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Exploiting highly active and methanol-resistant lipase is of great significance for biodiesel production. A semi-rational directed evolution method combined with N-glycosylation is reported, and all mutants exhibiting higher catalytic activity and methanol tolerance than the wild type (WT). Mutant N267 retained 64% activity after incubation in 50% methanol for 8 h, which was 48% greater than that of WT. The catalytic activity of mutants N267 and N167 was 30- and 71- fold higher than that of WT. Molecular dynamics simulations of N267 showed that the formation of new strong hydrogen bonds between glycan and the protein stabilized the structure of lipase and improved its methanol tolerance. N267 achieved biodiesel yields of 99.33% (colza oil) and 81.70% (waste soybean oil) for 24 h, which was much higher than WT (51.6% for rapeseed oil and 44.73% for wasted soybean oil). The engineered ProRML mutant has high potential for commercial biodiesel production.

The long-term exploitation of ionic rare earth elements (REEs) in southern China has produced a large-scale of abandoned tailings area. While the application of anaerobic fermentation residues to cultivate economically valuable remediation plants (e.g. energy plant) has become a hotspot due to their merits in low-cost and sustainability in recent years, the succession and co-occurrence patterns of these microbial communities remain unclear. In this study, soil samples were collected from the sustainable restoration area, natural restoration area and tailings area. The composition and diversity of bacterial and fungal communities on five soil samples were evaluated using high-throughput sequencing technology. The results shown that the phytoremediation with anaerobic fermentation residues could significantly improve the physicochemical properties (especially for soil nutrients) and microbial diversity of soil within 3 years, while these parameters in natural restoration area were lower. The nonmetric multidimensional scaling (NMDS) ordinations revealed the shifts of microbial communities depending on soil physicochemical properties and plant species, and soil nutrients were the main factors affecting the microbial variation explained by the variation partition analysis (VPA). The soil nutrient accumulation obviously changed the proportion of oligotrophic and copiotrophic groups, among which the copiotrophic groups were significantly increased, such as Proteobacteria, Bacteroidetes, Gemmatimonadetes and Glomeromycota. The microbial co-occurrence network analysis indicated that application of anaerobic fermentation residues could significantly improve the topological properties and the stability of microbial network. The copiotrophic groups (e.g. Proteobacteria, Ascomycota) became the key to assemble stable network structure. Moreover, herbaceous plants could increase the proportion of fungi (e.g. Ascomycota) in microbial network, which improved the topological properties with bacteria synergistically. Therefore, the soil environment of REEs tailings area was effectively optimized by anaerobic fermentation residues and herbaceous plants, which furthered understanding of co-occurrence pattern and mutualistic relationships of microbial communities during sustainable restoration.

In this work, the impact of initial pH on the production of volatile fatty acids (VFAs) of hybrid Pennisetum was investigated. The batch experiments were conducted under six distinct beginning pH at a mesophilic temperature. Initial pH had an obvious effect on VFA yield and composition, and severe alkaline circumstances (pHin = 11.0) could boost VFA production and acetic acid selectivity. The highest VFAs yield and acetate proportion were obtained when the initial pH was 11.0, with 518 ± 29 mg g−1VS and 92%. Furthermore, microbial community analysis showed that alkaliphilic acetogenic anaerobe such as Amphibacillus, Tissierella, and Natronincola were the dominant species when the initial pH was 11.0. The Amphibacillus is the main hydrolysis bacterium under these conditions because of its high ability for xylan degradation at pH 9.0–10.0. Because of the increased VFA yield and superior acetic acid selectivity, the results suggest that adjusting the initial pH to 11.0 in batch mode would be possible for scaling-up purposes.

This study investigated whether bioaugmentation can improve the anaerobic digestion (AD) performance of food waste (FW), as well as the effects of addition dosage and frequency on the bioaugmentation’s performance and economic feasibility. The findings demonstrated that all the bioaugmented digesters, regardless of dosage and frequency, performed more effectively in biogas production than the non-bioaugmentation control. Furthermore, relatively higher dosages or frequencies increased AD performance. Introducing 0.25 g L−1 d−1 of bioaugmentation seed every three days increased OLR and volumetric biogas production 8-fold and 12-fold, respectively, compared to the non-bioaugmentation control. Whole-genome sequencing analysis showed that bioaugmentation enhanced the population of the acetoclastic Methanothrix (belong to the order Methanosarcinales). Moreover, high abundance of Methanothrix (exceeding 80%) contributed to a better AD performance. Economic analysis of an up-scale biogas plant suggested that an appropriate bioaugmentation process increased income, thus increasing the profit to 3696 CNY d−1 if treated at 21 t FW.

