Ethanol production from xylose with a pyruvate-formate-lyase mutant of Klebsiella planticola carrying a pyruvate-decarboxylase gene from Zymomonas mobilis

Feldmann, Sigrun D.; Sprenger, Georg A.; Sahm, Hermann

doi:10.1007/bf00262454

Cited by 30 publications

(16 citation statements)

References 21 publications

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“…Our studies have confirmed previous reports (3,8,28,29) of poor ethanol production by enteric recombinants which contain only the Z. mobilis pdc gene. This poor performance is due in large part to the accumulation of acetaldehyde, indicating a requirement for additional ADH activity.…”

Section: Discussionsupporting

confidence: 91%

“…The accumulation of organic acids (formate and acetate) was proposed to contribute to increased ethanol toxicity. Formate and acetate production was blocked by the selec-tion of K. planticola pyruvate formate-lyase mutants (8). Although the elimination of this competing fermentation pathway improved ethanol production, the performance of these mutants remained below that observed previously with E. coli recombinants expressing both the Z. mobilis pdc and adhB genes (21).…”

mentioning

confidence: 83%

“…In principle, the transfer of Z. mobilis genes to other organisms could be used to produce recombinants with additional useful traits for ethanol production. Recombinants of Erwinia chrysanthemi (28) and Klebsiella planticola (8,29) have been constructed by using the Z. mobilis pdc gene alone. However, pyruvate metabolism was incompletely diverted to ethanol as a product of fermentation.…”

mentioning

confidence: 99%

“…The low expression of PDC from pZM15 was proposed as being insufficient to divert pyruvate metabolism. Higher ethanol yields were subsequently obtained with constructs which expressed higher levels of PDC (8). The accumulation of organic acids (formate and acetate) was proposed to contribute to increased ethanol toxicity.…”

mentioning

confidence: 99%

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Metabolic engineering of Klebsiella oxytoca M5A1 for ethanol production from xylose and glucose

Ohta

Beall

Mejia

et al. 1991

Appl Environ Microbiol

164

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The efficient diversion of pyruvate from normal fermentative pathways to ethanol production in Klebsiella oxytoca M5A1 requires the expression of Zymomonas mobilis genes encoding both pyruvate decarboxylase and alcohol dehydrogenase. Final ethanol concentrations obtained with the best recombinant, strain M5A1 (pLOI555), were in excess of 40 g/liter with an efficiency of 0.48 g of ethanol (xylose) and 0.50 g of ethanol (glucose) per g of sugar, as compared with a theoretical maximum of 0.51 g of ethanol per g of sugar. The maximal volumetric productivity per hour for both sugars was 2.0 g/liter. This volumetric productivity with xylose is almost twice that previously obtained with ethanologenic Escherichia coli. Succinate was also produced as a minor product during fermentation. * Corresponding author. t Florida Agricultural Experiment Station publication no. R01797.

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Section: Discussionsupporting

confidence: 91%

mentioning

confidence: 83%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Metabolic engineering of Klebsiella oxytoca M5A1 for ethanol production from xylose and glucose

Ohta

Beall

Mejia

et al. 1991

Appl Environ Microbiol

164

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“…However, it was noted that during TM89-pGOF111 fermentations at 45°C, no formate was produced but low level (± 185µM) fumarate accumulation was detected in culture supernatants. This may be due to a reduced metabolic flux through pfl where the active GoxPDC OPT may outcompete the pfl enzyme for pyruvate (Tolan et al, 1987;Feldmann et al, 1989;Orencio-Trejo et al, 2008).…”

Section: Expression Of Goxpdc Wt and Goxpdc Opt In E Coli And G Thementioning

confidence: 99%

Engineering pyruvate decarboxylase-mediated ethanol production in the thermophilic host Geobacillus thermoglucosidasius

Zyl

Taylor

Eley

et al. 2013

Appl Microbiol Biotechnol

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This study reports the expression, purification and kinetic characterization of a PDC from Gluconobacter oxydans. Kinetic analyses showed the enzyme to have high affinity for pyruvate (120µM at pH 5), high catalytic efficiency (4.75 x 10 5 M -1 s -1 at pH 5), a pH opt of approximately 4.5 and an in vitro temperature optimum at approximately 55°C (the highest yet reported for a b a c t e r i a l P D C ) . D u e t o g o o d in vitro thermostablity (approximately 40% enzyme activity retained after 30 minutes at 65°C) this PDC was considered to be a suitable candidate for heterologous expression in the thermophile Geobacillus thermoglucosidasius. Initial studies using a variety of methods failed to detect activity at any growth temperature. However, the application of codon harmonization (i.e., mimicry of the heterogeneous host's transcription and translational rhythm) yielded a protein that was fully functional in the thermophilic strain at 45°C (as determined by enzyme activity, Western blot, mRNA detection and ethanol productivity). Here we describe the successful expression of PDC in a true thermophile. Yields as high as 0.35 g/g ±0.04 ethanol per gram of glucose consumed were detected, highly competitive to those reported in ethanologenic thermophilic mutants. Although activities could not be detected at temperatures approaching the growth optimum for the strain, this study highlights that the possibility that previously unsuccessful expression of pdcs in Geobacillus spp. may be the result of ineffective transcription / translation coupling.

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Metabolic Design

Sahm¹

1993

Biotechnology

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Ethanol production from xylose with a pyruvate-formate-lyase mutant of Klebsiella planticola carrying a pyruvate-decarboxylase gene from Zymomonas mobilis

Cited by 30 publications

References 21 publications

Metabolic engineering of Klebsiella oxytoca M5A1 for ethanol production from xylose and glucose

Metabolic engineering of Klebsiella oxytoca M5A1 for ethanol production from xylose and glucose

Engineering pyruvate decarboxylase-mediated ethanol production in the thermophilic host Geobacillus thermoglucosidasius

Metabolic Design

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