Maximum Volumetric Ethanol Productivity Research Articles

Fermentation of lignocellulosic hydrolyzate using a submerged membrane bioreactor at high dilution rates.

A submerged membrane bioreactor (sMBR) was developed to ferment toxic lignocellulosic hydrolyzate to ethanol. The sMBR achieved high cell density of Saccharomyces cerevisiae during continuous cultivation of the hydrolyzate by completely retaining all yeast cells inside the sMBR. The performance of the sMBR was evaluated based on the ethanol yield and productivity at the dilution rates 0.2, 0.4, 0.6, and 0.8h(-1) with the increase of dilution rate. Results show that the yeast in the sMBR was able to ferment the wood hydrolyzate even at high dilution rates, attaining a maximum volumetric ethanol productivity of 7.94 ± 0.10 g L(-1)h(-1) at a dilution rate of 0.8h(-1). Ethanol yields were stable at 0.44 ± 0.02 g g(-1) during all the tested dilution rates, and the ethanol productivity increased from 2.16 ± 0.15 to 7.94 ± 0.10 g L(-1)h(-1). The developed sMBR systems running at high yeast density demonstrate a potential for a rapid and productive ethanol production from wood hydrolyzate.

Ethanol Production from Starch by Co-Immobilized Amyloglucosidase—Zymomonas Mobilis Cells in a Continuously-Stirred Bioreactor

ABSTRACTContinuous ethanol production from starch was studied in a continuously-stirred bioreactor using co-immobilized amyloglucosidase (AMG) and Zymomonas mobilis cells. In this process, saccharification of starch and fermentation to ethanol were carried out in a single reactor.The experiments were carried out at three sets of conditions for which temperature and centrifuged cell weight were 35°C and 20.3 g; 30°C and 19.0 g; 35°C and 29.2 g, respectively. The maximum volumetric ethanol productivity (7.6 g·L−1h−1) was achieved with 0.45 g·g−1 yield at the 0.20 h−1 dilution rate at 35°C, with the initial centrifuged cell concentration of 29.2 g. The maximum ethanol concentration (43.5 g L−1) was obtained at the 0.15 h−1 dilution rate with a 96 % conversion of substrate.When experimental conditions in the reactor were compared to each other, the initial concentration of immobilized cells was found to have a significant positive influence on the kinetic parameters of ethanol fermentation.

Ethanol production from concentrated food waste hydrolysates with yeast cells immobilized on corn stalk

The aim of the present study was to examine ethanol production from concentrated food waste hydrolysates using whole cells of S. cerevisiae immobilized on corn stalks. In order to improve cell immobilization efficiency, biological modification of the carrier was carried out by cellulase hydrolysis. The results show that proper modification of the carrier with cellulase hydrolysis was suitable for cell immobilization. The mechanism proposed, cellulase hydrolysis, not only increased the immobilized cell concentration, but also disrupted the sleek surface to become rough and porous, which enhanced ethanol production. In batch fermentation with an initial reducing sugar concentration of 202.64 ± 1.86 g/l, an optimal ethanol concentration of 87.91 ± 1.98 g/l was obtained using a modified corn stalk-immobilized cell system. The ethanol concentration produced by the immobilized cells was 6.9% higher than that produced by the free cells. Ethanol production in the 14th cycle repeated batch fermentation demonstrated the enhanced stability of the immobilized yeast cells. Under continuous fermentation in an immobilized cell reactor, the maximum ethanol concentration of 84.85 g/l, and the highest ethanol yield of 0.43 g/g (of reducing sugar) were achieved at hydraulic retention time (HRT) of 3.10 h, whereas the maximum volumetric ethanol productivity of 43.54 g/l/h was observed at a HRT of 1.55 h.