The anaerobic digestion (AD) performance for breakfast, lunch, and dinner kitchen waste (KW) produced by the canteen at Guangzhou University was assessed by a batch experiment under mesophilic and thermophilic conditions. The AD concentrations were 9.54 g VS/L, 20.12 g VS/L, and 39.99 g VS/L, respectively. The concentration of 20.12 g VS/L under thermophilic conditions had the best AD performance among all the KW. The maximum specific methane yield of 591 ± 30 mL/g VS was obtained from dinner KW (D-KW). Due to the differing organic composition, the specific methane yield of D-KW and lunch KW (L-KW) was 41.16% and 19.76% higher than that of breakfast KW (B-KW) under the optimal condition, respectively, whereas that of D-KW was 17.96% higher than that of L-KW. The specific methane yield was significantly correlated to the crude fat content. When the concentration was increased to 39.99 g VS/L, accumulation of toxic materials was observed and the AD process stopped. The potential biogas yield, based on production of 5904.43 tons of KW per year at the Guangzhou Higher Education Mega Center (HEMC), is 1.08 million m3 biogas, equivalent to 2.15 million KW h of electricity.

The acid-tolerant methanogenic propionate degradation culture was acclimated in a propionate-fed semi-continuous bioreactor by daily adjusting the digestate pH. The performance of propionate fermentation, the respond of microbial community structure to the acidic environment, and the microbial network for propionate degradation in the acid-tolerant culture was investigated. The results demonstrated that after long term of acclimation to low pH, the digester could produce methane from propionate at pH 4.8–5.5 with 0.3–0.4 L g−1 propionic acid (HPr) d−1 of the volatile solids (VS) methane production. The predominant methanogens shifted from acetoclastic methanogens (∼87%) to hydrogenotrophic methanogens (∼67%) in the bioreactor with the dropping pH, indicating that hydrogenotrophic methanogens were more acid-tolerant than acetoclastic methanogens. Smithella (∼11%), Syntrophobacter (∼7%) and Pelotomaculum (∼3%) were the main propionate oxidizers in the acid-tolerant propionate-utilizing culture. Methanothrix dominant acetoclastic methanogens, while Methanolinea and Methanospirillum were the major H2 scavengers to support Syntrophobacter and Pelotomaculum syntrophic propionate degradation.

The methanogenic propionate degradation consortia were enriched in a propionate-fed semi-continuous bioreactor. The microbial community shift with depth, the microbial network and its correlation with metabolic pathway were also investigated. The results demonstrated that the maximum organic loading rate (OLR) of the reactor was 2.5 g propionic acid (HPr) L−1 d−1 with approximately 1.20 LL−1 d−1 of volumetric methane production (VMP). The organisms in the enrichment were spanning 36 bacterial phyla and 7 archaeal orders. Syntrophobacter, the main Hpr oxidizer in the digester, dominated bacteria with relative abundance changing from 63% to 37% with depth. The predominant methanogens shift from hydrogenotrophic Methanoculleus (∼60%) at the upper liquid layer to acetoclastic Methanothrix (∼51%) at the lower sediment layer in the bioreactor. These methanogens syntrophically support Syntrophobacter by degrading HPr catabolism by-products (H2 and acetate). Other bacteria could scavenge anabolic products (carbohydrate and protein) presumably derived from detrital biomass produced by the HPr-degrading community.

This study is the first to integrate gamma-valerolactone/water (GVL/water) pretreatment with anaerobic digestion (AD) for biogas production and lignin nanoparticles (LNPs) synthesis. The hydrothermal treatment was conducted at 135 to 180 °C with GVL at 0 to 90%. After pretreatment, the compositions of hybrid pennisetum were changed with the removal of lignin, hemicellulose, and cellulose to different extent. Subsequent anaerobic digestion achieved a maximal specific methane yield of 228.00 ± 4.37 mL/g VS, compared with that at 165.11 ± 1.99 mL/g VS for the control. The highest actual methane yield (150 mL/g RM) was achieved by pretreatment with GVL/water (50/50) at 150 °C for 90 min. LNPs at 200 to 2000 nm were synthesized from the liquid waste with a yield at ~4 mg/mL. The mass balance of this integrated method was discussed. In general, the maximal valorisation of hybrid pennisetum was achieved based on a catalyst-free of GVL/water pretreatment.

