Simultaneous Saccharification And Fermentation Mode Research Articles

Efficient biosynthesis of exopolysaccharide from Jerusalem artichoke using a novel strain of Bacillus velezensis LT-2

This study focused on the non-grain biorefining of Jerusalem artichoke (JA) for exopolysaccharide (EPS) efficient production by using Bacillus velezensis LT-2. Results showed that LT-2 could directly utilize JA tuber power (JATP) for EPS production, and its EPS yield reached 11.47 ± 0.33 g/L in the simultaneous saccharification and fermentation (SSF) mode. Furthermore, the SSF mode shortened the fermentation period by 26.67% and reduced the fermentation cost by 79.41% due to the improved substrate utilization and the avoidance of inhibition effects of a high fructose concentration. Transcriptome sequencing results showed that inulin could accelerate nucleotide-sugars biosynthesis, induce EPS synthetic gene cluster transcription, and strengthen the electron transport respiratory chain and the transporter systems, thereby ensuring EPS efficient synthesis. This work exhibited a productive non-grain and environmentally friendly fermentation strategy for EPS biosynthesis, which promoted the JA industry development and created new prospects for high-value industrial products biosynthesis by using JATP.

Designing Efficient Processes for Sustainable Bioethanol and Bio-Hydrogen Production from Grass Lawn Waste.

The effect of thermal, acid and alkali pretreatment methods on biological hydrogen (BHP) and bioethanol production (BP) from grass lawn (GL) waste was investigated, under different process schemes. BHP from the whole pretreatment slurry of GL was performed through mixed microbial cultures in simultaneous saccharification and fermentation (SSF) mode, while BP was carried out through the C5yeast Pichia stipitis, in SSF mode. From these experiments, the best pretreatment conditions were determined and the efficiencies for each process were assessed and compared, when using either the whole pretreatment slurry or the separated fractions (solid and liquid), the separate hydrolysis and fermentation (SHF) or SSF mode, and especially for BP, the use of other yeasts such as Pachysolen tannophilus or Saccharomyces cerevisiae. The experimental results showed that pretreatment with 10 gH2SO4/100 g total solids (TS) was the optimum for both BHP and BP. Separation of solid and liquid pretreated fractions led to the highest BHP (270.1 mL H2/g TS, corresponding to 3.4 MJ/kg TS) and also BP (108.8 mg ethanol/g TS, corresponding to 2.9 MJ/kg TS) yields. The latter was achieved by using P. stipitis for the fermentation of the hydrolysate and S. serevisiae for the solid fraction fermentation, at SSF.

Bioethanol Production from Alkali Steam Explosion of Oil Palm of Empty Fruit Bunch Fiber

Bioethanol is the most promising biofuels and can be generated from lignocellulosic biomasses. The study of bioethanol production from oil palm empty fruit bunch (OPEFB) pretreated by alkali steam explosion was carried out employing separate enzymatic hydrolysis (SHF) and Simultaneous Saccharification and Fermentation (SSF) approaches. Alkaline extraction after steam explosion pretreatment of OPEFB led to the partial removal of lignin, hemicellulose and other degradation products from fiber surface. The pretreatment process is of great importance to ethanol yield. In the present study, the pretreatment was processed using a steam explosion reactor at 150°C, 4 bars for 30 minute with alkaline concentrations of NaOH 10% (kg/L). After pretreatment, the OPEFB substrates 10% and 15% (w/v) was saccharification using 30 FPU of Ctec2 and Htec2 enzyme and fermentation using yeast of Saccharomyces cerevisiae for 72 hours. The glucose obtained after 72 hours was 137.80 g/L (subtrate 15%) and 97.90 g/L (subtrate 10%). Bioethanol production in SSF mode faster and slightly more efficient process than SHF.

Enzymatic Conversion of Sugar Beet Pulp: A Comparison of Simultaneous Saccharification and Fermentation and Separate Hydrolysis and Fermentation for Lactic Acid Production.

This study compares the efficiency of lactic acid production by separate hydrolysis and fermentation (SHF) or simultaneous saccharification and fermentation (SSF) of sugar beet pulp, a byproduct of industrial sugar production. In experiments, sugar beet pulp was hydrolyzed using five commercial enzymes. A series of shake flask fermentations were conducted using five selected strains of lactic acid bacteria (LAB). The differences in the activities of the enzymes for degrading the principal sugar beet pulp components were reflected in the different yields of total reducing sugars. The highest yields after hydrolysis and the lowest quantities of insoluble residues were obtained using a mixture (1:1) of Viscozyme® and Ultraflo® Max. In the SHF process, only a portion of the soluble sugars released by the enzymes from the sugar beet pulp was assimilated by the LAB strains. In SSF, low enzyme loads led to reduction in the efficiency of sugar accumulation. The risk of carbon catabolic repression was reduced. Our results suggest that SSF has advantages over SHF, including lower processing costs and higher productivity. Lactic acid yield in SSF mode (approx. 30 g/L) was 80-90% higher than that in SHF.

