Lijie Chen

Butanol derived from renewable resources by microbial fermentation is considered as one of not only valuable platform chemicals but alternative advanced biofuels. However, due to low butanol concentration in fermentation broth, butanol production is restricted by high energy consumption for product recovery. For in situ butanol recovery techniques, such as gas stripping and pervaporation, the common problem is their low efficiency in harvesting and concentrating butanol. Therefore, there is a necessity to develop an advanced butanol recovery technique for cost-effective biobutanol production. A close-circulating vapor stripping-vapor permeation (VSVP) process was developed with temperature-difference control for single-stage butanol recovery. In the best scenario, the highest butanol separation factor of 142.7 reported to date could be achieved with commonly used polydimethylsiloxane membrane, when temperatures of feed solution and membrane surroundings were 70 and 0 °C, respectively. Additionally, more ABE (31.2 vs. 17.7 g/L) were produced in the integrated VSVP process, with a higher butanol yield (0.21 vs. 0.17 g/g) due to the mitigation of butanol inhibition. The integrated VSVP process generated a highly concentrated permeate containing 212.7 g/L butanol (339.3 g/L ABE), with the reduced energy consumption of 19.6 kJ/g-butanol. Therefore, the present study demonstrated a well-designed energy-efficient technique named by vapor stripping-vapor permeation for single-stage butanol removal. The butanol separation factor was multiplied by the temperature-difference control strategy which could double butanol recovery performance. This advanced VSVP process can completely eliminate membrane fouling risk for fermentative butanol separation, which is superior to other techniques.

Butanol as an important chemical and potential fuel could be produced via ABE fermentation from lignocellulosic biomass. The use of food-related feedstocks such as maize and sugar cane may not be a sustainable solution to world’s energy needs. Recently, Jerusalem artichoke tubers containing inulin have been used as feedstock for butanol production, but this bioprocess is not commercially feasible due to the great value of inulin as functional food. Till now, there is a gap on the utilization of Jerusalem artichoke stalk (JAS) as feedstock for microbial butanol production. Biobutanol production from JAS was investigated in order to improve cellulose digestibility and efficient biobutanol fermentation. Compared with 9.0 g/L butanol (14.7 g/L ABE) production by 2% NaOH pretreatment of JAS, 11.8 g/L butanol (17.6 g/L ABE) was produced in the best scenario conditions of NaOH–H2O2 pretreatment, washing times and citrate buffer strengths etc. Furthermore, more than >64% water in washing pretreated JAS process could be saved, with improving butanol production by >25.0%. To mimic in situ product recovery for ABE fermentation, the vapor stripping–vapor permeation (VSVP) process steadily produced 323.4–348.7 g/L butanol (542.7–594.0 g/L ABE) in condensate, which showed more potentials than pervaporation for butanol recovery. Therefore, the present study demonstrated an effective strategy on efficient biobutanol production using lignocellulosic biomass. The process optimization could contribute to significant reduction of wastewater emission and the improvement of lignocellulosic biomass digestibility and biobutanol production, which makes biobutanol production more efficient using JAS.

Catalytic conversion of lignocellulosic biomass to high-value transportation petrol, jet and diesel fuels is of great importance to develop versatile renewable energy and boost the rural economy, thus it is receiving worldwide attention. However, the state-of-the-art processes still suffer from a low efficiency in yield and productivity, which hinders the progress of its commercialization. Thus, it is essential to explore the consolidated catalytic reaction and refinery process. Here we report an advanced approach/process that could continuously produce long-chain (C₅–C₁₅) ketones from lignocellulosic biomass in a cascade mode by integrating several bio- and chemical catalysis reactions/processes, which exhibits a high efficiency and stable conversion rate. This technology consolidated the acetone–butanol–ethanol (ABE) fermentation, palladium catalyzed alkylation and in situ product recovery, allowing the catalytic reaction to proceed consistently and smoothly. The remarkable conversion properties are mainly attributed to the excellent compatible linkage of chemical catalysis with biosynthesis and process design to stabilize the Pd catalytic reaction. This validated strategy for the rational process design and integration of chemical- and bio-catalysis offers great demonstration in producing high-value transportation biofuels derived from agricultural residues.

Butanol is not only an important solvent and chemical intermediate in food and pharmaceutical industries, but also considered as an advanced biofuel. Recently, there have been resurging interests in producing biobutanol especially using low-cost lignocellulosic biomass, but the process still suffers from low titer and productivity. The challenge for the bioconversion approach is to find an effective way of degrading materials into simple sugars that can then be converted into fuels by microorganisms. The pretreatment of lignocellulosic biomass is the great important process in influencing butanol production and recovery, finally determining its eco-feasibility in commercialization. The effects of various strengths of citrate buffer on enzymatic hydrolysis and acetone–butanol–ethanol fermentation using corn stover or glucose as feedstock were investigated. The strengths of citrate buffer in the range of 20–100 mM had no effect on enzymatic hydrolysis, but greatly influenced the performance of ABE fermentation using corn stover hydrolysate. When 30 mM citrate buffer was used for enzymatic hydrolysis, the fermentation broth with the maximum butanol and ABE concentrations of 11.2 and 19.8 g/L were obtained from 30.9 g/L glucose and 9.7 g/L xylose, respectively, which was concentrated to 100.4 g/L butanol and 153.5 g/L ABE by vapor stripping–vapor permeation process. Furthermore, using glucose as sole carbon source, there were no cell growth and ABE production in the P2 medium with 80 or 100 mM citrate buffer, indicating that higher concentrations of citrate buffer had deleterious effect on cell growth and metabolism due to the variation of cells internal pH and cell membrane permeability. To mimic in situ product recovery for ABE fermentation, the VSVP process produced the condensate containing 212.0–232.0 g/L butanol (306.6–356.1 g/L ABE) from fermentation broth containing ~10 g/L butanol (~17 g/L ABE), the performance of which was more effective than pervaporation and gas stripping. As it has significant impact on butanol fermentation, the strength of citrate buffer is of great importance in lignocellulosic butanol fermentation. Compared with pervaporation and gas stripping, the VSVP process has great potential for efficient butanol recovery in biobutanol production.