To enhance the biodegradability and methane production of hybrid Pennisetum, a pretreatment method with high selectivity for lignin removal, namely sodium chlorite/acetic acid (SCA) pretreatment, was examined in this work. Results showed that SCA pretreatment can selectively remove lignin with minimal impact on cellulose and hemicellulose. After up to 200 min of SCA treatment, 79.4% of lignin was removed and over 90% of the holocellulose was retained. The physicochemical changes after pretreatment were analyzed by confocal laser scanning microscopy, X-ray diffractometer and Fourier transform infrared spectroscopy, showing that the majority of lignin was removed from secondary cell walls and cell middle lamella while the chlorite-resistant lignin remained in the cell corner. Lignin removal significantly enhanced the biodegradability from 59.6% to 86.4% and increased methane production by 38.3%. Energy balance showed that SCA pretreatment was efficient to increase the energy output of hybrid Pennisetum.

In this study, batch experiments were conducted to compare the effect of temperature and substrate source on microbial communities in the anaerobic digestion of kitchen waste. The results showed that the microbial communities of anaerobic digestion were not sensitive to varied sources of waste, but shifted with the change in operating temperatures. In the reactors operated at mesophilic conditions, Levilinea, Syntrophomonas, Methanothrix, and Methanosphaerula, etc. were the dominant microbes during the process. While in thermophilic reactors, Levilinea, Ornatilinea, Methanosphaerula and Methanomassiliicoccus, etc. prevailed. Meanwhile, an enrichment in Coprothermobacter, Defluviitoga, Defluviitalea, Tepidimicrobium, Lutispora and Fonticella were observed as the temperature changed from mesophilic to thermophilic, suggesting these genera could be selectively enriched at thermophilic conditions. The results provided fundamental understanding of the microbiology that could support the scale up of food waste anaerobic digestion.

The effect of additives on the silage quality, microbial community, and anaerobic digestion performance of Pennisetum purpureum with high moisture content was studied. The sample treated with a mixed additive had best silage quality with the lowest pH and highest lactic acid/acetic acid ratio. Different additives influenced the dominant desirable bacteria. Correspondingly, Enterobacter was the dominant bacterial genus for sample with non-fermentative additives, whereas for the samples with fermentative or mixed additives, both Enterobacter and Lactobacillus had high relative abundance. The parameters of NH3-N, hemicellulose and lactic acid were positively correlated with the specific methane yield, while the lignin content was inversely correlated with the specific methane yield. The higher specific methane yield of 293.81 ± 0.15–334.69 ± 22.75 mL/g VS was obtained for samples treated with fermentative additive. Therefore, the mixed additive and fermentative additive are recommended for the silage of material with high-moisture content to improve the silage quality and methane yield.

Material flow analysis (MFA) was applied to study the process of biogas production from switchgrass using a mid-temperature (35 ± 1 °C) batch anaerobic digestion process. The flow distributions of energy and material, including carbon (C) and nitrogen (N), were analyzed, as were the material and energy conversion efficiencies. The results showed that biogas and CH4 production were 268.80 and 135.31 NmLgVS-1 added, respectively, and the average CH4 content in biogas was 50.34%. Based on the MFA of the anaerobic digestion process, 30.6%, 3.6%, and 65.8% of C was converted into biogas, biogas slurry, and biogas residue, respectively; and 11.7% and 88.3% of N was converted into biogas slurry and biogas residue, respectively. The conversion efficiencies of the material and energy from switchgrass to biogas were 36.1% and 30.1%. Because of the low conversion efficiencies of matter and energy during biogas production, it is necessary to strengthen the secondary use of the fermentation residue. This study provides a basis for the optimization of the anaerobic digestion process and efficient utilization of resources and energy of energy-grass.

To investigate whether bioaugmentation could improve the digestion performance of high C/N ratio feedstock without co-digestion with nitrogen-rich substrate, different forms of enriched methanogenic culture were introduced to the continuous feed digesters. The performance efficiency of bioaugmentation on digestion improvement was compared. The effect of bioaugmentation on microbial community composition was revealed as well. Results demonstrated that routine bioaugmentation with liquid culture (containing the microbes and the medium remains) showed the best performance, with the organic loading rate (OLR), methane percentage, volumetric methane production (VMP) and volatile solid methane production (VSMP) higher at 1.0 g L−1 d−1, 24%, 0.22 L L−1 d−1 and 0.23 L g−1 VS d−1 respectively, compared to the non-bioaugmentation control. Whole genome pyrosequencing analysis suggested that consecutive microbial consortium addition could reconstruct the methanogens community by increasing the populations of acetoclastic methanogens Methanothrix, which could accelerate the degradation of acetate and methane production.

