Water-immiscible Organic Solvent Research Articles

New Recombinant Cold-Adapted and Organic Solvent Tolerant Lipase from Psychrophilic Pseudomonas sp. LSK25, Isolated from Signy Island Antarctica.

In recent years, studies on psychrophilic lipases have become an emerging area of research in the field of enzymology. The study described here focuses on the cold-adapted organic solvent tolerant lipase strain Pseudomonas sp. LSK25 isolated from Signy Station, South Orkney Islands, maritime Antarctic. Strain LSK25 lipase was successfully cloned, sequenced, and over-expressed in an Escherichia coli system. Sequence analysis revealed that the lipase gene of Pseudomonas sp. LSK25 consists of 1432 bp, lacks an N-terminal signal peptide and encodes a mature protein consisting of 476 amino acids. The recombinant LSK25 lipase was purified by single-step purification using Ni-Sepharose affinity chromatography and had a molecular mass of approximately 65 kDa. The final recovery and purification fold were 44% and 1.3, respectively. The LSK25 lipase was optimally active at 30 °C and at pH 6. Stable lipolytic activity was reported between temperatures of 5–30 °C and at pH 6–8. A significant enhancement of lipolytic activity was observed in the presence of Ca2+ ions, the organic lipids of rice bran oil and coconut oil, a synthetic C12 ester and a wide range of water immiscible organic solvents. Overall, lipase strain LSK25 is a potentially desirable candidate for biotechnological application, due to its stability at low temperatures, across a range of pH and in organic solvents.

Model Amphiphilic Polymer Conetworks in Water: Prediction of Their Ability for Oil Solubilization.

In this work, we computationally explored the ability of water-swollen, model ionizable ABA triblock copolymer-based amphiphilic polymer conetworks (APCNs) to solubilize a water-immiscible organic solvent (oil), via Gibbs free energy minimization. This was done as a function of the conetwork hydrophobe (A-blocks) mol fraction and the degree of ionization of the hydrophilic B-blocks. Expectedly, highest oil solubilization capacities were calculated for the most hydrophobic and least ionized APCNs, which could absorb up to 6.4 times more oil than water and exhibited a lamellar morphology. Our results also included a phase diagram, which indicated transitions from spheres to cylinders, lamellae, and unimers in oil, as the hydrophobe content increased and the degree of ionization decreased. All of these transitions were accompanied by discontinuous changes in the degrees of swelling in the aqueous and oil nanophases, discontinuous changes in the asymmetry ratios (for the anisotropic morphologies), and discontinuous changes in the oil solubilization capacities. This is the first time that a dual discontinuous volume phase transition is reported within a polymer gel.

Biochemical profiles of two thermostable and organic solvent-tolerant esterases derived from a compost metagenome.

Owing to the functional versatility and potential applications in industry, interest in lipolytic enzymes tolerant to organic solvents is increasing. In this study, functional screening of a compost soil metagenome resulted in identification of two lipolytic genes, est1 and est2, encoding 270 and 389 amino acids, respectively. The two genes were heterologously expressed and characterized. Est1 and Est2 are thermostable enzymes with optimal enzyme activities at 80 and 70°C, respectively. A second-order rotatable design, which allows establishing the relationship between multiple variables with the obtained responses, was used to explore the combined effects of temperature and pH on esterase stability. The response curve indicated that Est1, and particularly Est2, retained high stability within a broad range of temperature and pH values. Furthermore, the effects of organic solvents on Est1 and Est2 activities and stabilities were assessed. Notably, Est2 activity was significantly enhanced (two- to tenfold) in the presence of ethanol, methanol, isopropanol, and 1-propanol over a concentration range between 6 and 30% (v/v). For the short-term stability (2h of incubation), Est2 exhibited high tolerance against 60% (v/v) of ethanol, methanol, isopropanol, DMSO, and acetone, while Est1 activity resisted these solvents only at lower concentrations (below 30%, v/v). Est2 also displayed high stability towards some water-immiscible organic solvents, such as ethyl acetate, diethyl ether, and toluene. With respect to long-term stability, Est2 retained most of its activity after 26days of incubation in the presence of 30% (v/v) ethanol, methanol, isopropanol, DMSO, or acetone. All of these features indicate that Est1 and Est2 possess application potential.