Biobutanol outperforms bioethanol as an advanced biofuel, but is not economically competitive in terms of its titer, yield and productivity associated with feedstocks and energy cost. In this work, the synergistic effect of calcium and zinc was investigated in the acetone-butanol-ethanol (ABE) fermentation by Clostridium acetobutylicum using glucose, xylose and glucose/xylose mixtures as carbon source(s). Significant improvements associated with enhanced glucose/xylose utilization, cell growth, acids re-assimilation and butanol biosynthesis were achieved. Especially, the maximum butanol and ABE production of 16.1 and 25.9 g L⁻¹ were achieved from 69.3 g L⁻¹ glucose with butanol/ABE productivities of 0.40 and 0.65 g L⁻¹ h⁻¹ compared to those of 11.7 and 19.4 g/L with 0.18 and 0.30 g L⁻¹ h⁻¹ obtained in the control respectively without any supplement. More importantly, zinc was significantly involved in the butanol tolerance based on the improved xylose utilization under various butanol-shock conditions (2, 4, 6, 8 and 10 g L⁻¹ butanol). Under the same conditions, calcium and zinc co-supplementation led to the best xylose utilization and butanol production. These results suggested that calcium and zinc could play synergistic roles improving ABE fermentation by C. acetobutylicum.

The micronutrient zinc plays vital roles in ABE fermentation by Clostridium acetobutylicum. In order to elucidate the zinc-associated response for enhanced glucose utilization and earlier solventogenesis, transcriptional analysis was performed on cells grown in glucose medium at the exponential growth phase of 16 h without/with supplementary zinc. Correspondingly, the gene glcG (CAC0570) encoding a glucose-specific PTS was significantly upregulated accompanied with the other two genes CAC1353 and CAC1354 for glucose transport in the presence of zinc. Additionally, genes involved in the metabolisms of six other carbohydrates (maltose, cellobiose, fructose, mannose, xylose and arabinose) were differentially expressed, indicating that the regulatory effect of micronutrient zinc is carbohydrate-specific with respects to the improved/inhibited carbohydrate utilization. More importantly, multiple genes responsible for glycolysis (glcK and pykA), acidogenesis (thlA, crt, etfA, etfB and bcd) and solventogenesis (ctfB and bdhA) of C. acetobutylicum prominently responded to the supplementary zinc at differential expression levels. Comparative analysis of intracellular metabolites revealed that the branch node intermediates such as acetyl-CoA, acetoacetyl-CoA, butyl-CoA, and reducing power NADH remained relatively lower whereas more ATP was generated due to enhanced glycolysis pathway and earlier initiation of solventogenesis, suggesting that the micronutrient zinc-associated response for the selected intracellular metabolisms is significantly pleiotropic.

Butanol is regarded as an advanced biofuel that can be derived from renewable biomass. However, the main challenge for microbial butanol production is low butanol titer, yield and productivity, leading to intensive energy consumption in product recovery. Various alternative separation technologies such as extraction, adsorption and gas stripping, etc., could be integrated with acetone–butanol–ethanol (ABE) fermentation with improving butanol productivity, but their butanol selectivities are not satisfactory. The membrane-based pervaporation technology is recently attracting increasing attention since it has potentially desirable butanol selectivity. The performance of the zeolite-mixed polydimethylsiloxane (PDMS) membranes were evaluated to recover butanol from butanol/water binary solution as well as fermentation broth in the integrated ABE fermentation system. The separation factor and butanol titer in permeate of the zeolite-mixed PDMS membrane were up to 33.0 and 334.6 g/L at 80°C, respectively, which increased with increasing zeolite loading weight in the membrane as well as feed temperature. The enhanced butanol separation factor was attributed to the hydrophobic zeolites with large pore size providing selective routes preferable for butanol permeation. In fed-batch fermentation incorporated with pervaporation, 54.9 g/L ABE (34.5 g/L butanol, 17.0 g/L acetone and 3.4 g/L ethanol) were produced from 172.3 g/L glucose. The overall butanol productivity and yield increased by 16.0 and 11.1%, respectively, which was attributed to the alleviated butanol inhibition by pervaporation and reassimilation of acids for ABE production. The zeolite-mixed membrane produced a highly concentrated condensate containing 169.6 g/L butanol or 253.3 g/L ABE, which after phase separation easily gave the final product containing >600 g/L butanol. Zeolite loading in the PDMS matrix was attributed to improving the pervaporative performance of the membrane, showing great potential to recover butanol with high purity. Therefore, this zeolite-mixed PDMS membrane had the potential to improve biobutanol production when integrating with ABE fermentation.