High yield production of d-xylonic acid from d-xylose using engineered Escherichia coli

Liu, Huaiwei; Nisola, Grace M.; Ramos, Kristine Rose M.; Chung, Wook-Jin

doi:10.1016/j.biortech.2011.08.065

Cited by 111 publications

(78 citation statements)

References 18 publications

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“…Previous studies have shown that deletion of these three genes disables the strain's capability to grow on D-xylose or D-xylonic acid but does not interfere with cellular growth on LB or glucose (Liu et al, 2012;Liu et al, 2013). These results indicated that the strain's central metabolism was not influenced by the ϭϭ blockage of D-xylose metabolism, suggesting that the BD pathway can be introduced to the chassis.…”

Section: Construction and Optimization Of The Enzymatic Reactor Modulementioning

confidence: 77%

“…The D-xylose dehydrogenase from Caulobacter crescentus (Xdh) is highly efficient for D-xylose oxidation and has been adopted for producing D-xylonic acid in both engineered E. coli and yeast strains previously (Liu et al, 2012;Toivari et al, 2012). E. coli K-12 family strains have two native D-xylonic acid dehydratases, YjhG and YagF, which can convert D-xylonic acid into 2-dehydro-3-deoxy-Dxylonate.…”

Section: Designing a De Novo Bd Pathway And An Associated Cellular Chmentioning

confidence: 99%

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Autonomous production of 1,4-butanediol via a de novo biosynthesis pathway in engineered Escherichia coli

Liu

2015

Metabolic Engineering

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105

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Section: Construction and Optimization Of The Enzymatic Reactor Modulementioning

confidence: 77%

Section: Designing a De Novo Bd Pathway And An Associated Cellular Chmentioning

confidence: 99%

Autonomous production of 1,4-butanediol via a de novo biosynthesis pathway in engineered Escherichia coli

Liu

2015

Metabolic Engineering

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105

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“…Although wild type bacteria are efficient in the xylonic acid production, they still have high nutritional requirements, and low cell biomass production yields, which makes their utilization in industrial processes difficult. Consequently, for the last three years, genetic engineering strategies were used to build recombinant xylonic acid producing strains of E. coli, S. cerevisiae, Kluyveromyces lactis and Pichia kudriavzevii [50][51][52][53][54][55]. These microorganisms were chosen as possible hosts for presenting high growth rates, simple nutritional requirements and specially yeasts, for presenting good tolerance to inhibitors found in lignocellulosic hydrolysates, as commented above [45].…”

Section: New Productsmentioning

confidence: 99%

“…These microorganisms were chosen as possible hosts for presenting high growth rates, simple nutritional requirements and specially yeasts, for presenting good tolerance to inhibitors found in lignocellulosic hydrolysates, as commented above [45]. Indeed, the identification of genes from different microorganisms, that code for the enzymes involved in the conversion of xylose to xylonic acid allowed construction of strains able to produce xylonic acid with yields above 90% of theoretical maximum and at high concentrations [50][51][52][53][54][55].…”

Section: New Productsmentioning

confidence: 99%

Genetic improvement of microorganisms for applications in biorefineries

Paes

Almeida

2014

Chem. Biol. Technol. Agric.

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The development of biorefineries directed to the production of fuels, chemicals and energy is important to reduce economic dependence and environmental impacts of a petroleum-based economy. Microorganisms are essential in several industrial bioprocesses nowadays, and it is expected that new microbial bioprocesses will play a key role in biorefineries. However, the bioconversion process requires a robust and highly productive microorganism. In this scenario, several strategies to genetically improve microorganisms to overcome the bioprocesses challenges have been considered. In this work, we review microorganisms importance in the biorefineries concept, highlight the desirable traits they must hold in order to be employed, and discuss the main strategies to improve such traits. The focuses of this work are on four main targets in the improvement of microorganisms: driving carbon flux towards the desired pathway, increasing tolerance to toxic compounds, increasing substrate uptake range and new products generation.

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“…Recently, engineered strains of Escherichia coli (8), Saccharomyces cerevisiae (9), and Pichia kudriavzevii (10) were described, which produce xylonic acid efficiently at a laboratory scale using a xylose dehydrogenase from Caulobacter crescentus (39.2 g/liter xylonic acid from 40 g/liter xylose [E. coli]; 43 g/liter xylonic acid from 49 g/liter xylose [S. cerevisiae]; and 146 g/liter xylonic acid from 153 g/liter xylose [P. kudriavzevii]). Fungal species are preferred industrial production organisms because of their low nutrition requirements and tolerance to growth inhibitors in lignocellulosic hydrolysates.…”

mentioning

confidence: 99%

Single-Cell Measurements of Enzyme Levels as a Predictive Tool for Cellular Fates during Organic Acid Production

Zdraljevic¹,

Wagner²,

Cheng³

et al. 2013

Appl Environ Microbiol

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bOrganic acids derived from engineered microbes can replace fossil-derived chemicals in many applications. Fungal hosts are preferred for organic acid production because they tolerate lignocellulosic hydrolysates and low pH, allowing economic production and recovery of the free acid. However, cell death caused by cytosolic acidification constrains productivity. Cytosolic acidification affects cells asynchronously, suggesting that there is an underlying cell-to-cell heterogeneity in acid productivity and/or in resistance to toxicity. We used fluorescence microscopy to investigate the relationship between enzyme concentration, cytosolic pH, and viability at the single-cell level in Saccharomyces cerevisiae engineered to synthesize xylonic acid. We found that cultures producing xylonic acid accumulate cells with cytosolic pH below 5 (referred to here as "acidified"). Using live-cell time courses, we found that the probability of acidification was related to the initial levels of xylose dehydrogenase and sharply increased from 0.2 to 0.8 with just a 60% increase in enzyme abundance (Hill coefficient, >6). This "switch-like" relationship likely results from an enzyme level threshold above which the produced acid overwhelms the cell's pH buffering capacity. Consistent with this hypothesis, we showed that expression of xylose dehydrogenase from a chromosomal locus yields ϳ20 times fewer acidified cells and ϳ2-fold more xylonic acid relative to expression of the enzyme from a plasmid with variable copy number. These results suggest that strategies that further reduce cell-to-cell heterogeneity in enzyme levels could result in additional gains in xylonic acid productivity. Our results demonstrate a generalizable approach that takes advantage of the cell-to-cell variation of a clonal population to uncover causal relationships in the toxicity of engineered pathways.

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High yield production of d-xylonic acid from d-xylose using engineered Escherichia coli

Cited by 111 publications

References 18 publications

Autonomous production of 1,4-butanediol via a de novo biosynthesis pathway in engineered Escherichia coli

Autonomous production of 1,4-butanediol via a de novo biosynthesis pathway in engineered Escherichia coli

Genetic improvement of microorganisms for applications in biorefineries

Single-Cell Measurements of Enzyme Levels as a Predictive Tool for Cellular Fates during Organic Acid Production

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