Engineered Escherichia coli for simultaneous utilization of galactose and glucose

Lim, Hyun Gyu; Seo, Sang Woo; Jung, Gyoo Yeol

doi:10.1016/j.biortech.2012.10.124

Cited by 34 publications

(36 citation statements)

References 20 publications

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“…Then, all genetic parts including promoters, 5′-UTRs, coding sequences, and terminators were assembled as a pseudo-operon in a plasmid (pACYC-Lys). Subsequently, this artificial operon was integrated into the chromosome as an additional copy by replacing galR to increase the galactose utilization rate [15]. For efficient recombination, lysA , which is next to galR in the genome, was also deleted.…”

Section: Resultsmentioning

confidence: 99%

“…2 a Specific growth rate and b maximum specific galactose uptake rate of DHK 1–3 strains. One OD 600 unit corresponds to 0.27 g dry cell weight (DCW)/L [15]. c – e Time-course fermentation profiles of DHK1-3 strains.…”

Section: Resultsmentioning

confidence: 99%

“…Thus, it is expected that galactose can be a suitable feedstock to produce various platform chemicals in large quantities. However, the major drawback of industrial microorganisms such as E. coli is the slower utilization rate of galactose than that of glucose [15]. This low assimilation rate of galactose results in reduced rates of both growth and product formation even with well-performed glucose-dependent production pathways [16].…”

Section: Introductionmentioning

confidence: 99%

“…To overcome the limitation on galactose utilization, several combinatorial approaches have been demonstrated such as expression of several combinations of metabolic genes on C. glutamicum [17] and construction of fragmented chromosomal library perturbations for inverse metabolic engineering as well as a random mutagenesis approach on Saccharomyces cerevisiae [18, 19]. In a recent study, E. coli was re-designed by reconstruction of its utilization pathway with synthetic genetic parts including predictable promoters, 5′-untranslated regions (5′-UTRs), and terminators to achieve maximum expression [15]. The engineered strain showed a significantly enhanced growth rate (44.8%) and sugar utilization rate (53.1%), similar to glucose fermentation [15].…”

Section: Introductionmentioning

confidence: 99%

“…In a recent study, E. coli was re-designed by reconstruction of its utilization pathway with synthetic genetic parts including predictable promoters, 5′-untranslated regions (5′-UTRs), and terminators to achieve maximum expression [15]. The engineered strain showed a significantly enhanced growth rate (44.8%) and sugar utilization rate (53.1%), similar to glucose fermentation [15]. Furthermore, this engineered pathway was shown to be efficient when combined with the n-butanol production pathway [20].…”

Section: Introductionmentioning

confidence: 99%

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Synthetic redesign of Escherichia coli for cadaverine production from galactose

Kwak

Lim

Yang

et al. 2017

Biotechnol Biofuels

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BackgroundWith increasing concerns over the environment, biological production of cadaverine has been suggested as an alternative route to replace polyamides generated from the petroleum-based process. For an ideal bioprocess, cadaverine should be produced with high yield and productivity from various sugars abundant in biomass. However, most microorganisms are not able to efficiently metabolize other biomass-derived sugars as fast as glucose. This results in reduced growth rate and low carbon flux toward the production of desired bio-chemicals. Thus, redesign of microorganisms is necessary for utilizing those carbon sources with enhanced carbon flux and product formation.ResultsIn this study, we engineered Escherichia coli to produce cadaverine with rapid assimilation of galactose, a promising future feedstock. To achieve this, genes related to the metabolic pathway were maximally expressed to amplify the flux toward cadaverine production via synthetic expression cassettes consisting of predictive and quantitative genetic parts (promoters, 5′-untranslated regions, and terminators). Furthermore, the feedback inhibition of metabolic enzymes and degradation/re-uptake pathways was inactivated to robustly produce cadaverine. Finally, the resultant strain, DHK4, produced 8.80 g/L cadaverine with high yield (0.170 g/g) and productivity (0.293 g/L/h) during fed-batch fermentation, which was similar to or better than the previous glucose fermentation.ConclusionsTaken together, synthetic redesign of a microorganism with predictive and quantitative genetic parts is a prerequisite for converting sugars from abundant biomass into desired platform chemicals. This is the first report to produce cadaverine from galactose. Moreover, the yield (0.170 g/g) was the highest among engineered E. coli systems.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-017-0707-2) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Synthetic redesign of Escherichia coli for cadaverine production from galactose

Kwak

Lim

Yang

et al. 2017

Biotechnol Biofuels

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Co‐utilization of hexoses by a microconsortium of sugar‐specific E. coli strains

Chappell

Nair

2017

Biotech & Bioengineering

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Escherichia coli is an important commercial species used for production of biofuels, biopolymers, organic acids, sugar alcohols, and natural compounds. Processed biomass and agroindustrial byproducts serve as low-cost nutrient sources and contain a variety of hexoses available for bioconversion. However, metabolism of hexose mixtures by E. coli is inefficient due to carbon catabolite repression (CCR), where the transport and catabolic activity of one or more carbon sources is repressed and/or inhibited by the transport and catabolism of another carbon source. In this work, we developed a microconsortium of different E. coli strains, each engineered to preferentially catabolize a different hexose-glucose, galactose, or mannose. We modified the specificity and preference of carbon source using a combination of rational strain design and adaptive evolution. The modifications ultimately resulted in strains that preferentially catabolized their specified sugar. Finally, comparative analysis in galactose- and mannose-rich sugar mixtures revealed that the consortium grew faster and to higher cell densities compared to the wild-type strain. Biotechnol. Bioeng. 2017;114: 2309-2318. © 2017 Wiley Periodicals, Inc.

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Microbial Production of Amines and Amino Acids by Fermentation

Wendisch

Kerbs

2022

Microbial Production of High-Value Products

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Engineered Escherichia coli for simultaneous utilization of galactose and glucose

Cited by 34 publications

References 20 publications

Synthetic redesign of Escherichia coli for cadaverine production from galactose

Synthetic redesign of Escherichia coli for cadaverine production from galactose

Co‐utilization of hexoses by a microconsortium of sugar‐specific E. coli strains

Microbial Production of Amines and Amino Acids by Fermentation

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