E.i. Dupont De Nemours and Company

Qi Chen

Qiong Cheng

Kristin Ruebling-Jass

Jian Ping Lai

Pierre E Rouviere

Andrew C Eliot

Kevin Michael Croker

Annapurna Sachan

Thomas Eduard Friedmann

Ritu Bhalla

Neeraj Pandey

Methods are provided to obtain recombinant microbial cells having at least one genetic modification that increase the buoyant density of a recombinant microbial cell or the buoyant density of inclusion bodies produced within a recombinant microbial cell. Exemplified are genetic modifications that increase the buoyant density of a recombinant microbial cell expressing heterologous peptides and polypeptides. Increasing expression of the genes ysaB, glyQ, glyS or a combination thereof within the recombinant microbial cell produces cells or inclusion bodies having higher buoyant density. A similar effect was achieved by decreasing or disrupting expression of the endogenous gltA gene. Increases in buoyant density render peptide production more efficient with respect to time and costs.

Recombinant host cells having an increase in buoyant density

Recombinant bacteria having an improved ability to utilize sucrose are provided. These recombinant bacteria have nucleotide sequences encoding sucrose utilization polypeptides integrated into their genome between the yihP gene or its homolog and the yihO gene or its homolog. Additionally, methods of utilizing the recombinant bacteria to produce products such as glycerol and glycerol-derived products are provided.

Recombinant bacteria having improved sucrose utilization

Variant sucrose transporter polypeptides that enable faster sucrose utilization in bacteria are described. Additionally, variant or recombinant bacteria comprising these variant sucrose transporter polypeptides, and methods of utilizing the bacteria to produce products such as glycerol and glycerol-derived products are provided.

Variant sucrose transporter polypeptides that enable faster sucrose utilization in bacteria

Recombinant() bacteria that have enhanced acetyl-CoA synthetase activity and the ability to produce glycerol and glycerol-derived products, such as 3-hydroxypropionic acid, methylglyoxal, 1,2-propanediol, and 1,3-propanediol, are described. The recombinantcomprise a promoter operably linked to a nucleotide sequence encoding a polypeptide having acetyl-CoA synthetase enzyme activity, wherein the promoter and nucleotide sequence are each independently either native or non-native.

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Disrupting the expression of endogenoushost cell genes gcvA and spr provides mutant host cells having increased heterologous peptide production. The addition of a genetic modification to the coding region of gene yejM further enhances peptide production and facilitates easier downstream processing. Recombinanthost cells are provided as well as methods of using such host cells for heterologous peptide production.

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Wallingford, PA, United States of America

Qi Chen