A comparative study of water-immiscible organic solvents in the production of furfural from xylose and birch hydrolysate

Furfural (FUR) was produced from xylose using a biphasic batch reaction system. Water-immiscible organic solvents such as isophorone, 2-methyltetrahydrofuran (2-MTHF) and cyclopentyl methyl ether (CPME) were used to promptly extract FUR from the aqueous phase in order to avoid the degradation to humins as largely as possible. The effect of time, temperature, organic solvent and organic-to-aqueous ratio on xylose conversion and FUR yield were investigated in auto-catalyzed conditions. Experiments at three temperatures (170, 190 and 210°C) were carried out in a stirred microwave-assisted batch reactor, which established the optimal conditions for achieving the highest FUR yield. The maximum FUR yields from xylose were 78mol% when using CPME, 48mol% using isophorone and 71mol% in the case of 2-MTHF at an aqueous to organic phase ratio of 1:1 (v/v). Birch hydrolysate was also used to show the high furfural yield that can be obtained in the biphasic system under optimized conditions. The present study suggests that CPME can be used as a green and efficient extraction solvent for the conversion of xylose into furfural without salt addition.

Two-phase micro-electromembrane extraction across free liquid membrane for determination of acidic drugs in complex samples

A dynamic two-phase micro-electromembrane extraction (μ-EME) using electrically induced transfer of charged analytes directly into free liquid membrane (FLM) is proposed as a novel technique for improving enrichment capabilities of μ-EME. The presented set-up employs aqueous sample as donor solution and water immiscible organic solvent (1-octanol) as FLM, which form the two-phase μ-EME system for efficient extraction of model acidic drugs (ibuprofen, ketoprofen, naproxen and diclofenac) from standard solutions, human urine, human serum and wastewater samples. The FLM eliminates migration of matrix components from the complex samples and simultaneously it acts as an acceptor solution for selective trapping and enrichment of the analyte ions. Electrodes are immersed directly into the sample and the FLM and replenishment of analyte ions at the sample/FLM phase interface is accomplished by stirring the sample solution using a conventional laboratory stirrer. At optimized two-phase μ-EME conditions (100 V, 15 min, 1000 rpm) and optimized volume ratio of sample to FLM (480:16 μL), extraction recoveries of 60–97% and enrichment factors up to 29.1 were achieved. Determination of the acidic drugs in resulting FLMs was achieved by capillary electrophoresis with ultraviolet detection with good linearity (r2 ≥ 0.9998) and low limits of detection (4–20 ng/mL).

Determination of Anions in Water-immiscible Organic Solvents by Ion Chromatography Combined with In-line Matrix Elimination

Determination of Anions in Water-immiscible Organic Solvents by Ion Chromatography Combined with In-line Matrix Elimination

Polyelectrolyte based technique for sequestration of protein from an aqueous phase to an organic solvent

Abstract Separation of proteins from a complex multi-component suspension of broken down cells is technically challenging. In this regard, a technique that is capable of distinguishing proteins from other major cellular components (such as carbohydrates, lipids, and other cellular debris) could immensely simplify downstream processing. In this work, we report the ability to use the inherent zwitterionic properties of proteins to latch amphiphilic cationic electrolytes on to protein surfaces so that proteins can be extracted from an aqueous suspension into a water-immiscible organic solvent. Here we show the separation of egg albumin (a model protein) from an aqueous suspension to adjacent hexane phase using poly-(diallyl dimethyl ammonium) chloride (PolyDADMAC). Results show that, under optimum pH and polyelectrolyte concentrations, proteins can be successfully migrated from water to the adjacent hexane phase. The amount and molecular weight of electrolyte, and solution pH played vital roles in determining the amount of protein that could be migrated. Low molecular weight polyelectrolyte under neutral pH conditions, low equilibration time and 1:2 hexane: water ratio favored highest protein separation (∼85% dry weight). FTIR studies indicated that the secondary structure of the protein was preserved after migration of the protein from the aqueous phase to solvent under the protection of the polyelectrolyte coating. The results suggest that ionic electrolytes with hydrophobic moieties could be used as molecular transport vehicles to safely separate proteins from a bulk aqueous phase to an immiscible solvent phase.

