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Paul Christakopoulos - One of the best experts on this subject based on the ideXlab platform.
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the feruloyl esterase system of talaromyces stipitatus determining the hydrolytic and synthetic specificity of tsfaec
Journal of Biotechnology, 2006Co-Authors: Christina Vafiadi, Evangelos Topakas, Paul ChristakopoulosAbstract:The active site of the recombinant Talaromyces stipitatus type-C feruloyl esterase (TsFaeC) was probed using a series of C1–C4 alkyl ferulates and methyl esters of phenylalkanoic and cinnamic acids. The enzyme was active on 23 of the 34 substrates tested. Lengthening or shortening the aliphatic side chain while maintaining the same aromatic substitutions completely abolished the enzyme activity. Maintaining the phenylpropenoate structure but altering the substitutions of the aromatic ring demonstrated the importance of hydroxyl groups on meta and/or para position of the benzoic ring. The highest catalytic efficiency of TsFaeC for methyl cinnamates was shown on methyl 3,4-dihydroxy cinnamate and on its hydro form (3,4-dihydroxy-phenyl-propionate). Maintaining the ferulate structure but altering the esterified alkyl group, the comparison of kcat and kcat/Km values showed that the enzyme hydrolysed faster and more efficiently than ethyl ferulate. Alkyl ferulates were applied also for substrate selectivity mapping of feruloyl esterase to catalyze feruloyl group transfer to l-arabinose, using as a reaction system a ternary water–Organic Mixture consisting of n-hexane, t-butanol and water. The reaction parameters affecting the feruloylation rate and the conversion of the enzymatic synthesis, such as the composition of the reaction media, temperature, substrate and enzyme concentration have been investigated.
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sporotrichum thermophile type c feruloyl esterase stfaec purification characterization and its use for phenolic acid sugar ester synthesis
Enzyme and Microbial Technology, 2005Co-Authors: Evangelos Topakas, Haralambos Stamatis, Christina Vafiadi, Paul ChristakopoulosAbstract:Abstract A feruloyl esterase (StFaeC) produced by Sporotrichum thermophile was purified to homogeneity. The native StFaeC was homodimer with a subunit of M r 23,000 and pI 3.1. The enzyme activity was optimal at pH 6.0 and 55 °C. The esterase displayed remarkable stability at pH 10.0 and retained 50% of its activity after 133 and 55 min at 55 and 60 °C, respectively. Determination of k cat / K m revealed that the enzyme had a broad spectrum of activity against the (hydroxyl) cinnamate esters indicating a type C feruloyl esterase. The enzyme was active on substrates containing ferulic acid ester linked to the C-5 and C-2 linkages of arabinofuranose and hydrolysed 4-nitrophenyl-5- O - trans -feruloyl-α- l -arabinofuranoside three times more efficiently than 4-nitrophenyl-2- O - trans -feruloyl-α- l -arabinofuranoside. Ferulic acid was efficiently released from wheat bran when the esterase was incubated together with xylanase from S. thermophile (a maximum of 41% total ferulic acid released after 1 h incubation). StFaeC by itself could release FA but at a level almost 10-fold lower than that obtained in the presence of xylanase. The potential of StFaeC for the synthesis of various phenolic acid esters was examined using as a reaction system a ternary water–Organic Mixture consisting of n-hexane, 1-butanol and water. Also StFaeC catalyzed the transfer of the feruloyl group to l -arabinose in a similar system using t -butanol, with about a 40% conversion of l -arabinose to feruloylated derivative was achieved. This work is the first example of enzymatic feruloylation of a carbohydrate.
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mapping the hydrolytic and synthetic selectivity of a type c feruloyl esterase stfaec from sporotrichum thermophile using alkyl ferulates
Tetrahedron-asymmetry, 2005Co-Authors: Christina Vafiadi, Ian D. Suckling, Evangelos Topakas, Ken K. Y. Wong, Paul ChristakopoulosAbstract:Abstract The active site of Sporotrichum thermophile type C feruloyl esterase (StFaeC) was probed using a series of C1–C4 alkyl ferulates. The affinities for straight and branched alkyl ferulates were demonstrated by the Km values of 1.64–0.51 and 0.19–0.1, respectively. Comparison of kcat and kcat/Km values shows that the enzyme hydrolyzed n-propyl ferulate faster and iso-propyl ferulate more efficiently. Alkyl ferulates were applied also for substrate selectivity mapping of feruloyl esterase to catalyze feruloyl group transfer to l -arabinose, using as a reaction system a ternary water–Organic Mixture consisting of n-hexane, t-butanol and water. Lengthening the aliphatic side chain was the most significant factor causing lower synthetic activity of the enzyme. The reaction parameters affecting the feruloylation rate and the conversion of the enzymatic process, such as the temperature and substrate concentration have been investigated. Under identical reaction conditions, the enzyme feruloylated other monosaccharides such as d -arabinose, d -glucose, d -xylose, d -mannose, d -fructose, d -galactose, d -ribose and model substrates such as 4-nitrophenyl α- l -arabinofuranoside and 4-nitrophenyl α- l -arabinopyranoside.
