Biomethane production and microbial strategies corresponding to high organic loading treatment for molasses wastewater in an upflow anaerobic filter reactor.

Molasses wastewater contains high levels of organic compounds, cations, and anions, causing operational problems for anaerobic biological treatment. In this study, an upflow anaerobic filter (UAF) reactor was employed to establish a high organic loading treatment system for molasses wastewater and further investigated the microbial community dynamics in response to this stressful operation. The biogas production increased with an increase in total organic carbon (TOC) loading rate from 1.0 to 14 g/L/day, and then it decreased with further TOC loading rate addition until 16 g/L/day. The UAF reactor achieved a maximum biogas production of 6800 mL/L/day with a TOC removal efficiency of 66.5% at a TOC loading rate of 14 g/L/day. Further microbial analyses revealed that both the bacterial and archaeal communities developed multiple strategies to maintain stable operation of the reactor at high organic loading (e.g., Proteiniphilum and Defluviitoga maintained high abundances throughout the operation; Tissierella temporarily dominated the bacterial community at TOC loading rates of 8.0 to 14 g/L/day; and multi-trophic Methanosarcina shifted as the dominant methanogen at the TOC loading rates of 8.0 to 16 g/L/day). This study presents insights into a high organic loading molasses wastewater treatment system and the microbial flexibility in methane fermentation in response to process disturbances.

Effect of distillery sewage sludge addition on performance and bacterial community dynamics during distilled grain waste composting.

This study evaluated the dynamics of physicochemical characteristics and bacterial communities during the co-composting of distilled grain waste (DGW) and distillery sewage sludge (SS), with DGW mono-composting as a control. Results showed that co-composting with SS significantly improved DGW degradation efficiency (61.38% vs. 54.13%) and end-product quality (seed germination index: 129.82% vs. 113.61%; N + P2O5 + K2O: 9.08% vs. 5.28%), compared to DGW mono-composting. Microbial community analysis revealed that co-composting accelerated the bacterial community succession rate and enhanced the abundance of the phyla Proteobacteria, Firmicutes, Chloroflexi, and Deinococcota by 45.86%, 4.38%, 37.49%, and 15.29%, respectively. Network analysis showed that DGW-SS co-composting altered the interactions among the bacterial genera and improved bacterial community stability. Spearman correlation analysis indicated that the correlation between bacterial genera and environmental factors was more significant in DGW-SS co-composting. Therefore, co-composting of DGW and SS is a suitable strategy for the treatment of solid byproducts from spirit distilleries.

Bacterial Community Structure and Metabolic Function Succession During the Composting of Distilled Grain Waste.

Distilled grain waste (DGW) can be converted to organic fertilizer via aerobic composting process without inoculating exogenous microorganisms. To illustrate the material conversion mechanism, this study investigated the dynamic changes of bacterial community structure and metabolic function involved in DGW composting. Results showed that a significant increase in microbial community alpha diversity was observed during DGW composting. Moreover, unique community structures occurred at each composting stage. The dominant phyla were Firmicutes, Proteobacteria, Actinobacteriota, Bacteroidota, Myxococcota, and Chloroflexi, whose abundance varied according to different composting stages. Keystone microbes can be selected as biomarkers for each stage, and Microbispora, Chryseolinea, Steroidobacter, Truepera, and Luteimonas indicating compost maturity. Co-occurrence network analysis revealed a significant relationship between keystone microbes and environmental factors. The carbohydrate and amino acid metabolism were confirmed as the primary metabolic pathways by metabolic function profiles. Furthermore, nitrogen metabolism pathway analysis indicated that denitrification and NH3 volatilization induced higher nitrogen loss during DGW composting. This study can provide new understanding of the microbiota for organic matter and nitrogen conversion in the composting process of DGW.

Biochar addition reduces nitrogen loss and accelerates composting process by affecting the core microbial community during distilled grain waste composting.