Cloning, overexpression, and characterization of a novel organic solvent-tolerant lipase from Paenibacillus pasadenensis CS0611

We found a novel lipase gene in the Paenibacillus pasadenensis CS0611 strain. The lipase gene sequence was cloned into the pET-28a expression vector to construct a recombinant lipase protein containing 6 × His tags at the C- and N-termini, respectively. High-level expression of the lipase in E.coli BL21 (DE3) was obtained upon induction with IPTG at 20 °C. The recombinant lipase activity was approximately 1631-fold higher than the wild type. His-tagged recombinant lipase was purified rapidly and efficiently by using Ni-charged affinity chromatography with 63.5% recovery and a purification factor of 10.78. The purified lipase was stable in a broad range of temperatures and pH values, with the optimal temperature and pH being 50 °C and 7.0, respectively. Its activity was stimulated to different degrees in the presence of metal ions such as Ca2+, Mg2+, and some non-ionic surfactants. In addition, the purified lipase was activated by a series of water-miscible organic solvents such as some short carbon chain alcohols and was highly tolerant to some water-immiscible organic solvents.

Equilibrium solubility measurement of compounds with low dissolution rate by Higuchi's Facilitated Dissolution Method. A validation study

Incubation time plays a critical role in the accurate measurement of equilibrium solubility of compounds. Substances which dissolve very slowly generally need long incubation times (days or weeks) to reach equilibrium. However, long times may pose several problems, such as decomposition of solute, molding of buffer, and drifting of pH. Higuchi in 1979 proposed the Facilitated Dissolution Method (FDM) to dramatically reduce incubation time. It employs a small volume of water-immiscible organic solvent to partly solubilize the sample and thereby increase the surface area available for dissolution. The method has been used only rarely. In this study we performed a systematic validation of FDM using progesterone as model compound. The reference solubility value, 7.95±0.21μg/mL (p<0.05, n=5), was determined in Britton-Robinson buffer solution (pH7.4) at 25.0°C by the standardized protocol of Saturation Shake-Flask (SSF) method. Also, the solubility was measured by the FDM approach under varied experimental conditions (e.g., type and volume of organic solvent, time of agitation, and amount of solid excess), and compared to the reference value. It was demonstrated that the small amount of organic solvent used in the FDM does not impact the measured solubility, compared to the reference value. Additionally, four compounds of low dissolution rate (dexamethasone, digoxin, haloperidol and cosalane) were used to demonstrate that FDM can reduce the long equilibration time to the standardized 24h (6h stirring and 18h sedimentation). The time dependence of solubility equilibrium was measured by SSF, and the results were compared with those obtained by FDM. Our study, based on >200 solubility experiments, supports the validity of Higuchi's method. In this study we propose a standardized protocol for the FDM, where 1% v/v of organic solvent is used. Octane (or isooctane) was found to be suitable for highly hydrophobic compounds. Alternatively, octanol or 1,2-dichloroethane can be used for less lipophilic compounds.

Novel thermostable amine transferases from hot spring metagenomes.

