Feng-Wu Bai

Prospective and development of butanol as an advanced biofuel.

Butanol has been acknowledged as an advanced biofuel, but its production through acetone-butanol-ethanol (ABE) fermentation by clostridia is still not economically competitive, due to low butanol yield and titer. In this article, update progress in butanol production is reviewed. Low price and sustainable feedstocks such as lignocellulosic residues and dedicated energy crops are needed for butanol production at large scale to save feedstock cost, but processes are more complicated, compared to those established for ABE fermentation from sugar- and starch-based feedstocks. While rational designs targeting individual genes, enzymes or pathways are effective for improving butanol yield, global and systems strategies are more reasonable for engineering strains with stress tolerance controlled by multigenes. Compared to solvent-producing clostridia, engineering heterologous species such as Escherichia coli and Saccharomyces cerevisiae with butanol pathway might be a solution for eliminating the formation of major byproducts acetone and ethanol so that butanol yield can be improved significantly. Although batch fermentation has been practiced for butanol production in industry, continuous operation is more productive for large scale production of butanol as a biofuel, but a single chemostat bioreactor cannot achieve this goal for the biphasic ABE fermentation, and tanks-in-series systems should be optimized for alternative feedstocks and new strains. Moreover, energy saving is limited for the distillation system, even total solvents in the fermentation broth are increased significantly, since solvents are distilled to ~40% by the beer stripper, and more than 95% water is removed with the stillage without phase change, even with conventional distillation systems, needless to say that advanced chemical engineering technologies can distil solvents up to ~90% with the beer stripper, and the multistage pressure columns can well balance energy consumption for solvent fraction. Indeed, an increase in butanol titer with ABE fermentation can significantly save energy consumption for medium sterilization and stillage treatment, since concentrated medium can be used, and consequently total mass flow with production systems can be reduced. As for various in situ butanol removal technologies, their energy efficiency, capital investment and contamination risk to the fermentation process need to be evaluated carefully.

High-titer n-butanol production by clostridium acetobutylicum JB200 in fed-batch fermentation with intermittent gas stripping

Acetone–butanol–ethanol (ABE) fermentation with a hyper‐butanol producing Clostridium acetobutylicum JB200 was studied for its potential to produce a high titer of butanol that can be readily recovered with gas stripping. In batch fermentation without gas stripping, a final butanol concentration of 19.1 g/L was produced from 86.4 g/L glucose consumed in 78 h, and butanol productivity and yield were 0.24 g/L h and 0.21 g/g, respectively. In contrast, when gas stripping was applied intermittently in fed‐batch fermentation, 172 g/L ABE (113.3 g/L butanol, 49.2 g/L acetone, 9.7 g/L ethanol) were produced from 474.9 g/L glucose in six feeding cycles over 326 h. The overall productivity and yield were 0.53 g/L h and 0.36 g/g for ABE and 0.35 g/L h and 0.24 g/g for butanol, respectively. The higher productivity was attributed to the reduced butanol concentration in the fermentation broth by gas stripping that alleviated butanol inhibition, whereas the increased butanol yield could be attributed to the reduced acids accumulation as most acids produced in acidogenesis were reassimilated by cells for ABE production. The intermittent gas stripping produced a highly concentrated condensate containing 195.9 g/L ABE or 150.5 g/L butanol that far exceeded butanol solubility in water. After liquid–liquid demixing or phase separation, a final product containing ∼610 g/L butanol, ∼40 g/L acetone, ∼10 g/L ethanol, and no acids was obtained. Compared to conventional ABE fermentation, the fed‐batch fermentation with intermittent gas stripping has the potential to reduce at least 90% of energy consumption and water usage in n‐butanol production from glucose. Biotechnol. Bioeng. 2012; 109: 2746–2756.

Two-stage in situ gas stripping for enhanced butanol fermentation and energy-saving product recovery.