This study evaluated the impact of biochar addition on nitrogen (N) loss and the process period during distilled grain waste (DGW) composting. Results from the five treatments (0, 5, 10, 15, and 20% biochar addition) indicated that 10% biochar addition (DB10) was optimal, resulting in the lowest N loss, 25.69% vs. 40.01% in the control treatment. Moreover, the DGW composting period was shortened by approximately 14 days by biochar addition. The composition of the microbial community was not significantly altered with biochar addition in each phase, however, it did accelerate the microbial succession during DGW composting. N metabolism pathway prediction revealed that biochar addition enhanced nitrification and inhibited denitrification, and the latter phenomenon was the main reason for reducing N loss during DGW composting. Based on the above results, a potential mechanism model for biochar addition to reduce N loss during the DGW composting process was established.

Potential for reduced water consumption in biorefining of lignocellulosic biomass to bioethanol and biogas.

Increasing ethanol demand and public concerns about environmental protection promote the production of lignocellulosic bioethanol. Compared to that of starch- and sugar-based bioethanol production, the production of lignocellulosic bioethanol is water-intensive. A large amount of water is consumed during pretreatment, detoxification, saccharification, and fermentation. Water is a limited resource, and very high water consumption limits the industrial production of lignocellulosic bioethanol and decreases its environmental feasibility. In this review, we focused on the potential for reducing water consumption during the production of lignocellulosic bioethanol by performing pretreatment and fermentation at high solid loading, omitting water washing after pretreatment, and recycling wastewater by integrating bioethanol production and anaerobic digestion. In addition, the feasibility of these approaches and their research progress were discussed. This comprehensive review is expected to draw attention to water competition between bioethanol production and human use.

Production of bioethanol from unwashed-pretreated rapeseed straw at high solid loading.

Reduction in water consumption and increase in ethanol concentration are two main challenges for bioethanol production from lignocellulosic materials. To address the two challenges, the aim of this work was to study the production of bioethanol from unwashed-pretreated rapeseed straw (RS) at high solid loading. RS pretreated with 1% (w w-1) H2SO4 at 160 °C for 10 min resulted in excellent digestibility and fermentability of pretreated RS. The unwashed-pretreated RS was subjected to presaccharification and fed-batch simultaneous saccharification and fermentation (P-FB-SSF) at a final solid loading of 22% (w w-1). Ethanol concentration and ethanol yield of 53.1 g L-1 (equivalent to 4.1% (w w-1) based on fermentation slurry) and 72.4% were obtained, respectively. In total, 92.1 g water g-1 ethanol was consumed, a much smaller amount than that observed with washing after pretreatment or fermentation performed at lower solid loading.

Acclimation Improves Methane Production from Molasses Wastewater with High Salinity in an Upflow Anaerobic Filter Reactor: Performance and Microbial Community Dynamics.

This study evaluated the performance of an upflow anaerobic filter (UAF) reactor in the thermophilic methane fermentation of hypersaline molasses wastewater. The high salinity (~ 45 mS/cm) of the undiluted wastewater completely inhibited the biogas production. An acclimation strategy involving gradient dilution of the molasses wastewater was implemented to gradually increase the salt stress. Consequently, the biogas production was recovered, inhibited only slightly by the high salinity of the undiluted wastewater. The reactor steadily achieved a high total organic carbon (TOC) loading rate of 5 g/L/day, with approximately 60% TOC removal efficiency. Acclimation to the gradually increased salt stress leads to a relative abundance of some halotolerant microbes, such as bacteria from Arcobacter, Tissierella, and Ruminococcaceae, which increased as their hydrolytic and acidogenic abilities adjusted to the incremental increase in salinity. Additionally, hydrogenotrophic methanogens, especially Methanoculleus, showed greater resistance to hypersalinity than aceticlastic methanogens. These results suggest that acclimation of the fermentation microbial community to hypersalinity is an effective strategy to improve methane production from hypersaline molasses wastewater in thermophilic UAF reactors.

