J.M. Navarro

Effects of lignocellulose degradation products on ethanol fermentations of glucose and xylose by Saccharomyces cerevisiae, Zymomonas mobilis, Pichia stipitis, and Candida shehatae

The inhibitory effects of six lignocellulose degradation products on glucose fermentation by Saccharomyces cerevisiae and Zymomonas mobilis on xylose fermentation by Pichia stipitis and Candida shehatae were studied in batch cultures. Toxic compounds were added in varying concentrations and subsequent inhibitions on growth and ethanol production were quantified. Vanillin was shown to be a strong inhibitor of both growth and ethanol production by xylose fermenting yeasts and S. cerevisiae when it was added to the culture media at a concentration of 1 g l−1. Fermentative activities of Z. mobilis were greatly sensitive to the presence of hydroxybenzaldehyde (0.5 g l−1). Analysis of culture media extracts showed that some of the inhibitors, particularly vanillin and furaldehyde, could be assimilated by the tested microbial strains which resulted in the partial recovery in both growth and ethanol production processes on prolonged incubation.

Xylitol production from D-xylose byCandida guillermondii: Fermentation behaviour

SummaryThe ability ofCandida guillermondii to produce xylitol from xylose and to ferment individual non xylose hemicellulosic derived sugars was investigated in microaerobic conditions. Xylose was converted into xylitol with a yield of 0,63 g/g and ethanol was produced in negligible amounts. The strain did not convert glucose, mannose and galactose into their corresponding polyols but only into ethanol and cell mass. By contrast, fermentation of arabinose lead to the formation of arabitol. On D-xylose medium,Candida guillermondii exhibited high yield and rate of xylitol production when the initial sugar concentration exceeded 110 g/l. A final xylitol concentration of 221 g/l was obtained from 300 g/l D-xylose with a yield of 82,6% of theoretical and an average specific rate of 0,19 g/g.h.

On-line automated monitoring and control systems for CO2 and O2 in aerobic and anaerobic solid-state fermentations

Abstract Two systems for monitoring and control of gases from solid-state cultures have been developed. The first involves on-line automated monitoring of CO 2 and O 2 concentration in exhaust gases from eight fermenters or eight gas sampling ports in a large fermenter. It proved to be efficient in obtaining information on the physiological state and respiration rate of the culture in a real-time process. Furthermore, the specific growth rate (μ) can be estimated reliably by gas measurements in aerobic cultures. The second system is for automated control of exit gases from aerobic solid-state fermentations. It permitted elimination of biomass and temperature gradients in a large fermenter due to the maintenance of culture under non-limiting conditions on oxygen. These two systems have applicability in aerobic and anaerobic solid-state processes and were found to be reliable in a number of fermentation experiments as well as optimization of solid-state fermentation. To our knowledge no earlier report of such versatile and reliable on-line automated monitoring/control systems has appeared.

Xylitol production from xylose by two yeast strains: Sugar tolerance

The kinetics and enzymology ofd-xylose utilization are studied in micro-, semi-, and aerobic batch cultures during growth ofCandida guilliermondii andCandida parapsilosis in the presence of several initial xylose concentrations. The abilities of xylitol accumulation by these two yeast strains are high and similar, although observed under various growth conditions. WithCandida parapsilosis, optimal xylitol production yield (0.74 g/g) was obtained in microaerobiosis with 100 g/L of xylose, whereas optimal conditions to produce xylitol byCandida guilliermondii (0.69 g/g) arose from aerobiosis with 300 g/L of sugar. The different behavior of these yeasts is most probably explained by differences in the nature of the initial step of xylose metabolism: a NADPH-linked xylose reductase activity is measured with a weaker NADH-linked activity. These activities seem to be dependent on the degree of aerobiosis and on the initial xylose concentration and correlate with xylitol accumulation.

