Shrikant A. Survase

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.

Butanol production from lignocellulosics

Clostridium spp. produce n-butanol in the acetone/butanol/ethanol process. For sustainable industrial scale butanol production, a number of obstacles need to be addressed including choice of feedstock, the low product yield, toxicity to production strain, multiple-end products and downstream processing of alcohol mixtures. This review describes the use of lignocellulosic feedstocks, bioprocess and metabolic engineering, downstream processing and catalytic refining of n-butanol.

Clavulanic acid: A review

Natural antibiotics are almost universal secondary metabolites, not essential for the growth of the producing organisms generally produced at low growth rates or after growth has ceased. Clavulanic acid (CA), a naturally occurring powerful inhibitor of bacterial beta-lactamases is a major beta-lactam antibiotic produced by organism Streptomyces clavuligerus and is active against a wide spectrum of Gram-positive and Gram-negative bacteria. The review discusses the biosynthetic pathway, fermentative production, downstream processing and applications of CA.

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.

Cyclosporin A — A review on fermentative production, downstream processing and pharmacological applications

In present times, the immunosuppressants have gained considerable importance in the world market. Cyclosporin A (CyA) is a cyclic undecapeptide with a variety of biological activities including immunosuppressive, anti-inflammatory, antifungal and antiparasitic properties. CyA is produced by various types of fermentation techniques using Tolypocladium inflatum. In the present review, we discuss the biosynthetic pathway, fermentative production, downstream processing and pharmacological activities of CyA.

Continuous acetone-butanol-ethanol fermentation using SO2-ethanol-water spent liquor from spruce

SO2-ethanol-water (SEW) spent liquor from spruce chips was successfully used for batch and continuous production of acetone, butanol and ethanol (ABE). Initially, batch experiments were performed using spent liquor to check the suitability for production of ABE. Maximum concentration of total ABE was found to be 8.79 g/l using 4-fold diluted SEW liquor supplemented with 35 g/l of glucose. The effect of dilution rate on solvent production, productivity and yield was studied in column reactor consisting of immobilized Clostridium acetobutylicum DSM 792 on wood pulp. Total solvent concentration of 12 g/l was obtained at a dilution rate of 0.21 h(-1). The maximum solvent productivity (4.86 g/l h) with yield of 0.27 g/g was obtained at dilution rate of 0.64 h(-1). Further, to increase the solvent yield, the unutilized sugars were subjected to batch fermentation.

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.

Oil palm empty fruit bunch to biofuels and chemicals via SO2–ethanol–water fractionation and ABE fermentation

A process has been developed for conversion of spent liquor produced by SO2-ethanol-water (SEW) fractionation of oil palm empty fruit bunch (OPEFB) fibers to biofuels by ABE fermentation. The fermentation process utilizes Clostridia bacteria that produce butanol, ethanol and acetone solvents at a total yield of 0.26 g/g sugars. A conditioning scheme is developed, which demonstrates that it is possible to utilize the hemicellulose sugars from this agricultural waste stream by traditional ABE fermentation. Fractionation as well as sugar hydrolysis in the spent liquor is hindered by the high cation content of OPEFB, which can be partly removed by acidic leaching suggesting that a better deashing method is necessary. Furthermore, it is inferred that better and more selective lignin removal is needed during conditioning to improve liquor fermentability.

Wheat flour based propionic acid fermentation: an economic approach.

A process for the fermentative production of propionic acid from whole wheat flour using starch and gluten as nutrients is presented. Hydrolysis of wheat flour starch using amylases was optimized. A batch fermentation of hydrolysate supplemented with various nitrogen sources using Propionibacterium acidipropionici NRRL B 3569 was performed. The maximum production of 48.61, 9.40, and 11.06 g of propionic acid, acetic acid and succinic acid, respectively, was found with wheat flour hydrolysate equivalent to 90 g/l glucose and supplemented with 15 g/l yeast extract. Further, replacement of yeast extract with wheat gluten hydrolysate showed utilization of gluten hydrolysate without compromising the yields and also improving the economics of the process. The process so developed could be useful for production of animal feed from whole wheat with in situ production of preservatives, and also suggest utilization of sprouted or germinated wheat for the production of organic acids.

Gellan Gum as Immobilization Matrix for Production of Cyclosporin A

N-acyl-homoserine lactone (AHL)-mediated quorum sensing (QS) plays a major role in development of biofilms, which contribute to rise in infections and biofouling in water-related industries. Interference in QS, called quorum quenching (QQ), has recieved a lot of attention in recent years. Rhodococcus spp. are known to have prominent quorum quenching activity and in previous reports it was suggested that this genus possesses multiple QQ enzymes, but only one gene, qsdA, which encodes an AHL-lactonase belonging to phosphotriesterase family, has been identified. Therefore, we conducted a whole genome sequencing and analysis of Rhodococcus sp. BH4 isolated from a wastewater treatment plant. The sequencing revealed another gene encoding a QQ enzyme (named jydB) that exhibited a high AHL degrading activity. This QQ enzyme had a 46% amino acid sequence similarity with the AHL-lactonase (AidH) of Ochrobactrum sp. T63. HPLC analysis and AHL restoration experiments by acidification revealed that the jydB gene encodes an AHL-lactonase which shares the known characteristics of the α/β hydrolase family. Purified recombinant JydB demonstrated a high hydrolytic activity against various AHLs. Kinetic analysis of JydB revealed a high catalytic efficiency (kcat/KM) against C4-HSL and 3-oxo-C6 HSL, ranging from 1.88 × 106 to 1.45 × 106 M-1 s-1, with distinctly low KM values (0.16 - 0.24 mM). This study affirms that the AHL degrading activity and biofilm inhibition ability of Rhodococcus sp. BH4 may be due to the presence of multiple quorum quenching enzymes, including two types of AHL-lactonases, in addition to AHL-acylase and oxidoreductase, for which the genes have yet to be described.