Jinglan Wu

Enhanced butanol production by modulation of electron flow in Clostridium acetobutylicum B3 immobilized by surface adsorption

The objective of this study was to improve butanol yield and productivity by redox modulation and immobilization of Clostridium acetobutylicum B3 cells. Stoichiometric network analysis revealed that NAD(P)H that had escaped from the fermentation as H2 limited the butanol yield and led to the accumulation of oxidation byproducts, e.g., acetone. Methyl viologen was used as an electron carrier to divert the electron flow away from H2 production and to reinforce the NAD(P)H supply. Butanol yield was increased by 37.8% with severely diminished acetone production. Immobilization of the cells by adsorption onto a fibrous matrix improved their butanol tolerance and production rate. An average of 15.6 g/L butanol was achieved within 12 h with a solvent productivity of 1.88 g/L/h in repeated batch fermentation. To our knowledge, this is the highest solvent productivity with a relatively high butanol titer produced by a Clostridium strain in batch fermentation.

Selective separation of biobutanol from acetone-butanol-ethanol fermentation broth by means of sorption methodology based on a novel macroporous resin.

The traditional distillation method for recovery of butanol from fermentation broth is an energy‐intensive process. Separation of butanol based on adsorption methodology has advantages in terms of biocompatibility and stability, as well as economy, and therefore gains much attention. However, the application of the commercial adsorbents in the integrated acetone–butanol–ethanol (ABE) fermentation process is restricted due to the low recovery (less than 85%) and the weak capability of enrichment in the eluent (3–4 times). In this study, we investigated the sorption properties of butanol onto three kinds of adsorbents with different polarities developed in our laboratory, that is, XD‐41, H‐511, and KA‐I resin. The sorption behaviors of single component and ABE ternary mixtures presented in the fermentation broths on KA‐I resin were investigated. KA‐I resin had higher affinity for butanol than for acetone, ethanol, glucose, acetic acid, and butyric acid. Multicomponent ABE sorption on KA‐I resin was modeled using a single site extended Langmuir isotherm model. In a desorption study, all the adsorbed components were desorbed in one bed volume of methanol, and the recovery of butanol from KA‐I resin was 99.7%. The concentration of butanol in the eluent was increased by a factor of 6.13. In addition, KA‐I resin was successfully regenerated by two bed volumes of water. Because of its quick sorption, high sorption capacity, low cost, and ease of desorption and regeneration, KA‐I resin exhibits good potential for compatibility with future ABE fermentation coupled with in situ recovery product removal techniques.

Enhancement of n-butanol production by in situ butanol removal using permeating-heating-gas stripping in acetone-butanol-ethanol fermentation.

Butanol recovery from acetone-butanol-ethanol (ABE) fed-batch fermentation using permeating-heating-gas was determined in this study. Fermentation was performed with Clostridium acetobutylicum B3 in a fibrous bed bioreactor and permeating-heating-gas stripping was used to eliminate substrate and product inhibition, which normally restrict ABE production and sugar utilization to below 20 g/L and 60 g/L, respectively. In batch fermentation (without permeating-heating-gas stripping), C. acetobutylicum B3 utilized 60 g/L glucose and produced 19.9 g/L ABE and 12 g/L butanol, while in the integrated process 290 g/L glucose was utilized and 106.27 g/L ABE and 66.09 g/L butanol were produced. The intermittent gas stripping process generated a highly concentrated condensate containing approximately 15% (w/v) butanol, 4% (w/v) acetone, a small amount of ethanol (<1%), and almost no acids, resulting in a highly concentrated butanol solution [∼ 70% (w/v)] after phase separation. Butanol removal by permeating-heating-gas stripping has potential for commercial ABE production.

Experimental and Modeling Studies on the Sorption Breakthrough Behaviors of Butanol from Aqueous Solution in a Fixed-bed of KA-I Resin

Removal of biobutanol from acetone-butanolethanol (ABE) fermentation broth can be achieved by fixed-bed sorption by means of KA-I resin, and the relevant breakthrough curves would provide much valuable information to help design a continuous fixed-bed sorption process in field application. In the present study, the effects of several important design parameters, i.e., initial butanol concentration (Cf: 3.0 ∼ 30.0 g/L), inlet flow rate (Qf: 0.5 ∼ 5.5 mL/min) and adsorbent bed height (Z: 4.2 ∼ 18.0 cm), on the adsorption breakthrough curves of KA-I resin in a fixed-bed column were investigated. It was found that the amount of adsorbed butanol at breakthrough point was increased with an increase in the value of Cf and Z; and with decrease in the value of Qf. However, the maximum sorption capacities of butanol at saturated point were basically unchanged. Three well-established fixed-bed adsorption models, namely Thomas, Yoon-Nelson and Adams-Bohart, were applied to predict the breakthrough curves and to determine the characteristic parameters of fixed-bed column, which are the basis for the process design at a real scale. Good agreement between the theoretical breakthrough curves and the experimental result were observed using Thomas and Yoon-Nelson models.