Production of poly(3-hydroxybutyrate) by simultaneous saccharification and fermentation of cereal mash using Halomonas boliviensis

The production of biodegradable polymer poly(3-hydroxybutyrate) (PHB) by the halophilic bacterium Halomonas boliviensis, using cereal mash as the carbon source, was carried out in this study. Simultaneous saccharification and fermentation (SSF) methodology, commonly used in first generation (1G) biorefineries, was applied in order to evaluate the integration of PHB production in a bioethanol plant. Separate hydrolysis and fermentation (SHF) was compared to the SSF process based on the results of overall mass balance and productivities. The highest PHB production (26gL⿿1) was obtained by applying the SSF configuration, which was 60% higher than the value reached with the SHF mode. Moreover, the application of the SSF mode permitted a 47% reduction in the overall processing time. This study demonstrates, for the first time, the feasibility of applying an SSF process for producing PHB from a real cereal mash and its integration within a 1G ethanol biorefinery.

Bioethanol production from steam explosion pretreated and alkali extracted Cistus ladanifer (rockrose)

Biofuels are suitable alternatives to conventional and fossil fuels that pose serious environmental adverse effects to society. Bioethanol is the most promising biofuel and can be generated from lignocellulosic biomasses. Forest residues are outside the human food chain and inexpensive raw materials, but they need a previous pretreatment, in order to improve the cellulose accessibility for further bioconversion.The study of bioethanol production from rockrose pretreated by steam explosion (SE) was carried out employing separate enzymatic hydrolysis and fermentation (SHF) or simultaneous saccharification and fermentation (SSF) approaches. Saccharification of untreated rockrose attained only 0.9% of sugars yield. However, the steam explosion pretreatment promoted the disruption of interfibrillar surfaces of fibers with partial degradation of lignin thus enhancing the accessibility of polysaccharides toward enzymatic hydrolysis. Alkaline extraction after steam explosion pretreatment of rockrose residue (R-SE-OH) led to the partial removal of lignin, hemicelluloses, and other degradation products from fibre surface allowing an increase of 75% in the glucose yield. Bioethanol production in SSF mode was faster and slightly more efficient process than SHF providing the best results: ethanol concentration of 16.1gL−1, fermentation efficiency of 69.8% and a yield of 22.1g ethanol/100g R-SE-OH.

Enzymatic saccharification and bioethanol production from Cynara cardunculus pretreated by steam explosion

The correct choice of the specific lignocellulosic biomass pretreatment allows obtaining high biomass conversions for biorefinery implementations and cellulosic bioethanol production from renewable resources. Cynara cardunculus (cardoon) pretreated by steam explosion (SE) was involved in second-generation bioethanol production using separate hydrolysis and fermentation (SHF) or simultaneous saccharification and fermentation (SSF) processes. Steam explosion pretreatment led to partial solubilisation of hemicelluloses and increased the accessibility of residual polysaccharides towards enzymatic hydrolysis revealing 64% of sugars yield against 11% from untreated plant material. Alkaline extraction after SE pretreatment of cardoon (CSEOH) promoted partial removal of degraded lignin, tannins, extractives and hemicelluloses thus allowing to double glucose concentration upon saccharification step. Bioethanol fermentation in SSF mode was faster than SHF process providing the best results: ethanol concentration 18.7gL−1, fermentation efficiency of 66.6% and a yield of 26.6g ethanol/100g CSEOH or 10.1g ethanol/100g untreated cardoon.