Insight into the microbiology of nitrogen cycle in the dairy manure composting process revealed by combining high-throughput sequencing and quantitative PCR.

Nitrogen cycling during composting process is not yet fully understood. This study explored the key genes involved in nitrogen cycling during dairy manure composting process using high-throughput sequencing and quantitative PCR technologies. Results showed that nitrogen fixation occurred mainly during the thermophilic and cooling phases, and significantly enhanced the nitrogen content of compost. Thermoclostridium stercorarium was the main diazotroph. Ammonia oxidation occurred during the maturation phase and Nitrosomonas sp. was the most abundant ammonia oxidizing bacteria. Denitrification contributed to the greatest nitrogen loss during the composting process. The nirK community was dominated by Luteimonas sp. and Achromobacter sp., while the nirS community was dominated by Alcaligenes faecalis and Pseudomonas stutzeri. The nosZ community varied in a succession of Halomonas ilicicola, Pseudomonas flexibili and Labrenzia alba dominated communities according to different composting phases. Based on these results, nitrogen cycling models for different phases of the dairy manure composting process were established.

Minimizing ammonia emissions from dairy manure composting by biofiltration using a pre-composted material as the packing media.

Compost-based biofiltration is a method widely used to mitigate ammonia emissions during composting. To improve the efficiency of a composting-biofiltration system, it is necessary to determine the most effective degree of composting at which to process the packing media used in the biofiltration system. In this study, materials pre-composted for 20 and 30 d (C20 and C30, respectively), and mature compost (CM) that had been treated for 60 d, were applied as biofilter media to remove ammonia from dairy manure composting exhaust gases. A comparison of the results revealed that the C30 biofilter not only completely removed ammonia, but also produced the least nitrogen loss (1.84%). The C20 biofilter exhibited an inferior performance, indicating that enough pre-composted time is necessary for material used as the packing media. Though the CM biofilter displayed good performance with regard to ammonia removal (97.8%), it had a high nitrogen loss (6.46%). A spearman rank correlation matrix revealed that the abundance of nitrogen cycle genes including amoA, nosZ, nirK, and nirS, had a strong correlation with the physicochemical properties such as nitrate content, carbon source, moisture content, and pH of the biofilter media. C30 provided advantageous conditions and contained a relatively high abundance of nitrifiers and the lowest abundance of denitrifiers. As a result, C30 rather than CM was a more appropriate biofilter media for ammonia removal. Moreover, the occurrence of biological nitrification during the dairy manure composting process indicates the effectiveness of a material for use as biofilter media.

Effects of ammonium and/or sulfide on methane production from acetate or propionate using biochemical methane potential tests.

The inhibitory effects of ammonium and sulfide on the methane production using acetate or propionate as a carbon source were investigated under different pH and temperature conditions. The methane production rate, duration of the lag phase, and inhibition threshold limit during methane production were estimated using the Gompertz equation and inhibitor mathematical model. The methane production rates at 53°C were 2.3-2.7 times higher than those at 35°C in the case of non-inhibitors. Increasing the NH4+ and/or S2- concentration decreased the methane production rate and increased the duration of the lag phase. For methane fermentation that was not acclimated to high NH4+ concentration, the critical NH4+ concentration beyond which methane fermentation would stop was 4000-5650 mg/L, depending on the pH, temperature, and carbon source. When NH4+ and S2- were coexistent, the critical NH4+ concentration decreased to approximately 3800 mg/L when propionate was used and to approximately 4450 mg/L when acetate was used. However, no synergistic effect of NH4+ and S2- on the methane production rate was found at an NH4+ concentration of < 5000 mg/L and S2- concentration of 50 mg/L.

Effect of organic loading rate on thermophilic methane fermentation of stillage eluted from ethanol fermentation of waste paper and kitchen waste.

