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Abstract:
Centralized facilities for genetic engineering, or
“biofoundries”, offer the potential to design organisms to address
emerging needs in medicine, agriculture, industry, and defense.
The field has seen rapid advances in technology, but it is difficult
to gauge current capabilities or identify gaps across projects.
To this end, our foundry was assessed via a timed “pressure test”,
in which 3 months were given to build organisms to produce 10
molecules unknown to us in advance. By applying a diversity of
new approaches, we produced the desired molecule or a closely
related one for six out of 10 targets during the performance
period and made advances toward production of the others as
well. Specifically, we increased the titers of 1-hexadecanol, pyrrolnitrin,
and pacidamycin D, found novel routes to the enediyne
warhead underlying powerful antimicrobials, established a cellfree
system for monoterpene production, produced an intermediate toward vincristine biosynthesis, and encoded 7802 individually
retrievable pathways to 540 bisindoles in a DNA pool. Pathways to tetrahydrofuran and barbamide were designed and
constructed, but toxicity or analytical tools inhibited further progress. In sum, we constructed 1.2 Mb DNA, built 215 strains
spanning five species (Saccharomyces cerevisiae, Escherichia coli, Streptomyces albidof lavus, Streptomyces coelicolor, and Streptomyces
albovinaceus), established two cell-free systems, and performed 690 assays developed in-house for the molecules.
