Biology subjects

Tesoriero, R. F.

Publications and source records attributed to Tesoriero, R. F..

2 recordsLinked to original sources

bioRxiv2026

A Computational Framework for Domain Insertion into Type IV Pili for Bacterial Display and Living Material Assembly

Bacterial surface structures have enabled display systems with broad impact across biotechnology, but their narrow host range limits their deployment into diverse species. Conversely, Type IV Pili (TFP) are ubiquitous, structurally conserved appendages found across bacteria, but have been minimally explored for display. Here, we describe a computational framework for predicting viable insertion sites in major pilins for stable TFP-mediated display, which we apply to the major pilin PilA1 of the cyanobacterium Synechocystis sp. PCC 6803 to enable covalent binding to living materials. By analyzing an Alphafold3-generated PilA1 monomer alongside known multimeric TFP multimers, our pipeline identifies non-interfacial, solvent-accessible, and flexible sites for optimal PilA1 display. We probe these sites with both full-length and truncated SpyCatcher003 at two different expression levels. We show that cells expressing these PilA1-SpyCatcher003 fusions maintain up to 8-fold higher levels of cell suspension than previous C-terminal PilA1 display platforms, suggesting improved TFP assembly despite more than a two-fold increase in cargo size. Additionally, we validate SpyCatcher003 reactivity across the engineered strains, enabling covalent attachment of SpyTag003-containing proteins on the Synechocystis surface. Lastly, we utilize this covalent patterning to achieve a four-fold increase in Synechocystis loading into a living material without compromising its viscoelastic or mechanical properties. Taken together, this work provides a predictive framework for TFP engineering, and opens the door towards programmable surface display across the breadth of bacterial species.

synthetic biology↗

bioRxiv2021

A de novo matrix for macroscopic living materials from bacteria

Summary ParagraphEngineered living materials (ELMs) are composites of living cells embedded in a biopolymer matrix that combine the desirable properties of natural biomaterials with non-natural, tailored properties. ELMs with a wide range of sophisticated biological functions have been created by engineering the embedded cells using synthetic biology. Engineering a de novo biomolecular matrix would offer control over material assembly, structure, and composition, thus enabling us to grow macroscopic ELMs with customizable mechanical properties. However, we have lacked the genetic tools and design rules to genetically encode a synthetic matrix that programs collective cell self-organization into macroscopic structures. Here we report growth of macroscopic ELMs from Caulobacter crescentus cells that display and secrete an engineered self-interacting protein. This protein formed an extracellular de novo matrix and assembled cells into hierarchically-ordered, centimeter-scale ELMs. We showed that the mechanical, catalytic, and morphological properties of these ELMs can be tuned through genetic modification of the matrix. Our work identifies novel genetic tools, design and assembly rules for growing macroscopic ELMs with both wide-ranging mechanical properties and customizable functions. We anticipate the modularity of this approach will permit the incorporation of different protein polymers in the de novo matrix, thus allowing to generate ELMs with a variety of desired structures and compositions of the bulk material. We envision specific matrix properties that can be combined synergistically with existing cellular functions to greatly expand the opportunities for ELMs in human health, energy, and the environment.

synthetic biology↗