks Shuo-Fu Yuan James O. Park Hal S. Alper Alshakim Nelson 《Advanced functional materials》2023,33(24):2300332
Engineered living materials (ELMs) combine living cells with polymeric matrices to yield unique materials with programmable functions. While the cellular platform and the polymer network determine the material properties and applications, there are still gaps in the ability to seamlessly integrate the biotic (cellular) and abiotic (polymer) components into singular materials, then assemble them into devices and machines. Herein, the additive-manufacturing of ELMs wherein bioproduction of metabolites from the encapsulated cells enhanced the properties of the surrounding matrix is demonstrated. First, aqueous resins are developed comprising bovine serum albumin (BSA) and poly(ethylene glycol diacrylate) (PEGDA) with engineered microbes for vat photopolymerization to create objects with a wide array of 3D form factors. The BSA-PEGDA matrix afforded hydrogels that are mechanically stiff and tough for use in load-bearing applications. Second, the continuous in situ production of l -DOPA, naringenin, and betaxanthins from the engineered cells encapsulated within the BSA-PEGDA matrix is demonstrated. These microbial metabolites bioaugmented the properties of the BSA-PEGDA matrix by enhancing the stiffness (l -DOPA) or resistance to enzymatic degradation (betaxanthin). Finally, the assembly of the 3D printed ELM components into mechanically functional bolts and gears to showcase the potential to create functional ELMs for synthetic living machines is demonstrated. 相似文献
352.
Sierra J. Williams Jordan A. Gewing-Mullins Whitney K. Lieberman Bethany Kolbaba-Kartchner Reema Iqbal Hana M. Burgess Clair M. Colee Marya Y. Ornelas Edison S. Reid-McLaughlin Prof. Dr. Jeremy H. Mills Prof. Dr. Jennifer A. Prescher Prof. Dr. Aaron M. Leconte 《Chembiochem : a European journal of chemical biology》2023,24(6):e202200726
Engineered luciferase-luciferin pairs have expanded the number of cellular targets that can be visualized in tandem. While light production relies on selective processing of synthetic luciferins by mutant luciferases, little is known about the origin of selectivity. The development of new and improved pairs requires a better understanding of the structure−function relationship of bioluminescent probes. In this work, we report a biochemical approach to assessing and optimizing two popular bioluminescent pairs: Cashew/d -luc and Pecan/4′-BrLuc. Single mutants derived from Cashew and Pecan revealed key residues for selectivity and thermal stability. Stability was further improved through a rational addition of beneficial residues. In addition to providing increased stability, the known stabilizing mutations surprisingly also improved selectivity. The resultant improved pair of luciferases are >100-fold selective for their respective substrates and highly thermally stable. Collectively, this work highlights the importance of mechanistic insight for improving bioluminescent pairs and provides significantly improved Cashew and Pecan enzymes which should be immediately suitable for multicomponent imaging applications. 相似文献