Hot spring metagenomes, prepared from samples collected at temperatures ranging from 55 to 95°C, were submitted to an in silico screening aimed at the identification of novel amine transaminases (ATAs), valuable biocatalysts for the preparation of optically pure amines. Three novel (S)-selective ATAs, namely Is3-TA, It6-TA, and B3-TA, were discovered in the metagenome of samples collected from hot springs in Iceland and in Italy, cloned from the corresponding metagenomic DNAs and overexpressed in recombinant form in E. coli. Functional characterization of the novel ATAs demonstrated that they all possess a thermophilic character and are capable of performing amine transfer reactions using a broad range of donor and acceptor substrates, thus suggesting a good potential for practical synthetic applications. In particular, the enzyme B3-TA revealed to be exceptionally thermostable, retaining 85% of activity after 5days of incubation at 80°C and more than 40% after 2weeks under the same condition. These results, which were in agreement with the estimation of an apparent melting temperature around 88°C, make B3-TA, to the best of our knowledge, the most thermostable natural ATA described to date. This biocatalyst showed also a good tolerance toward different water-miscible and water-immiscible organic solvents. A detailed inspection of the homology-based structural model of B3-TA showed that the overall active site architecture of mesophilic (S)-selective ATAs was mainly conserved in this hyperthermophilic homolog. Additionally, a subfamily of B3-TA-like transaminases, mostly uncharacterized and all from thermophilic microorganisms, was identified and analyzed in terms of phylogenetic relationships and sequence conservation.

Production of bioethanol by Zymomonas mobilis in high-gravity extractive fermentations

Continuous removal of ethanol during production by the anaerobic bacterium Zymomonas mobilis mitigated ethanol inhibition. Ethanol inhibited its own production at a concentration of >20gL−1. Ethanol productivity was enhanced by continuous in situ extraction with water-immiscible organic solvents. Several solvents were evaluated for biocompatibility with Z. mobilis and the ability to extract ethanol from a batch fermentation. In situ batch extractive fermentations were carried out in a 2-L stirred bioreactor (1:1 volume ratio of solvent and fermentation broth) with an optimal glucose concentration of 150gL−1, 35°C, and an agitation speed of 150rpm. Oleyl alcohol, iso-octadecanol and 2-octyl-1-dodecanol were the three solvents that were most compatible with Z. mobilis. With these solvents, the cell viability relative to control (no solvent) was 1.48±0.40, 1.03±0.18 and 1.05±0.07, respectively, after two days of exposure. All solvents tested improved the final biomass concentration, the glucose consumption and ethanol production relative to control, but iso-octadecanol was clearly the most effective solvent. Using iso-octadecanol, the maximum ethanol concentration of 75±3gL−1 was attained. This was nearly 1.25-fold that of the control fermentation. For this solvent, the ethanol yield on glucose was 0.485±0.005gg−1 compared to a yield of 0.468±0.005gg−1 for the control culture.

Integration of the free liquid membrane into electrokinetic supercharging – capillary electrophoresis for the determination of cationic herbicides in environmental water samples

A new approach based on the integration of the free liquid membrane (FLM) into electrokinetic supercharging (EKS) was demonstrated to be a new powerful tool used in order to enhance online preconcentration efficiency in capillary electrophoresis (CE). A small plug of water immiscible organic solvent was used as a membrane interface during the electrokinetic sample injection step in EKS in order to significantly enhance the analyte stacking efficiency. The new online preconcentration strategy was evaluated for the determination of paraquat and diquat present in the environmental water samples. The optimised FLM-EKS conditions employed were as follows: hydrodynamic injection (HI) of 20mM potassium chloride as leading electrolyte at 50mbar for 75s (3% of the total capillary volume) followed by the HI of tris(2-ethylhexyl) phosphate (TEHP) as FLM at a 1mm length (0.1% of the capillary volume). The sample was injected at 10kV for 360s, followed by the HI of 20mM cetyl trimethylammonium bromide (CTAB) as terminating electrolyte at 50mbar for 50s (2% of the total capillary volume). The separation was performed in 12mM ammonium acetate and 30mM NaCl containing 20% MeOH at +25kV with UV detection at 205nm. Under optimised conditions, the sensitivity was enhanced between 1500- and 1866-fold when compared with the typical HI at 50mbar for 50s. The detection limit of the method for paraquat and diquat was 0.15–0.20ng/mL, with RSDs below 5.5%. Relative recoveries in spiked river water were in the range of 95.4–97.5%. A comparison was also made between the proposed approach with sole preconcentration of the field-enhanced sample injection (FASI) and EKS in the absence of the FLM.

