Sandip B. Bankar

Continuous two stage acetone-butanol-ethanol fermentation with integrated solvent removal using Clostridium acetobutylicum B 5313.

The objective of this study was to optimize continuous acetone-butanol-ethanol (ABE) fermentation using a two stage chemostat system integrated with liquid-liquid extraction of solvents produced in the first stage. This minimized end product inhibition by butanol and subsequently enhanced glucose utilization and solvent production in continuous cultures of Clostridium acetobutylicum B 5313. During continuous two-stage ABE fermentation, sugarcane bagasse was used as the cell holding material for the both stages and liquid-liquid extraction was performed using an oleyl alcohol and decanol mixture. An overall solvent production of 25.32g/L (acetone 5.93g/L, butanol 16.90g/L and ethanol 2.48g/L) was observed as compared to 15.98g/L in the single stage chemostat with highest solvent productivity and solvent yield of 2.5g/Lh and of 0.35g/g, respectively. Maximum glucose utilization (83.21%) at a dilution rate of 0.051/h was observed as compared to 54.38% in the single stage chemostat.

Biobutanol: the outlook of an academic and industrialist

The gradual shift of transportation fuels from oil based fuels to alternative fuel resources and worldwide demand for energy has been the impetus for research to produce alcohol biofuels from renewable resources. Currently bioethanol and biodiesel can, however, not cover an increasing demand for biofuels. Hence, there is an extensive need for advanced biofuels with superior fuel properties. The present review is focused on the development of biobutanol, which is regarded to be superior to bioethanol in terms of energy density and hygroscopicity. Although acetone–butanol–ethanol (ABE) fermentation is one of the oldest large-scale fermentation processes, butanol yield by anaerobic fermentation remains sub-optimal. For sustainable industrial scale butanol production, a number of obstacles need to be addressed including choice of feedstock, low product yield, product toxicity to production strain, multiple end-products and downstream processing of alcohol mixtures. Metabolic engineering provides a means for fermentation improvements. Different strategies are employed in the metabolic engineering of Clostridia that aim to enhance the solvent production, improve selectivity for butanol production, and increase the tolerance of Clostridia to solvents. The introduction and expression of a non-clostridial butanol producing pathway in E. coli is a most promising strategy for butanol biosynthesis. Several rigorous kinetic and physiological models for fermentation have been formulated, which form a useful tool for optimization of the process. Due to the lower butanol titers in the fermentation broth, simultaneous fermentation and product removal techniques have been developed to improve production economics. With the use of new strains, inexpensive substrates, and superior reactor designs, economic ABE fermentation may further attract the attention of researchers all over the world. The present review is attempting to provide an overall outlook on discoveries and strategies that are being developed for commercial n-butanol production.

Optimization of poly-ε-lysine production by Streptomyces noursei NRRL 5126

Poly-epsilon-lysine (epsilon-PL) is a non-toxic biopolymer with antimicrobial properties. The production of epsilon-PL by Streptomyces noursei NRRL 5126 shake-flask culture was optimized by identifying the most significant medium components which affect epsilon-PL production (glycerol, proteose peptone and ammonium sulphate) by Placket-Burman design and by application of an evolutionary operation (EVOP) to determine the optimal concentrations of these components. The epsilon-PL yield increased from 41.81 g/l in basal medium to 98.07 g/l in the EVOP-optimized medium containing 3% glycerol, 1% proteose peptone and 0.8% ammonium sulphate. Further improvements in media composition and culture conditions will be required to obtain yields comparable to those obtained with current commercial strains such as Streptomyces albulus.

The two stage immobilized column reactor with an integrated solvent recovery module for enhanced ABE production.

The production of acetone, butanol, and ethanol (ABE) by fermentation is a process that had been used by industries for decades. Two stage immobilized column reactor system integrated with liquid-liquid extraction was used with immobilized Clostridium acetobutylicum DSM 792, to enhance the ABE productivity and yield. The sugar mixture (glucose, mannose, galactose, arabinose, and xylose) representative to the lignocellulose hydrolysates was used as a substrate for continuous ABE production. Maximum total ABE solvent concentration of 20.30 g L(-1) was achieved at a dilution rate (D) of 0.2h(-1), with the sugar mixture as a substrate. The maximum solvent productivity (10.85 g L(-1)h(-1)) and the solvent yield (0.38 g g(-1)) were obtained at a dilution rate of 1.0 h(-1). The maximum sugar mixture utilization rate was achieved with the present set up which is difficult to reach in a single stage chemostat. The system was operated for 48 days without any technical problems.

Enzymatic hydrolysis of hardwood and softwood harvest residue fibers released by sulfur dioxide-ethanol-water fractionation.

The enzymatic hydrolysis of hardwood and softwood harvest residues treated by SO2-ethanol-water (SEW) fractionation was studied. The target was to convert these fibers with high yield into glucose monomers which could be further converted into biofuel by a subsequent fermentation stage. Hardwood biomass residues were efficiently digested at low enzyme dosage (5 FPU/g cellulose) whereas the softwood residues required notably higher enzyme dosage (20 FPU) for sufficient conversion. However, cellulase dosage of softwood could be reduced mannanase supplementation. Especially the high lignin content of softwood biomass pulps impairs the digestibility and thereby, improved delignification could notably enhance the hydrolysis yields. It was shown that inferior delignification of SW biomass is due to persistent polyphenolic acids present in coniferous bark, whereas no evidence of the negative effect of inorganics and acetone extractives was observed. Additionally, SW hydrolyzate was successfully converted into a mixture of butanol, acetone and ethanol through ABE fermentation.