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purification and characterization of a type b feruloyl esterase stfae a from the thermophilic fungus sporotrichum thermophile
Applied Microbiology and Biotechnology, 2004Co-Authors: Evangelos Topakas, Haralambos Stamatis, Paul Christakopoulos, Peter BielyAbstract:A feruloyl esterase (StFAE-A) produced by Sporotrichum thermophile was purified to homogeneity. The purified homogeneous preparation of native StFAE-A exhibited a molecular mass of 57.0±1.5 kDa, with a mass of 33±1 kDa on SDS-PAGE. The pI of the enzyme was estimated by cation-exchange chromatofocusing to be at pH 3.1. The enzyme activity was optimal at pH 6.0 and 55–60 °C. The purified esterase was stable at the pH range 5.0–7.0. The enzyme retained 70% of activity after 7 h at 50 °C and lost 50% of its activity after 45 min at 55 °C and after 12 min at 60 °C. Determination of kcat/Km revealed that the enzyme hydrolyzed methyl p-coumarate 2.5- and 12-fold more efficiently than methyl caffeate and methyl ferulate, respectively. No activity on methyl sinapinate was detected. The enzyme was active on substrates containing ferulic acid ester linked to the C-5 and C-2 linkages of arabinofuranose and it hydrolyzed 4-nitrophenyl 5-O-trans-feruloyl-α-l-arabinofuranoside (NPh-5-Fe-Araf) 2-fold more efficiently than NPh-2-Fe-Araf. Ferulic acid (FA) was efficiently released from destarched wheat bran when the esterase was incubated together with xylanase from S. thermophile (a maximum of 34% total ferulic acid released after 1 h incubation). StFAE-A by itself could release FA, but at a level almost 47-fold lower than that obtained in the presence of xylanase. The potential of StFAE-A for the synthesis of various phenolic acid esters was tested using a ternary water-Organic Mixture consisting of n-hexane, 1-butanol and water as a reaction system.
Kazuhiko Tsukagoshi - One of the best experts on this subject based on the ideXlab platform.
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Specific microfluidic behavior of ternary mixed carrier solvents of water–acetonitrile–ethyl acetate in open-tubular capillary chromatography and the chromatograms
Analytical Methods, 2012Co-Authors: Satoshi Fujinaga, Katsuya Unesaki, Shigeru Negi, Masahiko Hashimoto, Kazuhiko TsukagoshiAbstract:We have developed a capillary chromatography system using an open-tubular capillary tube and a ternary water–hydrophilic–hydrophobic Organic Mixture carrier solution. We call the system “tube radial distribution chromatography (TRDC) system”. In this study, fluorescence photographs of the dyes dissolved in the ternary mixed carrier solvents in the TRDC were examined under various conditions concerning the inner diameters, tube temperatures, tube effective lengths, and flow rates (or average linear velocities). The TRDC system used an untreated fused-silica capillary tube and a ternary water–acetonitrile–ethyl acetate Mixture carrier solution (3 : 8 : 4 volume ratio; the Organic solvent-rich solution). The tube radial distribution of the carrier solvents that generated the Organic solvent-rich major inner phase and the water rich minor outer phase could be observed in 50–700 μm inner diameters of the tubes at an average linear velocity of 11 cm min−1 through the fluorescence microscope–CCD camera. The model analyte solution of 1-naphthol, 1-naphthalenesulfonic acid, 2,6-naphthalenedisulfonic acid, and 1,3,6-naphthalenetrisulfonic acid was subjected to TRDC. They were separated and detected in this order with 50, 75, and 100 μm inner diameters of the open-tubular capillaries.
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fluorescence observation supporting capillary chromatography based on tube radial distribution of carrier solvents under laminar flow conditions
Analyst, 2011Co-Authors: Naoya Jinno, Masahiko Hashimoto, Mari Murakami, Kiyoshi Mizohata, Kazuhiko TsukagoshiAbstract:We developed a capillary chromatography system by using an open capillary tube made of fused-silica, polyethylene, or polytetrafluoroethylene, and a water–hydrophilic/hydrophobic Organic Mixture carrier solution, called tube radial distribution chromatography (TRDC) system. By comparing with chromatograms obtained via the TRDC system, fluorescence photographs and profiles of the fluorescent dyes dissolved in the carrier solvents in capillary tubes were observed under laminar flow conditions. The chromatograms were obtained for a model Mixture analyte consisting of 1-naphthol and 2,6-naphthalenedisulfonic acid with the TRDC system, by using a fused-silica capillary tube and a water–acetonitrile–ethyl acetate carrier solution. By altering the carrier flow rates, we examined the fluorescence photographs and profiles of the dyes, perylene and Eosin Y, dissolved in the carrier solvents in the capillary tube by using a fluorescence microscope equipped with a CCD camera. As confirmed by fluorescence observations, the major inner and minor outer phases generated in the capillary tube were based on the tube's radial distribution of the carrier solvents. We designed and manufactured a microreactor incorporating microchannels in which three narrow channels combined to form one wide channel. When the carrier solvents containing the dyes were fed into the channels, the inner and outer phase generations were also observed in the narrow and wide channels, strongly supporting the conclusions concerning the tube radial distribution phenomenon of the solvents.