Thermophilic methane fermentation was a valid approach for handling the stillage eluted from ethanol fermentation of waste paper and kitchen waste. The wide organic loading rate (OLR) range (2-14 g VTS/(L d)) for stable performance and relatively high energy recovery efficiency (79.0%) were achieved, and OLR of 8 g VTS/(L d) was optimum for achievement of highest biogas evolution and VTS removal efficiency. Microbial community analysis revealed that hydrolysis of cellulose was the critical step for methane production from the stillage.

Bioethanol from sugarcane bagasse: Focused on optimum of lignin content and reduction of enzyme addition.

To investigate the effect of delignification on enzymatic saccharification and ethanol fermentation of sugarcane bagasse (SCB), NaClO, NaOH, and Na2CO3 were used to prepare SCB with different lignin contents. We found that a lignin content of approximately 11% was sufficient for enzymatic saccharification and fermentation. Based on this result, an economical delignification pretreatment method using a combination of acid and alkali (CAA) was applied. Lignin content of 11.7% was obtained after CAA pretreatment with 0.5% w/v H2SO4 at 140 °C for 10 min and 1.0% w/v NaOH at 90 °C for 60 min. Presaccharification-simultaneous saccharification and fermentation (P-SSF) of the CAA-pretreated SCB resulted in an ethanol concentration of 43.8 g/L and an ethanol yield of 81.7%, with an enzyme loading of 15 FPU/g-CAA-pretreated SCB. Enzyme activities (filter paper, carboxymethyl cellulase, and ß-glucosidase activities) were determined in liquid phase during P-SSF, indicating that the residual cellulase activity could be further used. Thus, fed-batch P-SSF was carried out, and an ethanol concentration of 43.1 g/L and an ethanol yield of 80.4% were obtained with an enzyme loading of 10 FPU/g-CAA-pretreated SCB. Fed-batch P-SSF was found to be effective to reduce enzyme loading.

Biogas Production from Distilled Grain Waste by Thermophilic Dry Anaerobic Digestion: Pretreatment of Feedstock and Dynamics of Microbial Community.

Distilled grain waste (DGW) eluted from the Chinese liquor making process poses potential serious environmental problems. The objective of this study is to evaluate the feasibility of converting DGW to biogas by thermophilic dry anaerobic digestion. To improve biogas production, the effects of dilute H2SO4 and thermal pretreatment on DGW were evaluated by biochemical methane potential (BMP) tests. The results indicate that 90 °C thermal pretreatment provided the highest methane production at 212.7 mL/g-VTSadd. The long-term thermophilic dry anaerobic digestion process was conducted in a 5-L separable flask for more than 3 years at a volatile total solid (VTS) loading rate of 1 g/kg-sludge/d, using synthetic waste, untreated and 90 °C thermal pretreated DGW as the feedstock, respectively. A higher methane production, 451.6 mL/g-VTSadd, was obtained when synthetic waste was used; the methane production decreased to 139.4 mL/g-VTSadd when the untreated DGW was used. The 90 °C thermal pretreated DGW increased the methane production to 190.5 mL/g-VTSadd, showing an increase of 36.7% in methane production compared with that using untreated DGW. The microbial community structure analysis indicates that the microbial community in the thermophilic dry anaerobic digestion system maintained a similar structure when untreated or pretreated DGW was used, whereas the structure differed significantly when synthetic waste was used as the feedstock.

A comparative study of composting the solid fraction of dairy manure with or without bulking material: Performance and microbial community dynamics.

The present study compared the development of various physicochemical properties and the composition of microbial communities involved in the composting process in the solid fraction of dairy manure (SFDM) with a sawdust-regulated SFDM (RDM). The changes in several primary physicochemical properties were similar in the two composting processes, and both resulted in mature end-products within 48days. The bacterial communities in both composting processes primarily comprised Proteobacteria and Bacteroidetes. Firmicutes were predominant in the thermophilic phase, whereas Chloroflexi, Planctomycetes, and Nitrospirae were more abundant in the final mature phase. Furthermore, the succession of bacteria in both groups proceeded in a similar pattern, suggesting that the effects of the bulking material on bacterial dynamics were minor. These results demonstrate the feasibility of composting using only the SFDM, reflected by the evolution of physicochemical properties and the microbial communities involved in the composting process.