Metabolic precursors enhance the production of poly-ε-lysine by Streptomyces noursei NRRL 5126

Poly‐ε‐lysine produced by streptomyces species is a promising biopolymer owing to its antimicrobial activity and safety for humans. A number of nutritional factors influencing poly‐ε‐lysine production by Streptomyces noursei NRRL 5126 were studied. Various metabolic precursors such as amino acids, tricarboxylic acid cycle intermediates and cofactors were investigated for improved production of poly‐ε‐lysine. Results indicated L‐aspartate (2 mM) and citric acid (5 mM) to substantially increase the poly‐ε‐lysine production from 97.08 to 409.94 mg/L. Addition of citric acid after 24 h and L‐aspartate after 36 h of fermentation medium further enhanced poly‐ε‐lysine production to 497.67 mg/L after a total fermentation time of 108 h. However, the use of cofactors of enzymes involved in the biosynthesis of poly‐ε‐lysine inhibited its production which is believed to be due to diversion of the flux to other metabolites.

Panorama of poly-ε-lysine

Poly-e-lysine (e-PL), (S)-poly(imino(2-amino-1-oxo-1,6-hexanediyl)), is a naturally occurring homopolymer of L-lysine with a degree of polymerization of 25–35, and characterized by peptide bonds between the carboxyl and e-amino groups of L-lysine. e-PL is currently receiving much attention due to its wide applications in chemical, pharmaceutical, food, medical, clinical chemistry, biotechnology and other industries. Moreover, it is water-soluble, biodegradable, heat stable, edible and nontoxic towards humans. The present review discusses the production, recovery, characterization, biodegradation and applications of e-PL. Production of e-PL by fermentation is detailed, along with its industrial problems such as biodegradation. Various purification techniques for higher recovery of e-PL are also described here. Issues of e-PL degrading enzyme and its characterization studies have also been addressed. Molecular genetics as a tool for development of highly productive strains is also discussed herein. Applications of e-PL in various industries showed to have an increasing impact on bioprocessing.

Sustainable biobutanol production from pineapple waste by using Clostridium acetobutylicum B 527: Drying kinetics study

Present investigation explores the use of pineapple peel, a food industry waste, for acetone-butanol-ethanol (ABE) production using Clostridium acetobutylicum B 527. Proximate analysis of pineapple peel shows that it contains 35% cellulose, 19% hemicellulose, and 16% lignin on dry basis. Drying experiments on pineapple peel waste were carried out in the temperature range of 60-120°C and experimental drying data was modeled using moisture diffusion control model to study its effect on ABE production. The production of ABE was further accomplished via acid hydrolysis, detoxification, and fermentation process. Maximum total sugar release obtained by using acid hydrolysis was 97g/L with 95-97% and 10-50% removal of phenolics and acetic acid, respectively during detoxification process. The maximum ABE titer obtained was 5.23g/L with 55.6% substrate consumption when samples dried at 120°C were used as a substrate (after detoxification).

Co-immobilization of glucose oxidase-catalase: Optimization of immobilization parameters to improve the immobilization yield

Co-immobilization of glucose oxidase (EC 1.1.3.4) and catalase (EC 1. 11.1.6) on non-porus glass surfaces using ?-aminopropyltriethoxysilane and polyethyleneimine as an activator and gluteraldehyde as cross linking agent has been carried out for its potential use. Polyethyleneimine was found to be a superior immobilization activator than ?-aminopropyltriethoxysilane. In present study, the effects of rough and smooth beads and optimization of the ratio of enzyme concentrations, activator material and concentration of the cross linking agent were investigated using response surface technology. With optimized concentration of glucose oxidase to catalase ratio (0.97), polyethyleneimine (150 mg/l) and gluteraldehyde (15 ml), the effect of glass bead concentration for maximum immobilization yield was investigated. Central composite optimization strategy with 16 experiments increased immobilization yield from 82.63% to 92.74%. It was observed that 0.3 ml of beads per 120 U of glucose oxidase were necessary for higher immobilization yield.

A green process for the production of butanol from butyraldehyde using alcohol dehydrogenase: process details

Depletion of energy sources has drawn attention towards production of bio-butanol by fermentation. However, the process is constrained by product inhibition which results in low product yield. Hence, a new strategy wherein butanol was produced from butyraldehyde using alcohol dehydrogenase and NADH as a cofactor was developed. Butyraldehyde can be synthesized chemically or through fermentation. The problem of cofactor regeneration during the reaction for butanol production was solved using substrate coupled and enzyme coupled reactions. The conventional reaction produced 35% of butanol without regeneration of cofactor using 300 μM NADH. The process of substrate coupled reaction was optimized to get maximum conversion. NADH (30 μM) and 100 μg per ml of alcohol dehydrogenase (320 U mg−1) could convert 17.39 mM of butyraldehyde to butanol using ethanol (ratio of butyraldehye to ethanol 1 : 4) giving a maximum conversion of 75%. The enzyme coupled reaction under the same conditions showed only 24% conversion of butyraldehyde to butanol using the glutamate dehydrogenase-L-glutamate enzyme system for the regeneration of cofactor. Hence, substrate coupled reaction is suggested as a better method over the enzyme coupled reaction for the cost effective production of butanol.