Attainable region analysis for continuous production of second generation bioethanol

BackgroundDespite its semi-commercial status, ethanol production from lignocellulosics presents many complexities not yet fully solved. Since the pretreatment stage has been recognized as a complex and yield-determining step, it has been extensively studied. However, economic success of the production process also requires optimization of the biochemical conversion stage. This work addresses the search of bioreactor configurations with improved residence times for continuous enzymatic saccharification and fermentation operations. Instead of analyzing each possible configuration through simulation, we apply graphical methods to optimize the residence time of reactor networks composed of steady-state reactors. Although this can be easily made for processes described by a single kinetic expression, reactions under analysis do not exhibit this feature. Hence, the attainable region method, able to handle multiple species and its reactions, was applied for continuous reactors. Additionally, the effects of the sugars contained in the pretreatment liquor over the enzymatic hydrolysis and simultaneous saccharification and fermentation (SSF) were assessed.ResultsWe obtained candidate attainable regions for separate enzymatic hydrolysis and fermentation (SHF) and SSF operations, both fed with pretreated corn stover. Results show that, despite the complexity of the reaction networks and underlying kinetics, the reactor networks that minimize the residence time can be constructed by using plug flow reactors and continuous stirred tank reactors. Regarding the effect of soluble solids in the feed stream to the reactor network, for SHF higher glucose concentration and yield are achieved for enzymatic hydrolysis with washed solids. Similarly, for SSF, higher yields and bioethanol titers are obtained using this substrate.ConclusionsIn this work, we demonstrated the capabilities of the attainable region analysis as a tool to assess the optimal reactor network with minimum residence time applied to the SHF and SSF operations for lignocellulosic ethanol production. The methodology can be readily modified to evaluate other kinetic models of different substrates, enzymes and microorganisms when available. From the obtained results, the most suitable reactor configuration considering residence time and rheological aspects is a continuous stirred tank reactor followed by a plug flow reactor (both in SSF mode) using washed solids as substrate.

Simultaneous Saccharification and Ethanol Fermentation of Corn Stover at High Temperature and High Solids Loading by a Thermotolerant Strain Saccharomyces cerevisiae DQ1

The thermotolerant strain Saccharomyces cerevisiae DQ1 was applied to the simultaneous saccharification and fermentation (SSF) at high temperature and high solids loading of the dilute acid-pretreated corn stover in the present study. The SSF using S. cerevisiae DQ1 was operated at 30 % solids loading of the pretreated corn stover with three-step SSF mode and achieved up to ethanol titer of 48 g/L and yield of 65.6 %. S. cerevisiae DQ1 showed strong thermotolerance in both the regular one-step SSF and the three-step SSF with changing temperature in each step. The three-step SSF at 40°C using S. cerevisiae DQ1 tolerated the greater cellulase dosage and solids loading of the pretreated corn stover and resulted in increased ethanol production. The present study provided a practical potential for the future SSF of lignocellulose feedstock at high temperature to reach high ethanol titer.

Comparative performance of precommercial cellulases hydrolyzing pretreated corn stover

BackgroundCellulases and related hydrolytic enzymes represent a key cost factor for biochemical conversion of cellulosic biomass feedstocks to sugars for biofuels and chemicals production. The US Department of Energy (DOE) is cost sharing projects to decrease the cost of enzymes for biomass saccharification. The performance of benchmark cellulase preparations produced by Danisco, DSM, Novozymes and Verenium to convert pretreated corn stover (PCS) cellulose to glucose was evaluated under common experimental conditions and is reported here in a non-attributed manner.ResultsTwo hydrolysis modes were examined, enzymatic hydrolysis (EH) of PCS whole slurry or washed PCS solids at pH 5 and 50°C, and simultaneous saccharification and fermentation (SSF) of washed PCS solids at pH 5 and 38°C. Enzymes were dosed on a total protein mass basis, with protein quantified using both the bicinchoninic acid (BCA) assay and the Bradford assay. Substantial differences were observed in absolute cellulose to glucose conversion performance levels under the conditions tested. Higher cellulose conversion yields were obtained using washed solids compared to whole slurry, and estimated enzyme protein dosages required to achieve a particular cellulose conversion to glucose yield were extremely dependent on the protein assay used. All four enzyme systems achieved glucose yields of 90% of theoretical or higher in SSF mode. Glucose yields were reduced in EH mode, with all enzymes achieving glucose yields of at least 85% of theoretical on washed PCS solids and 75% in PCS whole slurry. One of the enzyme systems ('enzyme B') exhibited the best overall performance. However in attaining high conversion yields at lower total enzyme protein loadings, the relative and rank ordered performance of the enzyme systems varied significantly depending upon which hydrolysis mode and protein assay were used as the basis for comparison.ConclusionsThis study provides extensive information about the performance of four precommercial cellulase preparations. Though test conditions were not necessarily optimal for some of the enzymes, all were able to effectively saccharify PCS cellulose. Large differences in the estimated enzyme dosage requirements depending on the assay used to measure protein concentration highlight the need for better consensus methods to quantify enzyme protein.