Kinetic resolution of racemic styrene oxide at a high concentration by recombinant Aspergillus usamii epoxide hydrolase in an n-hexanol/buffer biphasic system

Using the cell-free extract of engineered E. coli/Aueh2, expressing the recombinant Aspergillus usamii epoxide hydrolase (reAuEH2), as a biocatalyst, the kinetic resolution technique of racemic styrene oxide (rac-SO) was examined. In a phosphate buffer system (50mM, pH 7.0), 200mM rac-SO was efficiently resolved, obtaining (S)-SO with 98.1% enantiomeric excess (e.e.), whereas (S)-SO only with 45.2% e.e. was obtained from 750mM rac-SO. The analytical results verified that reAuEH2 shows tolerance towards high substrate concentration but is inactivated at a product concentration of 300mM. To produce (S)-SO with the high concentration, e.e. and volumetric productivity, n-hexanol was selected from a variety of water-miscible and water-immiscible organic solvents to construct an n-hexanol/buffer biphasic system. The optimal phase volume ratio, substrate over enzyme ratio and temperature were 1:1 (v/v), 6:1 (w/w) and 25°C, respectively. In an optimized biocatalytic system, a gram-scale resolution of rac-SO at a high concentration of 1M (120g/L) was performed at 25°C for 2h, obtaining (S)-SO with 98.2% e.e., 34.3% yield (maximum yield of 50%). The substrate concentration and volumetric productivity (1M, 20.6g/L/h) in a biphasic system significantly increased compared with those (0.2M, 3.1g/L/h) in a phosphate buffer system. The efficient resolution of rac-SO at a high concentration in a biphasic system makes it a promising technique for preparing a highly value-added enantiopure (S)-SO with high volumetric productivity.

An effective method for the extraction and purification of chlorogenic acid from ramie (Boehmeria nivea L.) leaves using acidic ionic liquids

As a green solvent, ionic liquid was developed as a substitute for conventional volatile organic solvents in the extraction field. In this work, several task-specific acidic ionic liquids (TAILs) were applied for ultrasonic-assisted extraction (UAE) of chlorogenic acid (CGA) from ramie leaves. The influencing parameters, viz. ionic liquid (IL) concentration, liquid–solid ratio, and ultrasonic time, were optimized using response surface methodology (RSM). Of the ILs examined, 1-butyl-3-methylimidazolium hydrogen sulfate ([C4mim]HSO4) exhibited the best extraction ability. The RSM results revealed that optimized conditions are IL concentration 52.28% (w/w), liquid–solid ratio 89.05:1, and ultrasonic time 49.88min. An aqueous two-phase system (ATPS) formed by IL extract and salt was then developed for further extraction and purification of CGA. The influencing factors of salt type and concentration, pH, and temperature were investigated in detail. The maximum extraction efficiency of 96.18% was obtained in the IL-rich phase under the conditions of 35.83% (w/w) [C4mim]HSO4 and 14.13% (w/w) Na2SO4, pH 3 and temperature 37°C. After extraction, the isolation of CGA and recycling of IL were achieved by back-extraction using a water immiscible organic solvent. This simple and efficient IL-UAE coupled with ATPS has potentiality for the extraction and purification of CGA and other compounds in pilot- or large-scale operations.

Study on the disparate transition behaviors of the electrical/physical properties in PEDOT:PSS film depending on solvent species under a follow-up solution-treatment process

In most solution-processed organic devices, a poly(3,4-ethylenedioxythiophene) (PEDOT) polymerized with poly(4-styrenesulfonate) (PSS) film is inevitably affected by various conditions during the subsequent solution-coating processes. To investigate the effects of direct solvent exposure on the properties of PEDOT polymerized with PSS (PEDOT:PSS) films, photoemission spectroscopy-based analytical methods were used before and after solvent-coating processes. Our results clearly indicate that PEDOT:PSS films undergo a different transition mechanism depending on the solubility of the solvent in water. The water-miscible solvents induce the solvation of hydrophilic PSS chains. As a result, this process allows the solvent to diffuse into the PEDOT:PSS film, and a conformational change between PEDOT and PSS occurs. On the other hand, the water-immiscible organic solvents cause the partial adsorption of solvent molecules at the PE surface, which leads to changes in the surface properties, including work function. Based on our finding, we demonstrate that the energy-level alignments at the organic semiconductor/electrode interface for the PEDOT:PSS films can be controlled by simple solvent treatments.

