High succinate producing bacteria

A technology of bacteria and cells, applied in the direction of bacteria, fermentation, etc., can solve problems such as limitations

Inactive Publication Date: 2007-08-22
RICE UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods are limited by the lack of succinate production during the aerobic phase and the stringent conditions of the anaerobic growth phase for succinate production

Method used

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Embodiment approach

[0016] The carboxylic acids may be salts, acids, bases or derivatives, depending on structure, pH and ions present. For example, the terms "succinate" and "succinic acid" are used interchangeably herein. Succinic acid is also called succinic acid (C 4 h 6 o 4 ). The chemicals used here included formic acid, glyoxylic acid, lactic acid, malic acid, oxaloacetic acid (OAA), phosphoenolpyruvate (PEP) and pyruvate. Bacterial metabolic pathways including the Krebs cycle (also known as the citric acid, tricarboxylic acid, or TCA cycle) can be found in Lehninger's "Principles of Biochemistry" and other biochemistry textbooks.

[0017] The terms "operably associated" or "operably linked" are used to refer to functionally paired nucleotide sequences.

[0018] "Reduced activity" or "inactivation" as defined herein means at least a 75% reduction in the activity of the protein compared to an appropriate control strain. Preferably, at least 80, 85, 90, 95% reduction in activity is ach...

Embodiment 1

[0031] Example 1: Inhibition of Lactic Acid, Acetic Acid and Ethanol

[0032] Hybrid bacterial strains that produce carboxylic acids under aerobic and anaerobic conditions are able to overcome the limitations of low biomass generation on anaerobic production processes. Biomass can be produced under aerobic conditions at the beginning of the fermentation process. During this period, a large number of carboxylic acids are generated through the aerobic synthetic pathway, saving time and cost. Once high biomass is achieved, the environment can be switched or allowed to switch to anaerobic conditions for additional conversion of carbon sources to carboxylic acids in high yield. Using a redesigned anaerobic succinate fermentation pathway, carboxylic acid production is expected to increase to well above 2 or 3 moles of product per mole of glucose.

[0033] First, to increase flux into the TCA cycle for carboxylic acid production, two acetate pathways in aerobic metabolism are ina...

Embodiment 2

[0037] Example 2: Increasing glyoxylate bypass flux

[0038] As previously shown, the presence of native ACEA and ACEB is sufficient to drive carboxylic acid production without additional expression. However, native expression levels are susceptible to feedback inhibition and are sensitive to the aerobic or anaerobic conditions of the environment. Constitutive activation of the glyoxylate shunt is important to maintain high levels of aerobic metabolism for carboxylic acid synthesis. This activation is made possible by inactivating the aceBAK operator repressor (ICLR). As shown in Figure 1, activation of the glyoxylate bypass provides a mixed fermentation environment that achieves high levels of carboxylic acid production.

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Abstract

The invention relates to a hybrid succinate production system that has a high capacity to produce succinate under aerobic and anaerobic conditions. The metabolic engineering of a hybrid bacterial succinate production system that can function under both aerobic and anaerobic conditions makes the production process more efficient, and the process control and optimization less difficult.

Description

field of invention [0001] The present invention relates to a hybrid succinate producing system designed in E. coli and designed to produce high levels of succinate under both aerobic and anaerobic conditions. Background of the invention [0002] The expensive specialty chemical succinic acid and its derivatives have a wide range of industrial applications. Succinic acid is used as a raw material in food, pharmaceuticals, plastics, cosmetics and textiles, and in electroplating and waste-gas scrubbing (61). Succinic acid can be used as a raw material for some plastic precursors, such as 1,4-butanediol (BDO), tetrahydrofuran and gamma-butyrolactone. Also, succinic acid and BDO can be used as monomers for polyester. If the cost of succinic acid could be reduced, it would become very useful as an intermediate feedstock for the production of other bulk chemicals (47). In addition to succinic acid, other 4-carbon dicarboxylic acids such as malic acid and fumaric acid also have r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/46C12N1/20
CPCC12P7/46C12P7/40C12N1/20
Inventor K-Y·单乔治·N·贝内特亨利·林艾伦·桑切斯
Owner RICE UNIV
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