Production of Bio-Ethanol by Integrating Microwave-Assisted Dilute Sulfuric Acid Pretreated Sugarcane Bagasse Slurry with Molasses.

Sugarcane bagasse (SCB) and molasses, known as carbohydrate-rich biomass derived from sugar production, can serve as feedstock for bio-ethanol production. To establish a simple process, the production of bio-ethanol through integration of whole pretreated slurry (WPS) of SCB with molasses was investigated. The results showed that microwave-assisted dilute sulfuric acid pretreatment reduced the formation of toxic compounds compared to a pretreatment process involving "conventional heating". Pretreatment at 180 oC with 10% w v-1 solid loading and 0.5% w v-1 H2SO4 was sufficient to achieve efficient enzymatic saccharification of WPS. By conducting separate hydrolysis and fermentation (SHF), an ethanol yield of 90.12% was obtained from the mixture of WPS and molasses, but the ethanol concentration of 33.48 g L-1 was relatively low. By adopting fed-batch SHF, the ethanol concentration reached 41.49 g L-1. Assuming that the molasses were converted to ethanol at an efficiency of 87.21% (i.e., ethanol was obtained from fermentation of molasses alone), the ethanol yield from WPS when a mixture of WPS and molasses was fermented was 78.30%, which was higher than that of enzymatic saccharification of WPS (73.53%). These findings suggest that the production of bio-ethanol via integration of WPS with molasses is a superior method. Graphical Abstract ᅟ.

Inhibitor tolerance of a recombinant flocculating industrial Saccharomyces cerevisiae strain during glucose and xylose co-fermentation

ABSTRACT Lignocellulose-derived inhibitors have negative effects on the ethanol fermentation capacity of Saccharomyces cerevisiae. In this study, the effects of eight typical inhibitors, including weak acids, furans, and phenols, on glucose and xylose co-fermentation of the recombinant xylose-fermenting flocculating industrial S. cerevisiae strain NAPX37 were evaluated by batch fermentation. Inhibition on glucose fermentation, not that on xylose fermentation, correlated with delayed cell growth. The weak acids and the phenols showed additive effects. The effect of inhibitors on glucose fermentation was as follows (from strongest to weakest): vanillin > phenol > syringaldehyde > 5-HMF > furfural > levulinic acid > acetic acid > formic acid. The effect of inhibitors on xylose fermentation was as follows (from strongest to weakest): phenol > vanillin > syringaldehyde > furfural > 5-HMF > formic acid > levulinic acid > acetic acid. The NAPX37 strain showed substantial tolerance to typical inhibitors and showed good fermentation characteristics, when a medium with inhibitor cocktail or rape straw hydrolysate was used. This research provides important clues for inhibitors tolerance of recombinant industrial xylose-fermenting S. cerevisiae.

Aerobic composting of distilled grain waste eluted from a Chinese spirit-making process: The effects of initial pH adjustment.

Aerobic composting of distilled grain waste (DGW) at different initial pH values adjusted by CaO addition was investigated. Three pH-adjusted treatments with initial pH values of 4 (R1), 5 (R2) and 6 (R3) and a control treatment (R0) with a pH value of 3.5 were conducted simultaneously. The results showed that R0 had an unsuccessful start-up of composting. However, the pH-adjusted treatments produced remarkable results, with a relatively high initial pH being beneficial for the start-up. Within 65days of composting, the degradation of volatile solids (VS) and the physicochemical properties of R2 and R3 displayed similar tendencies, and both produced a mature end-product, while R1 exhibited a lower VS degradation rate, and some of its physicochemical properties indicated the end-product was immature. Quantitative PCR analysis of ammonia oxidizers indicated that the occurrence of nitrification during the composting of DGW could be attributed to the activity of ammonia-oxidizing bacteria.