Markedly improving asymmetric oxidation of 1-(4-methoxyphenyl) ethanol with Acetobacter sp. CCTCC M209061 cells by adding deep eutectic solvent in a two-phase system.

BackgroundEnantiopure (S)-1-(4-methoxyphenyl) ethanol {(S)-MOPE} can be employed as an important synthon for the synthesis of cycloalkyl [b] indoles with the treatment function for general allergic response. To date, the biocatalytic resolution of racemic MOPE through asymmetric oxidation in the biphasic system has remained largely unexplored. Additionally, deep eutectic solvents (DESs), as a new class of promising green solvents, have recently gained increasing attention in biocatalysis for their excellent properties and many successful examples in biocatalytic processes. In this study, the biocatalytic asymmetric oxidation of MOPE to get (S)-MOPE using Acetobacter sp. CCTCC M209061 cells was investigated in different two-phase systems, and adding DES in a biphasic system was also explored to further improve the reaction efficiency of the biocatalytic oxidation.ResultsOf all the examined water-immiscible organic solvents and ionic liquids (ILs), 1-butyl-3-methylimidazolium hexafluorophoshpate ([C4MIM][PF6]) afforded the best results, and consequently was selected as the second phase of a two-phase system for the asymmetric oxidation of MOPE with immobilized Acetobacter sp. CCTCC M209061 cells. For the reaction performed in the [C4MIM][PF6]/buffer biphasic system, under the optimized conditions, the initial reaction rate, the maximum conversion and the residual substrate e.e. recorded 97.8 μmol/min, 50.5 and >99.9 % after 10 h reaction. Furthermore, adding the DES [ChCl][Gly] (10 %, v/v) to the aqueous phase, the efficiency of the biocatalytic oxidation was rose markedly. The optimal substrate concentration and the initial reaction rate were significantly increased to 80 mmol/L and 124.0 μmol/min, respectively, and the reaction time was shortened to 7 h with 51.3 % conversion. The immobilized cell still retained over 72 % of its initial activity after 9 batches of successive reuse in the [C4MIM][PF6]/[ChCl][Gly]-containing buffer system. Additionally, the efficient biocatalytic process was feasible up to a 500-mL preparative scale.ConclusionThe biocatalytic asymmetric oxidation of MOPE with Acetobacter sp. CCTCC M209061 cells was successfully conducted in the [C4MIM][PF6]-containing biphasic system with high conversion and enantioselectivity, and the reaction efficiency was further enhanced by adding [ChCl][Gly] to the reaction system. The efficient biocatalytic process was promising for the preparation of enantiopure (S)-MOPE.

Effect of Lyophilization Technology on Dried Carnation Flower

The study was undertaken to explore the effect of lyophilization technology on dried carnation flower. Lyophilization helped to retain the natural colour, form and texture of the dried flowers. Certain preservatives were selected, tested and evaluated to keep hold of innate characteristics of flowers. The flower treated with silver thiosulphate (STS) was found to be effective hydration treatments resulting flowers remained fresh up to 9d. Floral preservative (T8) was the best composition which retains inherent qualities of the flowers. The water content of the plant was replaced with a water immiscible organic solvent for pre treatments. Application of glazing medium as sealant was found best to protect flower from external environment conditions.