Two-stage gas stripping for butanol recovery from acetone-butanol-ethanol (ABE) fermentation with Clostridium acetobutylicum JB200 in a fibrous bed bioreactor was studied. Compared to fermentation without in situ gas stripping, more ABE (10.0 g/L acetone, 19.2 g/L butanol, 1.7 g/L ethanol vs. 7.9 g/L acetone, 16.2 g/L butanol, 1.4 g/L ethanol) were produced, with a higher butanol yield (0.25 g/g vs. 0.20 g/g) and productivity (0.40 g/L·h vs. 0.30 g/L·h) due to reduced butanol inhibition. The first-stage gas stripping produced a condensate containing 175.6 g/L butanol (227.0 g/L ABE), which after phase separation formed an organic phase containing 612.3g/L butanol (660.7 g/L ABE) and an aqueous phase containing 101.3 g/L butanol (153.2 g/L ABE). After second-stage gas stripping, a highly concentrated product containing 420.3 g/L butanol (532.3 g/L ABE) was obtained. The process is thus effective in producing high-titer butanol that can be purified with much less energy.

Yeast flocculation: New story in fuel ethanol production

Yeast flocculation has been used in the brewing industry to facilitate biomass recovery for a long time, and thus its mechanism of yeast flocculation has been intensively studied. However, the application of flocculating yeast in ethanol production garnered attention mainly in the 1980s and 1990s. In this article, updated research progress in the molecular mechanism of yeast flocculation and the impact of environmental conditions on yeast flocculation are reviewed. Construction of flocculating yeast strains by genetic approach and utilization of yeast flocculation for ethanol production from various feedstocks were presented. The concept of self-immobilized yeast cells through their flocculation is revisited through a case study of continuous ethanol fermentation with the flocculating yeast SPSC01, and their technical and economic advantages are highlighted by comparing with yeast cells immobilized with supporting materials and regular free yeast cells as well. Taking the flocculating yeast SPSC01 as an example, the ethanol tolerance of the flocculating yeast was also discussed.

Current progress and future prospect of microalgal biomass harvest using various flocculation technologies

Microalgae have been extensively studied for the production of various valuable products. Application of microalgae for the production of renewable energy has also received increasing attention in recent years. However, high cost of microalgal biomass harvesting is one of the bottlenecks for commercialization of microalgae-based industrial processes. Considering harvesting efficiency, operation economics and technological feasibility, flocculation is a superior method to harvest microalgae from mass culture. In this article, the latest progress of various microalgal cell harvesting methods via flocculation is reviewed with the emphasis on the current progress and prospect in environmentally friendly bio-based flocculation. Harvesting microalgae through bio-based flocculation is a promising component of the low-cost microalgal biomass production technology.

Bioflocculant production from Solibacillus silvestris W01 and its application in cost-effective harvest of marine microalga Nannochloropsis oceanica by flocculation

Microalgae are widely studied for biofuel production, however, current technologies to harvest microalgae for this purpose are not well developed. In this work, a bacterial strain W01 was isolated from activated sludge and identified as Solibacillus silvestris. Bioflocculant in the culture broth of W01 showed 90% flocculating efficiency on marine microalga Nannochloropsis oceanica, and no metal ion was required for the flocculation process. Chemical analysis of the purified bioflocculant indicated that it is a proteoglycan composed of 75.1% carbohydrate and 24.9% protein (w/w). The bioflocculant exhibits no effect on the growth of microalgal cells and can be reused to for economical harvesting of N. oceanica. This is the first report that strain of S. silvestris can produce bioflocculant for microalgae harvest. The novel bioflocculant produced by W01 has the potential to harvest marine microalgae for cost-effective production of microalgal bioproducts.

Redox potential control and applications in microaerobic and anaerobic fermentations

Many fermentation products are produced under microaerobic or anaerobic conditions, in which oxygen is undetectable by dissolved oxygen probe, presenting a challenge for process monitoring and control. Extracellular redox potentials that can be detected conveniently affect intracellular redox homeostasis and metabolism, and consequently control profiles of fermentation products, which provide an alternative for monitoring and control of these fermentation processes. This article reviews updated progress in the impact of redox potentials on gene expression, protein biosynthesis and metabolism as well as redox potential control strategies for more efficient production of fermentation products, taking ethanol fermentation by the yeast Saccharomyces under microaerobic conditions and butanol production by the bacterium Clostridium under anaerobic conditions as examples.