Vinegar production from post-distillation slurry deriving from rice shochu production with the addition of caproic acid-producing bacteria consortium and lactic acid bacterium.

To establish a zero emission process, the post-distillation slurry of a new type of rice shochu (NTRS) was used for the production of health promoting vinegar. Since the NTRS post-distillation slurry contained caproic acid and lactic acid, the effect of these two organic acids on acetic acid fermentation was first evaluated. Based on these results, Acetobacter aceti CICC 21684 was selected as a suitable strain for subsequent production of vinegar. At the laboratory scale, acetic acid fermentation of the NTRS post-distillation slurry in batch mode resulted in an acetic acid concentration of 41.9 g/L, with an initial ethanol concentration of 40 g/L, and the acetic acid concentration was improved to 44.5 g/L in fed-batch mode. Compared to the NTRS post-distillation slurry, the vinegar product had higher concentrations of free amino acids and inhibition of angiotensin I converting enzyme activity. By controlling the volumetric oxygen transfer coefficient to be similar to that of the laboratory scale production, 45 g/L of acetic acid was obtained at the pilot scale, using a 75-L fermentor with a working volume of 40 L, indicating that vinegar production can be successfully scaled up.

Inhibitor tolerance of a recombinant flocculating industrial Saccharomyces cerevisiae strain during glucose and xylose co-fermentation.

Lignocellulose-derived inhibitors have negative effects on the ethanol fermentation capacity of Saccharomyces cerevisiae. In this study, the effects of eight typical inhibitors, including weak acids, furans, and phenols, on glucose and xylose co-fermentation of the recombinant xylose-fermenting flocculating industrial S. cerevisiae strain NAPX37 were evaluated by batch fermentation. Inhibition on glucose fermentation, not that on xylose fermentation, correlated with delayed cell growth. The weak acids and the phenols showed additive effects. The effect of inhibitors on glucose fermentation was as follows (from strongest to weakest): vanillin>phenol>syringaldehyde>5-HMF>furfural>levulinic acid>acetic acid>formic acid. The effect of inhibitors on xylose fermentation was as follows (from strongest to weakest): phenol>vanillin>syringaldehyde>furfural>5-HMF>formic acid>levulinic acid>acetic acid. The NAPX37 strain showed substantial tolerance to typical inhibitors and showed good fermentation characteristics, when a medium with inhibitor cocktail or rape straw hydrolysate was used. This research provides important clues for inhibitors tolerance of recombinant industrial xylose-fermenting S. cerevisiae.

Production of ethanol from a mixture of waste paper and kitchen waste via a process of successive liquefaction, presaccharification, and simultaneous saccharification and fermentation.

Efficient ethanol production from waste paper requires the addition of expensive nutrients. To reduce the production cost of ethanol from waste paper, a study on how to produce ethanol efficiently by adding kitchen waste (potentially as a carbon source, nutrient source, and acidity regulator) to waste paper was performed and a process of successive liquefaction, presaccharification, and simultaneous saccharification and fermentation (L+PSSF) was developed. The individual saccharification performances of waste paper and kitchen waste were not influenced by their mixture. Liquefaction of kitchen waste at 90°C prior to presaccharification and simultaneous saccharification and fermentation (PSSF) was essential for efficient ethanol fermentation. Ethanol at concentrations of 46.6 or 43.6g/l was obtained at the laboratory scale after fermentation for 96h, even without pH adjustment and/or the addition of extra nutrients. Similarly, ethanol at a concentration of 45.5g/l was obtained at the pilot scale after fermentation for 48h. The ethanol concentration of L+PSSF of the mixture of waste paper and kitchen waste was comparable to that of PSSF of waste paper with added nutrients (yeast extract and peptone) and pH adjustment using H2SO4, indicating that kitchen waste is not only a carbon source but also an excellent nutrient source and acidity regulator for fermentation of the mixture of waste paper and kitchen waste.