Graphene oxide-based dispersive solid-phase extraction combined with in situ derivatization and gas chromatography–mass spectrometry for the determination of acidic pharmaceuticals in water

A fast and low-cost sample preparation method of graphene based dispersive solid-phase extraction combined with gas chromatography–mass spectrometric (GC–MS) analysis, was developed. The procedure involves an initial extraction with water-immiscible organic solvent, followed by a rapid clean-up using amine functionalized reduced graphene oxide as sorbent. Simple and fast one-step in situ derivatization using trimethylphenylammonium hydroxide was subsequently applied on acidic pharmaceuticals serving as model analytes, ibuprofen, gemfibrozil, naproxen, ketoprofen and diclofenac, before GC–MS analysis. Extraction parameters affecting the derivatization and extraction efficiency such as volume of derivatization agent, effect of desorption solvent, effect of pH and effect of ionic strength were investigated. Under the optimum conditions, the method demonstrated good limits of detection ranging from 1 to 16ngL−1, linearity (from 0.01 to 50 and 0.05 to 50μgL−1, depending on the analytes) and satisfactory repeatability of extractions (relative standard deviations, below 13%, n=3).

Enhancement of Palmarumycin C12 and C13 Production by the Endophytic Fungus Berkleasmium sp. Dzf12 in an Aqueous-Organic Solvent System.

The endophytic fungus Berkleasmium sp. Dzf12, isolated from Dioscorea zingiberensis, was found to produce palmarumycins C12 and C13 which possess a great variety of biological activities. Seven biocompatible water-immiscible organic solvents including n-dodecane, n-hexadecane, 1-hexadecene, liquid paraffin, dibutyl phthalate, butyl oleate and oleic acid were evaluated to improve palmarumycins C12 and C13 production in suspension culture of Berkleasmium sp. Dzf12. Among the chosen solvents both butyl oleate and liquid paraffin were the most effective to improve palmarumycins C12 and C13 production. The addition of dibutyl phthalate, butyl oleate and oleic acid to the cultures of Berkleasmium sp. Dzf12 significantly enhanced palmarumycin C12 production by adsorbing palmarumycin C12 into the organic phase. When butyl oleate was fed at 5% (v/v) in medium at the beginning of fermentation (day 0), the highest palmarumycin C12 yield (191.6 mg/L) was achieved, about a 34.87-fold increase in comparison with the control (5.3 mg/L). n-Dodecane, 1-hexadecene and liquid paraffin had a great influence on the production of palmarumycin C13. When liquid paraffin was added at 10% (v/v) in medium on day 3 of fermentation, the palmarumycin C13 yield reached a maximum value (134.1 mg/L), which was 4.35-fold that of the control (30.8 mg/L). Application of the aqueous-organic solvent system should be a simple and efficient process strategy for enhancing palmarumycin C12 and C13 production in liquid cultures of the endophytic fungus Berkleasmium sp. Dzf12.

Repulsive effect of salt on solvent extraction of 2,3-butanediol from aqueous fermentation solution

BACKGROUND Solvent extraction of 2,3-butanediol from dilute fermented systems is a challenging downstream unit operation. Addition of certain kosmotropic salts to the aqueous fermentation broth during the solvent extraction process can have considerable influence on the distribution of 2,3-butanediol. This work evaluates the usefulness of solvent extraction and the salting-out effect on the extraction characteristic of 2,3-butanediol in aqueous synthetic and fermented solutions. RESULTS Addition of 10 wt% dipotassium phosphate (K2HPO4) to the fermented 2,3-butanediol broth during solvent extraction results in 1.5–3-fold increase in distribution coefficient (Kd) and 7–14-fold increase in separation factor (α) for 2,3-butanediol using 2-sec butyl phenol and 1-butanol as extractants. Of various solvent–salt combinations studied; 2-sec butyl phenol + 10 wt% K2HPO4 provided Kd and α of 3.04 and 59.17 whereas 1-butanol + 10 wt% K2HPO4 provided Kd and α of 2.41 and 15.68, respectively, in fermented 2,3-butanediol broth solution. CONCLUSION This study demonstrated the prospects of repulsive salts addition to the fermented broth during the solvent extraction process. The results showed that this study may be used as a model to choose an appropriate combination of water-immiscible (hydrophobic) organic solvent and inorganic salt type and concentration for the economical recovery of 2,3-butanediol from fermentation broth. © 2014 Society of Chemical Industry