Growth-coupled microbial biosynthesis of the animal pigment xanthommatin

Bushin, L. B.; Alter, T. B.; Alvan-Vargas, M. V. G.; Dürr, L.; Olson, E. C.; Avila, M. J.; Puiggene, O.; Kim, T.; Deravi, L. F.; Feist, A. M.; Nikel, P. I.; Moore, B. S.

doi:10.1101/2024.10.04.616593

bioRxiv · 10.1101/2024.10.04.616593

Growth-coupled microbial biosynthesis of the animal pigment xanthommatin

Bushin, L. B.·Alter, T. B.·Alvan-Vargas, M. V. G.·Dürr, L.·Olson, E. C.·Avila, M. J.·Puiggene, O.·Kim, T.·Deravi, L. F.·Feist, A. M.·Nikel, P. I.·Moore, B. S.

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Abstract

The mining of genomes across life has unearthed a bounty of biosynthetic potential to diverse molecules key to a biobased future. While the heterologous expression of metabolic pathways has achieved broad success, most approaches suffer a similar fate in low initial production levels that require extensive, resource-heavy iterative strain engineering refinement. Herein we introduce a growth-coupled biosynthetic (GrowBio) strategy that irrevocably connects microbial growth with specialized compound production. We demonstrate the plug-and-play versatility of GrowBio in the production of the structurally complex animal biopigment xanthommatin, a color-changing ommochrome with material and cosmetic potential. Xanthommatin biosynthesis directly fuels growth of a newly designed Pseudomonas putida 5,10-methylenetetrahydrofolate auxotroph (PUMA). Aided by genome-scale metabolic modeling, PUMA was designed and built to be controlled by endogenous formate co-produced as a coupled biosynthetic byproduct in the multistep conversion of tryptophan to xanthommatin. Adaptive laboratory evolution was utilized to streamline xanthommatins gram-scale bioproduction via growth rate selection, establishing GrowBio as a promising biotechnological approach for establishing and optimizing the microbial production of value-added molecules.

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Bushin, L. B., Alter, T. B., Alvan-Vargas, M. V. G., Dürr, L., Olson, E. C., Avila, M. J., Puiggene, O., Kim, T., Deravi, L. F., Feist, A. M., Nikel, P. I., Moore, B. S.. 2024-10-06. Growth-coupled microbial biosynthesis of the animal pigment xanthommatin. https://doi.org/10.1101/2024.10.04.616593

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