E.i. Dupont De Nemours and Company

Nutrition & Biosciences USA 4, Inc.

Kristin Ruebling-Jass

Qiong Cheng

Jian Ping Lai

Qi Chen

Zheng You

Robert DiCosimo

Mark Scott Payne

Steven Cary Rothman

Arthur Ouwehand

Kevin Michael Croker

Jianzhong Zhang

Hong Peng Wang

Pierre E Rouviere

Andrew C Eliot

Stephen R Fahnestock

Jean-Francois Tomb

Tina K Van Dyk

Tanja Maria Gruber

Thomas Eduard Friedmann

Annapurna Sachan

Ritu Bhalla

Neeraj Pandey

An enzymatically produced soluble α-glucan fiber composition is provided suitable for use as a digestion resistant fiber in food and feed applications. The soluble α-glucan fiber composition can be blended with one or more additional food ingredients to produce fiber-containing compositions. Methods for the production and use of compositions comprising the soluble α-glucan fiber are also provided.

Enzymatic synthesis of soluble glucan fiber

An enzymatically produced soluble &#x3b1;-glucan fiber composition is provided suitable for use as a digestion resistant fiber in food and feed applications. The soluble &#x3b1;-glucan fiber composition can be blended with one or more additional food ingredients to produce fiber-containing compositions. Methods for the production and use of compositions comprising the soluble &#x3b1;-glucan fiber are also provided.

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 capable of metabolizing sucrose are described. The recombinant bacteria comprise in their genome or on at least one recombinant construct, a novel nucleotide sequence encoding a polypeptide having sucrose transporter activity and a nucleotide sequence encoding a polypeptide having sucrose hydrolase activity. These nucleotide sequences are each operably linked to the same or a different promoter. Recombinant bacteria capable of metabolizing sucrose to produce glycerol and/or glycerol-derived products such as 1,3-propanediol and 3-hydroxypropionic acid are also described.

Recombinant bacteria comprising novel sucrose transporters

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.

Host cell modifications that improve peptide production and downstream processing

A process of in vivo labeling and identifying recombinantly produced peptides or proteins within an unpermeabilized prokaryotic host cell. Recombinant prokaryotic cells expressing a fusion peptide comprising at least one tetracysteine tag were labeled in vivo using a biarsenical labeling reagent. A fluorescent activated cell sorter was used to identify and select subpopulations of fluorescent cells wherein the amount of fusion peptide in the cell was proportional to the amount of fluorescence detected.

Use of tetracysteine tags in fluorescence-activated cell sorting analysis of prokaryotic cells producing peptides or proteins

Disrupting the expression of endogenoushost cell genes gcvA and spr provides mutant host cells having increased heterologous peptide production relative to controlhost cells. Recombinanthost cells are provided as well as methods of using such host cells for heterologous peptide production.

Growing community of inventors

Wilmington, DE, United States of America

Kristin Ruebling-Jass