Fuel ethanol production from concentrated food waste hydrolysates in immobilized cell reactors by Saccharomyces cerevisiae H058

BACKGROUND: Continuous ethanol fermentation of concentrated food waste hydrolysates has been studied. The process was carried out in an immobilized cell reactor with beads of calcium-alginate containing immobilized Saccharomyces cerevisiae H058 at temperature 30 °C and pH 5.0. RESULTS: The total residual sugar decreased with increase of hydraulic retention time (HRT) under various reducing sugar concentrations. Ethanol production by immobilized cells increased with increase in HRT, regardless of the substrate concentrations employed. The highest ethanol concentration of 89.28 g L−1 was achieved at an HRT of 5.87 h and reducing sugar concentration of 200 g L−1. At an HRT of 1.47 h, the maximum volumetric ethanol productivity of 49.88 g L−1 h−1 and the highest ethanol yield of 0.48 g g−1 were achieved at reducing sugar concentration of 160 and 200 g L−1, respectively. The difference between the fresh and the 30-day Ca–alginate immobilized cell was also shown by scanning electronic micrographs of beads taken from their outer and inner surfaces. CONCLUSIONS: Continuous ethanol production from concentrated food waste hydrolysates using immobilized yeast cells is promising in view of the high ethanol productivity obtained at relatively high conversion and excellent reactor stability. Copyright © 2011 Society of Chemical Industry

Performance testing of Zymomonas mobilis metabolically engineered for cofermentation of glucose, xylose, and arabinose.

IOGEN Corporation of Ottawa, Canada, has recently built a 40t/d biomass-to-ethanol demonstration plant adjacent to its enzyme production facility. It has partnered with the University of Toronto to test the C6/C5 cofermenta-tion performance characteristics of the National Renewable Energy Labora-tory's metabolically engineered Zymomonas mobilis using various biomass hydrolysates. IOGEN's feedstocks are primarily agricultural wastes such as corn stover and wheat straw. Integrated recombinant Z. mobilis strain AX101 grows on D-xylose and/or L-arabinose as the sole carbon/energy sources and ferments these pentose sugars to ethanol in high yield. Strain AX101 lacks the tetracycline resistance gene that was a common feature of other recombinant Zm constructs. Genomic integration provides reliable cofermentation performance in the absence of antibiotics, another characteristic making strain AX101 attractive for industrial cellulosic ethanol production. In this work, IOGEN's biomass hydrolysate was simulated by a pure sugar medium containing 6% (w/v) glucose, 3% xylose, and 0.35% arabinose. At a level of 3 g/L (dry solids), corn steep liquor with inorganic nitrogen (0.8 g/L of ammonium chloride or 1.2 g/L of diammonium phosphate) was a cost-effective nutritional supplement. In the absence of acetic acid, the maximum volumetric ethanol productivity of a continuous fermentation at pH 5.0 was 3.54 g/L x h. During prolonged continuous fermentation, the efficiency of sugar-to-ethanol conversion (based on total sugar load) was maintained at >85%. At a level of 0.25% (w/v) acetic acid, the productivity decreased to 1.17 g/L x h at pH 5.5. Unlike integrated, xylose-utilizing rec Zm strain C25, strain AX101 produces less lactic acid as byproduct, owing to the fact that the Escherichia coli arabinose genes are inserted into a region of the host chromosome tentatively assigned to the gene for D-lactic acid dehydrogenase. In pH-controlled batch fermentations with sugar mixtures, the order of sugar exhaustion from the medium was glucose followed by xylose and arabinose. Both the total sugar load and the sugar ratio were shown to be important determinants for efficient cofermentation. Ethanol at a level of 3% (w/v) was implicated as both inhibitory to pentose fermentation and as a potentiator of acetic acid inhibition of pentose fermentation at pH 5.5. The effect of ethanol may have been underestimated in other assessments of acetic acid sensitivity. This work underscores the importance of employing similar assay conditions in making comparative assessments of biocatalyst fermentation performance.

Performance of a cross-linked immobilized cell reactor with Zymomonas mobilis using synthetic and complex media for ethanol production

The performance of ethanolic bacterium, Zymomonas mobilis, ATCC 31821, cross-linked to glutaraldehyde-treated, gelatin-coated spherical glass beads in an immobilized cell reactor (ICR) was evaluated for feed glucose concentrations of 150 and 200 g l −1. A synthetic medium, containing inorganic nitrogen and salts, and a complex medium, containing yeast extract, were both used. Two acid-base buffers, succinate and acetate, were added to the medium to regulate the pH of the broth in the ICR. The use of acetate in the broth increased the maximum volumetric ethanol productivity to 19 g l −1 h −1 compared to 16 gl −1 h −1 without acetate. Adding acetate to the medium increased the reactor operating life by decreasing cell growth rate. The reactors were operated from 34–57 days. The ICR fermentations conducted with the synthetic medium gave a comparable maximum volumetric ethanol productivity of 16 gl −1 h −1. Axial temperature distributions of 4–6°C were measured during the fermentation.