A novel in situ gas stripping‐pervaporation process integrated with acetone‐butanol‐ethanol fermentation for hyper n‐butanol production

Butanol is considered as an advanced biofuel, the development of which is restricted by the intensive energy consumption of product recovery. A novel two‐stage gas stripping‐pervaporation process integrated with acetone‐butanol‐ethanol (ABE) fermentation was developed for butanol recovery, with gas stripping as the first‐stage and pervaporation as the second‐stage using the carbon nanotubes (CNTs) filled polydimethylsiloxane (PDMS) mixed matrix membrane (MMM). Compared to batch fermentation without butanol recovery, more ABE (27.5 g/L acetone, 75.5 g/L butanol, 7.0 g/L ethanol vs. 7.9 g/L acetone, 16.2 g/L butanol, 1.4 g/L ethanol) were produced in the fed‐batch fermentation, with a higher butanol productivity (0.34 g/L · h vs. 0.30 g/L · h) due to reduced butanol inhibition by butanol recovery. The first‐stage gas stripping produced a condensate containing 155.6 g/L butanol (199.9 g/L ABE), which after phase separation formed an organic phase containing 610.8 g/L butanol (656.1 g/L ABE) and an aqueous phase containing 85.6 g/L butanol (129.7 g/L ABE). Fed with the aqueous phase of the condensate from first‐stage gas stripping, the second‐stage pervaporation using the CNTs‐PDMS MMM produced a condensate containing 441.7 g/L butanol (593.2 g/L ABE), which after mixing with the organic phase from gas stripping gave a highly concentrated product containing 521.3 g/L butanol (622.9 g/L ABE). The outstanding performance of CNTs‐PDMS MMM can be attributed to the hydrophobic CNTs giving an alternative route for mass transport through the inner tubes or along the smooth surface of CNTs. This gas stripping‐pervaporation process with less contaminated risk is thus effective in increasing butanol production and reducing energy consumption. Biotechnol. Bioeng. 2016;113: 120–129.

Impact of zinc supplementation on the improvement of ethanol tolerance and yield of self-flocculating yeast in continuous ethanol fermentation

The effects of zinc supplementation were investigated in the continuous ethanol fermentation using self-flocculating yeast. Zinc sulfate was added at the concentrations of 0.01, 0.05 and 0.1 g l(-1), respectively. Reduced average floc sizes were observed in all the zinc-supplemented cultures. Both the ethanol tolerance and thermal tolerance were significantly improved by zinc supplements, which correlated well with the increased ergosterol and trehalose contents in the yeast flocs. The highest ethanol concentration by 0.05 g l(-1) zinc sulfate supplementation attained 114.5 g l(-1), in contrast to 104.1 g l(-1) in the control culture. Glycerol production was decreased by zinc supplementations, with the lowest level 3.21 g l(-1), about 58% of the control. Zinc content in yeast cells was about 1.4 microMol g(-1) dry cell weight, about sixfold higher than that of control in all the zinc-supplemented cultures, and close correlation of zinc content in yeast cells with the cell viability against ethanol and heat shock treatment was observed. These studies suggest that exogenous zinc addition led to a reprogramming of cellular metabolic network, resulting in enhanced ethanol tolerance and ethanol production.

Bioethanol from Lignocellulosic Biomass

China is suffering from a sustained shortage of crude oil supply, making fuel ethanol and other biofuels alternative solutions for this issue. However, taking into account the countrys large population and dwindling arable land due to rapid urbanization, it is apparent that current fuel ethanol production from grain-based feedstocks is not sustainable, and lignocellulosic biomass, particularly agricultural residues that are abundantly available in China, is the only choice for China to further expand its fuel ethanol production, provided economically viable processes can be developed. In this chapter, cutting edge progress in bioethanol is reviewed, with a focus on the understanding of the molecular structure of the feedstock, leading pretreatment technologies, enzymatic hydrolysis of the cellulose component and strategies for the co-fermentation of the C5 and C6 sugars with engineered microorganisms. Finally, process integration and optimization is addressed with a case study on the COFCO Corporations pilot plant, and challenges and perspectives for commercial production of bioethanol are highlighted.