Maintenance of Heat and Water Balances as a Scale-up Criterion for the Production of Ethanol by Schwanniomyces castellii in a Solid State Fermentation System

The scale-up of column fermenters [for ethanol fermentation from sugar cane bagasse by Schwanniomyces castellii CBS 2863] by 6 to 410 gravimetric scale factors from 10 g moist substrate size was achieved efficiently by maintaining heat and water balances in the media. Data on patterns of ethanol production, biomass formation, the concn. of different carbohydrates, DM and pH values against time were of equal magnitude in 10- and 60-g size column fermenters. Reactors of 60-, 370- and 4100-g sizes also compared well in respect of O2 consumption, CO2 evolution and the specific growth rates in aerobic and anaerobic fermentation phases. Overall productivities of ethanol were similar in all the 4 column fermenters. The ability to obtain the same results in all the fermenter sizes, in spite of increases in diam. and the height of the columns, indicates the high potential of this simple scale-up criterion which has not been used earlier for scale-up of any fermentation process.

The effect of aeration on D-xylose fermentation by Pachysolen tannophilus, Pichia stipitis, Kluyveromyces marxianus and Candida shehatae.

SummaryThe fermentation of D-xylose byPachysolen tannophilus Y2460,Pichia stipitis Y7124,Kluyveromyces marxianus Y2415 andCandida shehatae Y12878 was investigated in aerobic, anaerobic and microaerophilic batch cultures. The aeration rate greatly influenced the fermentations; growth, rate of ethanol production and oxidation of ethanol are affected. Of the strains tested,Pichia stipitis appears superior; under anaerobic conditions it converts D-xylose (20 g/l) to ethanol with a yield of 0.40 g/l and it exhibits the highest ethanol specific productivity (3.5 g of ethanol per g dry cell per day) under microaerophilic conditions.

Production of alcohol from raw wheat flour by Amyloglucosidase and Saccharomyces cerevisiae

Ethanol production, by a simultaneous saccharification and fermentation process from raw wheat flour, has been performed by Saccharomyces cerevisiae and a low level of amyloglucosidase enzyme. The fermentation time was about 60 h after a 6 h pre-saccharification, with an amyloglucosidase (AMG) level of 270 AGU. kg(-1) starch, but only 31 h with a simultaneous saccharification fermentation process (SSF). When an AMG level of 540 AGU. kg(-1) starch was used, the time decreased to 21 h, giving an ethanol concentration of 67 g. l(-1). Sugar composition of the wort after the liquefaction may be responsible of the difference between these two process. Maltose, a fermentable sugar, was produced in high concentration during the liquefaction, allowing a shorter process period, counteracting the effect of the slow starch hydrolysis at 35 degrees C (SSF temperature).

Dielectric monitoring of growth and sporulation of Bacillus thuringiensis

On-line permittivity and optical density measurements have been used to monitor biomass concentration and sporulation status during growth of a spore-forming bacterium, Bacillus thuringiensis, in fed-batch culture. The correlation between permittivity, optical density and other observations showed three distinct phases of growth: growth itself, transition and sporulation. The permittivity variations during the transition and sporulation phases could be related to the sporulation development: the evolution pattern of the ratio of optical density to permittivity was representative of the culture state, and during the sporulation phase, a permittivity index could be build to measure the extend of spore liberation.

The reduction of xylose to xylitol by Candida guilliermondii and Candida parapsilosis: Incidence of oxygen and pH

SummaryThe ability of C. guilliermondii and C. parapsilosis to ferment xylose to xylitol was evaluated under different oxygen transfer rates in order to enhance the xylitol yield. In C. guilliermondii, a maximal xylitol yield of 0.66 g/g was obtained when oxygen transfer rate was 2.2 mmol/l.h. Optimal conditions to produce xylitol by C. parapsilosis (0.75 g/g) arose from cultures at pH 4.75 with 0.4 mmoles of oxygen/l.h. The response of the yeasts to anaerobic conditions has shown that oxygen was required for xylose metabolism.

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).