Alkaline pretreatment with NaOH was used to improve methane yield from Pennisetum Hybrid. The pretreatments were carried out with different NaOH solutions (2–8% w/w) at three temperatures (35, 55 and 121 °C) for different periods of time (24, 24 and 1 h). All treated and untreated Pennisetum Hybrid were digested under mesophilic conditions (37 °C) to biogas, significant effects of the pretreatments on the yield of methane were observed. Results showed the modified Gompertz equation was reliable (determination coefficients (R2) greater than 0.96) to describe the kinetic behavior of anaerobic digestion of Pennisetum Hybrid. The best result, obtained by the treatment at 35 °C 2% NaOH for 24 h, resulted in the methane yield of 301.7 mL/g VS, corresponding to 21.0% improvement in the methane yield. Compositional, SEM, XRD and FTIR analysis confirmed that lignin removal, structural modification and cellulose crystalline variation were responsible for the improvement.

The silage quality and anaerobic digestion performance of Pennisetum purpereum with molasses-processed wastewater addition were assessed. The silage samples with molasses-processed wastewater addition obtained higher lactic acid concentration of 15.18-23.38mg/g FM, lower pH value of 3.96-4.45 and lower NH3-N content of 0.47-0.64mg/g FM. No obvious difference was observed in the dominant phyla and genus, but the relative abundance of Lactobacillus reached up to 77.39%, increased by 84% compared to the silage samples without molasses-processed wastewater addition. Combined the silage quality and bacterial community, the decreased in pH value of fresh material caused by molasses-processed wastewater addition was the main reason for improving the silage quality. Meanwhile, the increased in COD concentration is beneficial for improving the specific methane yield and the maximum specific methane yield of 259±5.75mL/g VS was obtained with adding 20g/kg molasses-alcoholic wastewater. Molasses-processed wastewater is an alternative additive for silage.

Microbial fuel cell (MFC), which can directly generate electricity from biodegradable materials, has been receiving increasing attention. Effects of temperature change on power density, electrode potential, columbic efficiency, chemical oxygen demand removal and internal resistance in two chambers MFCs were examined in this paper. The maximum power density of 7.89 W/m3 was achieved at 37 degrees C, with 199% higher at 10 degrees C (2.64 W/m3), 24% higher at 30 degrees C (6.34 W/m3) and 21% higher at 43 degrees C, no steady power generation was observed at 55 degrees C. Low temperature to 10 degrees C might have a huge effect on anode potential, especially at higher current, but increasing the temperature to 43 degrees C had a main effect on the cathode performance when the MFCs have been established at 37 degrees C. The internal resistance of MFC was about 29 omega at 37 degrees C, and increased 62% and 303% when MFC switched to 30 degrees C and 10 degrees C. Similarly, internal resistance increased 48% at 43 degrees C. The effect of temperature on MFC performance was expressed by internal resistance, the higher the internal resistance of MFC, the lesser the power density obtained. The Columbic efficiencies were 8.65% at 30 degrees C, 8.53% at 37 degrees C, and 13.24% at 43 degrees C. These results demonstrate that MFCs can effectively be operated over a wide range of temperatures.

Microbial fuel cells (MFCs) are devices that exploit microorganisms as biocatalysts to degrade organic matter or sludge present in wastewater (WW), and thereby generate electricity. We developed a simple, low-cost single-chamber microbial fuel cell (SCMFC)-type biochemical oxygen demand (BOD) sensor using carbon felt (anode) and activated sludge, and demonstrated its feasibility in the construction of a real-time BOD measurement system. Further, the effects of anodic pH and organic concentration on SCMFC performance were examined, and the correlation between BOD concentration and its response time was analyzed. Our results demonstrated that the SCMFC exhibited a stable voltage after 132 min following the addition of synthetic WW (BOD concentration: 200 mg/L). Notably, the response signal increased with an increase in BOD concentration (range: 5–200 mg/L) and was found to be directly proportional to the substrate concentration. However, at higher BOD concentrations (>120 mg/L) the response signal remained unaltered. Furthermore, we optimized the SCMFC using synthetic WW, and tested it with real WW. Upon feeding real WW, the BOD values exhibited a standard deviation from 2.08 to 8.3% when compared to the standard BOD5 method, thus demonstrating the practical applicability of the developed system to real treatment effluents.

The existing energy is almost provided by carbon-containing fossil sources such as oil and coal. The rapid consumption of these fossil resources causes an accelerated release of CO 2 which is a major cause of global warming and associated climate change. “Green” bio-methane (also named biogas) has the advantage for both reducing over dependency on fossil energy and reducing emissions o CO 2 and other greenhouse gases (GHGs). The purpose of this work is to introduce the biogas utilization ways widely applied in China and indicate the development orientation on clean, effective and economical utilizations. Firstly, biogas in china should gradually walk out from rural family, come into centralized anaerobic digestion (CAD). Secondly, combined production of heat and power (CHP) and upgrading to vehicle fuel should be popularized extensively. Thirdly, biogas fuel cell systems and biogas chemical industry (BCI) should be exploited as a technique reservation.