Andreas Rudolf

Designing simultaneous saccharification and fermentation for improved xylose conversion by a recombinant strain of Saccharomyces cerevisiae.

Wheat straw is an abundant agricultural residue which can be used as a raw material for bioethanol production. Due to the high xylan content in wheat straw, fermentation of both xylose and glucose is crucial to meet desired overall yields of ethanol. In the present work a recombinant xylose fermenting strain of Saccharomyces cerevisiae, TMB3400, cultivated aerobically on wheat straw hydrolysate, was used in simultaneous saccharification and fermentation (SSF) of steam pretreated wheat straw. The influence of fermentation strategy and temperature was studied in relation to xylose consumption, ethanol formation and by-product formation. In addition, model SSF experiments were made to further investigate the influence of temperature on xylose fermentation and by-product formation. In particular for SSF at the highest value of fibre content tested (9% water insoluble substance, WIS), it was found that a fed-batch strategy was clearly superior to the batch process in terms of ethanol yield, where the fed-batch gave 71% of the theoretical yield (based on all available sugars) in comparison to merely 59% for the batch. Higher ethanol yields, close to 80%, were obtained at a WIS-content of 7%. Xylose fermentation significantly contributed to the overall ethanol yields. The choice of temperature in the range 30-37 degrees C was found to be important, especially at higher contents of water insoluble solids (WIS). The optimum temperature was found to be 34 degrees C for the raw material and yeast strain studied. Model SSF experiments with defined medium showed strong temperature effects on the xylose uptake rate and xylitol yield.

Effect of reduction in yeast and enzyme concentrations in a simultaneous-saccharification-and-fermentation-based bioethanol process - Technical and economic evaluation

The ethanol production cost in a simultaneous saccharification and fermentation-based bioethanol process is influenced by the requirements for yeast production and for enzymes. The main objective of this study was to evaluate—technically and economically—the influence of these two factors on the production cost. A base case with 5 g/L of baker’s yeast and an initial concentration of water-insoluble solids of 5% resulted in an experimental yield of 85%. When these data were implemented in Aspen Plus, yeast was assumed to be produced from sugars in the hydrolysate, reducing the overall ethanol yield to 69%. The ethanol production cost was 4.80 SEK/L (2.34 US

Effect of reduction in yeast and enzyme concentrations in a simultaneous-saccharification-and-fermentation-based bioethanol process

/gal). When adapted yeast was used at 2 g/L, an experimental yield of 74% was achieved and the estimated ethanol production cost was the same as in the base case. A 50% reduction in enzyme addition resulted in an increased production cost, to 5.06 SEK/L (2.47 US

Controlled Fed-Batch Fermentations of Dilute-Acid Hydrolysate in Pilot Development Unit Scale

/gal) owing to reduced ethanol yield.

Controlled pilot development unit-scale fed-batch cultivation of yeast on spruce hydrolysates.

The ethanol production cost in a simultaneous saccharification and fermentation-based bioethanol process is influenced by the requirements for yeast production and for enzymes. The main objective of this study was to evaluate--technically and economically--the influence of these two factors on the production cost. A base case with 5 g/L of bakers yeast and an initial concentration of water-insoluble solids of 5% resulted in an experimental yield of 85%. When these data were implemented in Aspen Plus, yeast was assumed to be produced from sugars in the hydrolysate, reducing the overall ethanol yield to 69%. The ethanol production cost was 4.80 SEK/L (2.34 US

Fuel ethanol production from steam-pretreated corn stover using SSF at higher dry matter content

/gal). When adapted yeast was used at 2 g/L, an experimental yield of 74% was achieved and the estimated ethanol production cost was the same as in the base case. A 50% reduction in enzyme addition resulted in an increased production cost, to 5.06 SEK/L (2.47 US

A comparison between batch and fed-batch simultaneous saccharification and fermentation of steam pretreated spruce

/gal) owing to reduced ethanol yield.

Simultaneous saccharification and fermentation of steam-pretreated bagasse using Saccharomyces cerevisiae TMB3400 and Pichia stipitis CBS6054

Inhibitors formed during wood hydrolysis constitute a major problem in fermenting dilute-acid hydrolysates. By applying a fed-batch technique, the levels of inhibitory compounds may be held low, enabling high ethanol productivity. In this study, a previously developed fed-batch strategy was modified and implemented for use in pilot development unit (PDU) scale. The rate of total gas formation, measured with a mass flow meter, was used as input variable in the control algorithm. The feed rate in the PDU-scale experiments could be properly controlled based on the gas evolution from the reactor. In fed-batch experiments utilizing TMB 3000, an inhibitor-tolerant strain of Saccharomyces cerevisiae, close to 100% of the hexoses in the hydrolysate was converted.

Influence of strain and cultivation procedure on the performance of simultaneous saccharification and fermentation of steam pretreated spruce

Yeast production on hydrolysate is a likely process solution in large‐scale ethanol production from lignocellulose. The hydrolysate will be available on site, and the yeast has furthermore been shown to acquire an increased inhibitor tolerance when cultivated on hydrolysate. However, due to over‐flow metabolism and inhibition, efficient yeast production on hydrolysate can only be achieved by well‐controlled substrate addition. In the present work, a method was developed for controlled addition of hydrolysate to PDU (process development unit)‐scale aerobic fed‐batch cultivations of Saccharomyces cerevisiae TMB 3000. A feed rate control strategy, which maintains the ethanol concentration at a low constant level, was adapted to process‐like conditions. The ethanol concentration was obtained from on‐line measurements of the ethanol mole fraction in the exhaust gas. A computer model of the system was developed to optimize control performance. Productivities, biomass yields, and byproduct formation were evaluated. The feed rate control worked satisfactorily and maintained the ethanol concentration close to the setpoint during the cultivations. Biomass yields of 0.45 g/g were obtained on added hexoses during cultivation on hydrolysate and of 0.49 g/g during cultivation on a synthetic medium with glucose as the carbon source. Exponential growth was achieved with a specific growth rate of 0.18 h−1 during cultivation on hydrolysate and 0.22 h−1 during cultivation on glucose.