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Methanol Independent Expression by Pichia Pastoris Employing De-repression Technologies

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Published on: January 23, 2019

Homolactate fermentation by metabolically engineered Escherichia coli strains.

Y Zhu¹, M A Eiteman, K DeWitt

¹Center for Molecular BioEngineering, Department of Biological and Agricultural Engineering, University of Georgia, Athens, GA 30602, USA.

Applied and Environmental Microbiology

|November 24, 2006

Summary

This study engineered Escherichia coli for high-yield lactate production. Optimized strains achieved 138 g/L lactate with high efficiency, minimizing byproduct formation for industrial applications.

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Light-Controlled Fermentations for Microbial Chemical and Protein Production

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Published on: March 22, 2022

Area of Science:

Biotechnology
Metabolic Engineering
Microbial Fermentation

Background:

Lactate production is crucial for various industries.
Escherichia coli engineering offers a platform for efficient biochemical synthesis.
Optimizing fermentation pathways is key to maximizing product yield and purity.

Purpose of the Study:

To develop a high-yield, homofermentative lactate production process in engineered Escherichia coli.
To investigate and minimize succinate byproduct formation during fermentation.
To enhance lactate yield, purity, and productivity through genetic modification and process optimization.

Main Methods:

Genetic modification of Escherichia coli strains by knocking out key metabolic genes (aceEF, pfl, poxB, pps, frdABCD).
Two-phase fermentation: aerobic growth followed by anaerobic non-growth production.
Optimization of pH control using calcium hydroxide (Ca(OH)2).
(13)C nuclear magnetic resonance analysis to trace metabolic pathways and byproduct formation.

Main Results:

Engineered strain YYC202 achieved 90 g/L lactate with high yield and productivity.
Strain ALS974 (YYC202 frdABCD) reduced succinate accumulation by 70% and achieved 138 g/L lactate.
Optimized conditions with ALS974 yielded 0.99 g/g lactate with 97% carbon product purity and 6.3 g/L/h productivity.
(13)C NMR confirmed succinate derived from acetate, not the glyoxylate cycle.

Conclusions:

Homofermentative lactate production in engineered E. coli is feasible at high titers.
Minimizing succinate formation through gene knockout (frdABCD) and acetate control significantly improves lactate yield and purity.
This optimized process demonstrates a promising strategy for industrial-scale lactate manufacturing.