Ethanol production from glucose and xylose by separated and co-culture processes using high cell density systems

Media containing xylose and/or glucose were tested utilizing Zymomonas mobilis or Saccharomyces diastaticus and Pichia stipitis. The best fermentation results were obtained in separated glucose (180 g/litre) and xylose (80 g/litre) fermentations utilizing Zymomonas mobilis and Pichia stipitis strains, respectively. In these conditions, the maximum ethanol concentrations achieved were 86·2 g/litre and 29 g/litre, respectively. The complete conversion of a glucose and xylose mixture (50 g/litre) was obtained using a respiratory deficient mutant of Saccharomyces diastaticus co-cultivated with Pichia stipitis in continuous culture. Using the co-culture process, the maximum ethanol concentration was 21·5 g/litre (Yp/s=0·45 g/g) and the maximum volumetric ethanol productivity was 4·3 g/(litre × h).

Xylose fermentation by Candida shehatae and Pichia stipitis: effects of pH, temperature and substrate concentration

The effects of temperature, pH and xylose concentration on the fermentation parameters of Candida shehatae and Pichia stipitis were evaluated. The optimum pH was in the region of pH 4–5.5, with an optimum fermentation temperature of 30°C. Maximum fermentation rates were reached at 50 g l −1 xylose. A maximum volumetric ethanol productivity of about 0.9 g (l h) −1 was obtained with both yeast strains. The ethanol yield of C. shehatae decreased considerably when cultivated above 30°C or when the xylose concentration was increased. Xylitol accumulated concomitantly. Xylitol production by P. stipitis was observed only during cultivation at 36°C. Whereas the ethanol yield of C. shehatae was usually about 75% of the theoretical maximum, it was 85–90% with P. stipitis.

Aerobic Fermentation of D-Xylose to Ethanol by Clavispora sp

Eleven strains of an undescribed species of Clavispora fermented D-xylose directly to ethanol under aerobic conditions. Strain UWO(PS)83-877-1 was grown in a medium containing 2% D-xylose and 0.5% yeast extract, and the following results were obtained: ethanol yield coefficient (ethanol/D-xylose), 0.29 g g (57.4% of theoretical); cell yield coefficient (dry biomass/D-xylose), 0.25 g g; maximum ethanol concentration, 5.9 g liter; maximum volumetric ethanol productivity, 0.11 g liter h. With initial D-xylose concentrations of 40, 60, and 80 g liter, maximum ethanol concentrations of 8.8, 10.9, and 9.8 g liter were obtained, respectively (57.2, 57.1, and 48.3% of theoretical). Ethanol was found to inhibit the fermentation of D-xylose (K(p) = 0.58 g liter) more than the fermentation of glucose (K(p) = 6.5 g liter). The performance of this yeast compared favorably with that reported for some other D-xylose-fermenting yeasts.

Continuous ethanol production from Jerusalem artichoke tubers. II. Use of immobilized cells of Kluyveromyces marxianus

Kluyveromyces marxianus UCD (FST) 55-82 cells were immobilized in Na alginate beads and used in a packed-bed bioreactor system for the continuous production of ethanol from the extract of Jerusalem artichoke tubers. Volumetric ethanol productivities of 104 and 80 g ethanol/ L/h were obtained at 80 and 92% sugar utilization, respectively. The maximum volumetric ethanol productivity of the immobilized cell bioreactor system was found to be 15 times higher than that of an ordinary-stirred-tank (CST) bioreactor using cells of K. marxianus. The immobilized cell bioreactor system was operated continuously at a constant dilution rate of 0.66 h(-1) for 12 days resulting in only an 8% loss of the original immobilized cell activity, which corresponds to an estimated half-life of ca. 72 days. The maximum specific ethanol productivity and maximum specific sugar uptake rate of the immobilized cells were found to be 0.55 g ethanol/g/biomass/h and 1.21 g sugars/g biomass/h, respectively.