In situ intein-mediated multiprotein assembly via engineered cross-species consortia

TL;DR

Scientists created a system where microbes work together to assemble proteins in real time, without needing purification steps.

Contribution

A programmable microbial consortia platform for in situ protein splicing using split inteins and synchronized lysis.

Findings

Engineered E. coli and P. pastoris work together to assemble proteins extracellularly in culture.

The system uses quorum-sensing and eukaryotic secretion to enable scalable, one-pot protein synthesis.

The platform supports logic computation and antibiotic resistance engineering for adaptive biomanufacturing.

Abstract

Inteins can connect flanking external proteins into a new protein fragment and excise themselves. Here, we report the in situ splicing of proteins by engineered microbial consortia. This study pioneers a programmable microbial consortia platform enabling in situ protein splicing through split intein-mediated assembly. Engineered Escherichia coli with the ePop autolysis system release intein-fused protein fragments via synchronized lysis, while Pichia pastoris secretes complementary domains, enabling extracellular reconstitution directly in culture. With the application of integrating quorum-sensing controls and eukaryotic secretion pathways, this approach bypasses in vitro purification, supporting scalable one-pot synthesis of multiple functional proteins. The platform’s versatility in logic computation and antibiotic resistance engineering highlights its potential for adaptive biomanufacturing and context-aware biomaterial design.

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