Recombinant protein condensation inside E. coli enables the development of building blocks for bioinspired materials engineering - Biomimetic spider silk protein as a case study.

ABSTRACT: Recombinant expression of proteins destined to form biological materials often results in poor production yields or loss of their function due to premature aggregation. Recently, liquid-liquid phase separation has been proposed as a mechanism to control protein solubility during expression and accumulation in the cytoplasm. Here, we investigate this process in vivo during the recombinant overexpression of the mimetic spider silk mini-spidroin NT2RepCT in Escherichia coli. The protein forms intracellular liquid-like condensates that shift to a solid-like state triggered by a decrease in their microenvironmental pH. These features are also maintained in the purified sample in vitro both in the presence of a molecular crowding agent mimicking the bacterial intracellular e

SUBMITTER: Gabryelczyk B 

PROVIDER: S-EPMC9677211 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature