Project description:First-generation biofuel biorefineries may be a starting point for the development of new value chains, as their by-products and side streams retain nutrients and valuable molecules that may be recovered and valorized for high-value applications. This study provides a chemical characterization of post-fermentation corn oil and thin stillage, side streams of dry-grind corn bioethanol production, in view of their valorization. An overall long-term study was conducted on the two co-products collected over 1 year from a bioethanol plant. Water content, acid value, sedimentation, mineral composition, and fatty acid profiles were analyzed on post-fermentation corn oil. Results highlighted that its acid value was high (19.72-24.29 mg KOH/g), indicating high levels of free fatty acids, but stable over the year due to standardized operating conditions. The fatty acid profile was that typical of corn oil, with a prevalence of linoleic (54-59% of total fatty acids) over oleic (23-27%) and palmitic (12-17%) acids. Macronutrients, fatty acid, and mineral profiles were investigated in thin stillage. Results revealed the acidic pH (4.05-4.68) and high dilution (90-93% water) of this side stream. The dry mass was composed of fats (19-30%), proteins (8.8-12.8%), ash (8.7-9.5%), and fiber (7.3-9.8%). The concomitant presence of a variegate complex of molecules of nutritional interest in corn bioethanol co-products, with several potential high-value market applications, make the perspective of their recovery a promising strategy to create new cross-sector interconnections according to circular economy principles.
Project description:The economy of biorefineries is influenced not only by biofuel production from carbohydrates but also by the production of valuable compounds from largely underutilized industrial residues. Currently, the demand for many chemicals that could be made in a biorefinery, such as succinic acid (SA), medium-chain fatty acids (MCFAs), and lactic acid (LA), is fulfilled using petroleum, palm oil, or pure carbohydrates as raw materials, respectively. Thin stillage (TS), the residual liquid material following distillation of ethanol, is an underutilized coproduct from the starch biofuel industry. This carbon-rich material has the potential for chemical upgrading by microorganisms. Here, we explored the formation of different fermentation products by microbial communities grown on TS using different bioreactor conditions. At the baseline operational condition (6-day retention time, pH 5.5, 35°C), we observed a mixture of MCFAs as the principal fermentation products. Operation of a bioreactor with a 1-day retention time induced an increase in SA production, and a temperature increase to 55°C resulted in the accumulation of lactic and propionic acids. In addition, a reactor operated with a 1-day retention time at 55°C conditions resulted in LA accumulation as the main fermentation product. The prominent members of the microbial community in each reactor were assessed by 16S rRNA gene amplicon sequencing and phylogenetic analysis. Under all operating conditions, members of the Lactobacillaceae family within Firmicutes and the Acetobacteraceae family within Proteobacteria were ubiquitous. Members of the Prevotellaceae family within Bacteroidetes and Lachnospiraceae family within the Clostridiales order of Firmicutes were mostly abundant at 35°C and not abundant in the microbial communities of the TS reactors incubated at 55°C. The ability to adjust bioreactor operating conditions to select for microbial communities with different fermentation product profiles offers new strategies to explore and compare potentially valuable fermentation products from TS and allows industries the flexibility to adapt and switch chemical production based on market prices and demands.
Project description:Lactobacillus panis PM1 has the ability to produce 1,3-propanediol (1,3-PDO) from thin stillage (TS), which is the major waste material after bioethanol production, and is therefore of significance. However, the fact that L. panis PM1 cannot use glycerol as a sole carbon source presents a considerable problem in terms of utilization of this strain in a wide range of industrial applications. Accordingly, L. panis PM1 was genetically engineered to directly utilize TS as a fermentable substrate for the production of valuable platform chemicals without the need for exogenous nutrient supplementation (e.g., sugars and nitrogen sources). An artificial glycerol-oxidative pathway, comprised of glycerol facilitator, glycerol kinase, glycerol 3-phosphate dehydrogenase, triosephosphate isomerase, and NADPH-dependent aldehyde reductase genes of Escherichia coli, was introduced into L. panis PM1 in order to directly utilize glycerol for the production of energy for growth and value-added chemicals. A pH 6.5 culture converted glycerol to mainly lactic acid (85.43 mM), whereas a significant amount of 1,3-propanediol (59.96 mM) was formed at pH 7.5. Regardless of the pH, ethanol (82.16 to 83.22 mM) was produced from TS fermentations, confirming that the artificial pathway metabolized glycerol for energy production and converted it into lactic acid or 1,3-PDO and ethanol in a pH-dependent manner. This study demonstrates the cost-effective conversion of TS to value-added chemicals by the engineered PM1 strain cultured under industrial conditions. Thus, application of this strain or these research findings can contribute to reduced costs of bioethanol production.
Project description:One of the key elements influencing the efficiency of cellulosic ethanol production is the effective pretreatment of lignocellulosic biomass. The aim of the study was to evaluate the effect of microwave-assisted pretreatment of wheat stillage in the presence of sodium cumene sulphonate (NaCS) hydrotrope used for the production of second-generation bioethanol. As a result of microwave pretreatment, the composition of the wheat stillage biomass changed significantly when compared with the raw material used, before treatment. Microwave-assisted pretreatment with NaCS effectively reduced the lignin content and hemicellulose, making cellulose the dominant component of biomass, which accounted for 42.91 ± 0.10%. In post pretreatment, changes in biomass composition were also visible on FTIR spectra. The peaks of functional groups and bonds characteristic of lignins (C-O vibration in the syringyl ring, asymmetric bending in CH3, and aromatic skeleton C-C stretching) decreased. The pretreatment of the analyzed lignocellulosic raw material with NaCS resulted in the complete conversion of glucose to ethanol after 48 h of the process, with yield (in relation to the theoretical one) of above 91%. The highest observed concentration of ethanol, 23.57 ± 0.10 g/L, indicated the high effectiveness of the method used for the pretreatment of wheat stillage that did not require additional nutrient supplementation.
Project description:Here, we report the metagenomes from five anaerobic bioreactors, operated under different conditions, that were fed carbohydrate-rich thin stillage from a corn starch ethanol plant. The putative functions of the abundant taxa identified here will inform future studies of microbial communities involved in valorizing this and other low-value agroindustrial residues.
Project description:BackgroundBacteria may compete with yeast for nutrients during bioethanol production process, potentially causing economic losses. This is the first study aiming at the quantification and identification of Lactic Acid Bacteria (LAB) present in the bioethanol industrial processes in different distilleries of Brazil.ResultsA total of 489 LAB isolates were obtained from four distilleries in 2007 and 2008. The abundance of LAB in the fermentation tanks varied between 6.0 × 105 and 8.9 × 108 CFUs/mL. Crude sugar cane juice contained 7.4 × 107 to 6.0 × 108 LAB CFUs. Most of the LAB isolates belonged to the genus Lactobacillus according to rRNA operon enzyme restriction profiles. A variety of Lactobacillus species occurred throughout the bioethanol process, but the most frequently found species towards the end of the harvest season were L. fermentum and L. vini. The different rep-PCR patterns indicate the co-occurrence of distinct populations of the species L. fermentum and L. vini, suggesting a great intraspecific diversity. Representative isolates of both species had the ability to grow in medium containing up to 10% ethanol, suggesting selection of ethanol tolerant bacteria throughout the process.ConclusionsThis study served as a first survey of the LAB diversity in the bioethanol process in Brazil. The abundance and diversity of LAB suggest that they have a significant impact in the bioethanol process.
Project description:Phytate is the main form of phosphorus in corn ethanol coproducts and poses digestion issues in monogastric-animal feed. Extracting phytate as a commodity chemical will bring extra revenue to the corn ethanol industry and reduces potential phosphorus pollution from livestock waste management. We assessed a simplified scale-up approach of an ion-exchange separation system applied to extract phytate from thin stillage using volumetric parameters and simplifications of the van Deemter model. Thin stillage is one of the main byproducts generated on dry-grind corn-to-ethanol plants and accounts for the liquid portion of the bottom product generated in the ethanol distillation process. Thin stillage is rich in dissolved phytate, which served as the basis for an ion-exchange extraction system developed with a scalability factor of 50. Under the evaluated conditions, similar breakthrough profiles were obtained when similar Péclet and Stanton numbers were maintained for the scales studied, demonstrating that a simple and straightforward scale-up can be attained if special attention is given to maintaining both parameters as the basis of calculations of the plate numbers of ion-exchange columns.
Project description:BackgroundIndustrial biotechnology will play an increasing role in creating a more sustainable global economy. For conventional aerobic bioprocesses supplying O2 can account for 15% of total production costs. Microbubbles (MBs) are micron-sized bubbles that are widely used in industry and medical imaging. Using a fluidic oscillator to generate energy-efficient MBs has the potential to decrease the costs associated with aeration. However, little is understood about the effect of MBs on microbial physiology. To address this gap, a laboratory-scale MB-based Saccharomyces cerevisiae Ethanol Red propagation-fermentation bioethanol process was developed and analysed.ResultsAeration with MBs increased O2 transfer to the propagation cultures. Titres and yields of bioethanol in subsequent anaerobic fermentations were comparable for MB-propagated and conventional, regular bubble (RB)-propagated yeast. However, transcript profiling showed significant changes in gene expression in the MB-propagated yeast compared to those propagated using RB. These changes included up-regulation of genes required for ergosterol biosynthesis. Ergosterol contributes to ethanol tolerance, and so the performance of MB-propagated yeast in fed-batch fermentations sparged with 1% O2 as either RBs or MBs were tested. The MB-sparged yeast retained higher levels of ergosteryl esters during the fermentation phase, but this did not result in enhanced viability or ethanol production compared to ungassed or RB-sparged fermentations.ConclusionsThe performance of yeast propagated using energy-efficient MB technology in bioethanol fermentations is comparable to that of those propagated conventionally. This should underpin the future development of MB-based commercial yeast propagation.
Project description:Biogas production from nitrogen-rich feedstock results in release of ammonia (NH3), causing inhibition of the microbial process. The reported threshold ammonia value for stable biogas production varies greatly between studies, probably because of differences in operating conditions. Moreover, it is often difficult to separate the effect of ammonia inhibition from that of organic loading rate (OLR), as these two factors are often interrelated. This study attempted to distinguish the effects of ammonia and OLR by analysis of two laboratory-scale biogas reactors operating with thin stillage and subjected to an increase in free ammonia (from 0.30 to 1.1 g L(-1)) either by addition of an external nitrogen source (urea) or by increasing the OLR (3.2-6.0 g volatile solids L(-1) d(-1)). The results showed that ammonia concentration was detrimental for process performance, with the threshold for stability in both processes identified as being about 1 g NH3-N L(-1), irrespective of OLR. Analysis of the methanogenic community showed limited differences between the two reactors on order level and a clear increase in the abundance of Methanomicrobiales, particularly Methanoculleus sp., in response to increasing ammonia concentration. Further comprehensive molecular analysis revealed that diverse Methanoculleus species dominated in the reactors at a given ammonia level at different OLR. The acetogenic community was clearly affected by both ammonia concentration and OLR, suggesting that the volatile fatty acid load in relation to the higher OLR was important for the dynamics of this community.
Project description:Recovery of bioactive compounds from distillery waste could be an option for valorizing this waste. This study investigated how the extraction of polyphenols (which have antioxidant activity) from distillery stillage was affected by solvent type and concentration, extraction time, and method of extraction (conventional solid-liquid extraction, CSLE; ultrasound-assisted extraction, UAE; microwave-assisted extraction, MAE). Although recovery was similar with UAE and MAE, 3 min MAE with 80% ethanol and 80% methanol produced the highest yields of total phenolic content (TPC), total flavonoid content (TFC) and phenolic acids. With CSLE, TPC was 2.1-1.8-times lower than with MAE and 1.7-1.4-times lower than with UAE. Increasing the solvent concentration to 100% significantly decreased recovery. Six phenolic acids were recovered (ferulic and p-coumaric acid predominated), which were present mainly in the free form. There was a significant positive correlation between antioxidant activity, as measured with three methods (one based on the hydrogen atom transfer and two based on single electron transfer mechanisms), and phenolic acid content. With MAE and UAE, polyphenols were recovered more efficiently, with 2.1 times and 1.5 times higher antioxidant activity, and with 15 times and 9 times shorter extraction times, respectively, than with CSLE; thus, they can be considered "green" alternatives to CSLE.