Modeling of breakthrough curves of single and quaternary mixtures of ethanol, glucose, glycerol and acetic acid adsorption onto a microporous hyper-cross-linked resin

The adsorption of quaternary mixtures of ethanol/glycerol/glucose/acetic acid onto a microporous hyper-cross-linked resin HD-01 was studied in fixed beds. A mass transport model based on film solid linear driving force and the competitive Langmuir isotherm equation for the equilibrium relationship was used to develop theoretical fixed bed breakthrough curves. It was observed that the outlet concentration of glucose and glycerol exceeded the inlet concentration (c/c0>1), which is an evidence of competitive adsorption. This phenomenon can be explained by the displacement of glucose and glycerol by ethanol molecules, owing to more intensive interactions with the resin surface. The model proposed was validated using experimental data and can be capable of foresee reasonably the breakthrough curve of specific component under different operating conditions. The results show that HD-01 is a promising adsorbent for recovery of ethanol from the fermentation broth due to its large capacity, high selectivity, and rapid adsorption rate.

Metallo-deuteroporphyrin as a biomimetic catalyst for the catalytic oxidation of lignin to aromatics.

A series of metallo-deuteroporphyrins derived from hemin were prepared as models of the cytochrome P450 enzyme. With the aid of the highly active Co(II) deuteroporphyrin complex, the catalytic oxidation system was applied for the oxidation of several lignin model compounds, and high yields of monomeric products were obtained under mild reaction conditions. It was found that the modified cobalt deuteroporphyrin that has no substituents at the meso sites but does have the disulfide linkage in the propionate side chains at the β sites exhibited much higher activity and stability than the synthetic tetraphenylporphyrin. The changes in the propionate side chains can divert the reactivity of cobalt deuteroporphyrins from the typical CC bond cleavage to CO bond cleavage. Furthermore, this novel oxidative system can convert enzymolysis lignin into depolymerized products including a significant portion of well-defined aromatic monomers.

A mild and highly efficient laccase-mediator system for aerobic oxidation of alcohols

With the aid of the highly active nitroxyl radical AZADO (2-azaadamantane N-oxyl), a simple method for the aerobic catalytic oxidation of alcohols is presented. The oxidations could typically proceed under practical ambient conditions (room temperature, air atmosphere, no moisture effect, metal-free, etc.) with a broad generality of the alcohol substrates, and especially for the oxidation of complex and highly functionalized alcohols. An ionic mechanism is proposed for the present system.

Production of liquid hydrocarbon fuels with acetoin and platform molecules derived from lignocellulose

Acetoin, a novel C4 platform molecule derived from new ABE (acetoin–butanol–ethanol) type fermentation via metabolic engineering, was used for the first time as a bio-based building block for the production of liquid hydrocarbon fuels. A series of diesel or jet fuel range C9–C14 straight, branched, or cyclic alkanes were produced in excellent yields by means of C–C coupling followed by hydrodeoxygenation reactions. Hydroxyalkylation/alkylation of acetoin with 2-methylfuran was investigated over a series of solid acid catalysts. Among the investigated candidates, zirconia supported trifluoromethanesulfonic acid showed the highest activity and stability. In the aldol condensation step, a basic ionic liquid [H3N+–CH2–CH2–OH][CH3COO−] was identified as an efficient and recyclable catalyst for the reactions of acetoin with furan based aldehydes. The scope of the process has also been studied by reacting acetoin with other aldehydes, and it was found that abnormal condensation products were formed from the reactions of acetoin with aromatic aldehydes through an aldol condensation–pinacol rearrangement route when amorphous aluminium phosphate was used as a catalyst. And the final hydrodeoxygenation step could be achieved by using a simple and handy Pd/C + H-beta zeolite system, and no or a negligible amount of oxygenates was observed after the reaction. Excellent selectivity was also observed using the present system, and the clean formation of hydrocarbons with a narrow distribution of alkanes occurred in most cases.

Acetone-butanol-ethanol competitive sorption simulation from single, binary, and ternary systems in a fixed-bed of KA-I resin.

Separation of butanol based on sorption methodology from acetone–butanol–ethanol (ABE) fermentation broth has advantages in terms of biocompatibility and stability, as well as economy, and therefore gains much attention. In this work a chromatographic column model based on the solid film linear driving force approach and the competitive Langmuir isotherm equations was used to predict the competitive sorption behaviors of ABE single, binary, and ternary mixture. It was observed that the outlet concentration of weaker retained components exceeded the inlet concentration, which is an evidence of competitive adsorption. Butanol, the strongest retained component, could replace ethanol almost completely and also most of acetone. In the end of this work, the proposed model was validated by comparison of the experimental and predicted ABE ternary breakthrough curves using the real ABE fermentation broth as a feed solution.

Construction and expression of a polycistronic plasmid encoding N-acetylglucosamine 2-epimerase and N-acetylneuraminic acid lyase simultaneously for production of N-acetylneuraminic acid.

Synthesis of N-acetylneuraminic acid (Neu5Ac) from N-acetylglucosamine (GlcNAc) and pyruvate was carried out by constructing and expressing a polycistronic plasmid encoding an N-acetylglucosamine 2-epimerase (AGE) gene and an N-acetylneuraminic acid lyase (Nal) gene simultaneously. Nal from Escherichia coli K12 and AGEs from Synechocystis sp. PCC 6803 (snAGE) and Anabaena sp. CH1 (anAGE) were used. And four polycistronic plasmids were constructed in which the positions of AGE gene differed with respect to Nal gene. Among these plasmids, pET-28a-Nal-anAGE with anAGE gene located next to Nal gene caused the production of the highest amount of Neu5Ac, generating 61.3g/L in 60h by whole-cell catalysis without the addition of ATP as AGE activator. And pET-28a-Nal-anAGE lowered anAGEs expression level, allowing it to fold properly. Thus, an inclusion-body-free E. coli strain capable of producing Neu5Ac by whole-cell catalysis with high yield and low cost was constructed in the present study.