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analytical conditions and separation performance of capillary chromatography based on the tube radial distribution of aqueous Organic Mixture carrier solvents under laminar flow conditions
Analytical Sciences, 2010Co-Authors: Naoya Jinno, Masahiko Hashimoto, Mari Murakami, Kazuhiko TsukagoshiAbstract:We have developed a capillary chromatography system using an open capillary tube made of fused-silica, polyethylene, or poly(tetrafluoroethylene), and a water–hydrophilic–hydrophobic Organic Mixture carrier solution. This tube radial distribution chromatography (TRDC) system works under laminar-flow conditions. In this study, the following analytical conditions in the TRDC system using a fused-silica capillary tube and a water–acetonitrile–ethyl acetate Mixture carrier solution were for the first time examined: tube temperature, 5 − 25°C; tube inner diameter, 50 − 250 μm; tube effective length, 100 − 200 cm; and flow rate, 0.2 − 1.5 μL min−1. For example, the effects of temperature on the separation performance in the TRDC system were observed with an Organic solvent-rich carrier solution; 1-naphthol and 2,6-naphthalenedisulfonic acid in a model Mixture were eluted with baseline separation over the temperature range of 5 − 23°C. The resolution, theoretical plate number, and height equivalent to the theoretical plate were calculated from the experimental data obtained by examining the effects of the tube length. A Mixture of 1-naphthol, Eosin Y, 1-naphthalenesulfonic acid, 2,6-naphthalenedisulfonic acid, and 1,3,6-naphthalenetrisulfonic acid was subjected to the present TRDC system, and the analytes in the Mixture solution were eluted in this order with the Organic solvent-rich carrier solution, providing good separation performance on the chromatogram.
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elution behavior of proteins in capillary chromatography using an untreated fused silica capillary tube and a water hydrophilic hydrophobic Organic Mixture carrier solvent
Chemistry Letters, 2010Co-Authors: Yuji Masuhara, Masahiko Hashimoto, Naoya Jinno, Kazuhiko TsukagoshiAbstract:We examined the elution behavior of isoluminol isothiocyanate (ILITC)-labeled proteins (alcohol dehydrogenase (ADH) and bovine serum albumin (BSA)) in a capillary chromatography system using an unt...
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Capillary chromatography based on tube radial distribution of aqueous-Organic Mixture carrier solvents.
Talanta, 2009Co-Authors: Naoya Jinno, Masahiko Hashimoto, Minoru Itano, Kazuhiko TsukagoshiAbstract:Abstract A capillary chromatography system was developed using open capillary tubes made of fused-silica, polyethylene, or poly(tetrafluoroethylene), and an aqueous–Organic Mixture (water–acetonitrile–ethyl acetate Mixture) as a carrier solution. Model analyte Mixture solutions, such as 2,6-naphthalenedisulfonic acid and 1-naphthol, Eosin Y and perylene, bis[N,N-bis(carboxymethyl)aminomethyl]fluorescein and 1,1’-bi-2-naphthol, and 2,7-naphthalenedisulfonic acid and p-nitroaniline, were injected into the capillary tube by a gravity method. The analyte solutions were subsequently delivered through the capillary tube with the carrier solution by a micro-syringe pump. The system worked under laminar flow conditions. The analytes were separated through the capillary tube and detected on-capillary by an absorption detector. For example, 2,6-naphthalenedisulfonic acid and 1-naphthol were detected in this order with a carrier solution of water–acetonitrile–ethyl acetate (volume ratio 15:3:2), while they were detected in the reverse order with a carrier solution of water–acetonitrile–ethyl acetate (volume ratio 2:9:4). The other analyte solutions were similarly separated by the system. The elution times of the analytes could be easily reversed by changing the component ratio of the solvents in the carrier solution.
Javier Fontalvo - One of the best experts on this subject based on the ideXlab platform.
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Liquid–Liquid Equilibrium and Molecular Toxicity of Active and Inert Diluents of the Organic Mixture Tri-iso-octylamine/Dodecanol/Dodecane as a Potential Liquid Membrane for Lactic Acid Removal
Journal of Chemical & Engineering Data, 2019Co-Authors: Alan D. Pérez, Verónica M. Gómez, Sneyder Rodríguez-barona, Javier FontalvoAbstract:Lactic acid can be in situ removed from a fermentation broth through reactive liquid extraction or a liquid membrane to enhance the fermentation process. The Organic Mixture tri-iso-octylamine (TiOA)/dodecanol/dodecane at 10 vol % of the amine is a potential Organic Mixture for lactic acid removal. Liquid–liquid equilibria with lactic acid aqueous solutions and molecular toxicity on the bacteria Lactobacillus casei ATCC 393 were measured with several dodecanol proportions in dodecane (0 to 90 vol %) and 10 vol % TiOA as potential solvents or membrane phases for LA removal from a fermentation broth. Effects of the Organic phase on the bacteria as cell growth, biomass production, glucose consumption, productivity, and product to biomass yield are analyzed. Dodecanol increases the lactic acid chemical equilibrium constant for the liquid–liquid equilibria, while increasing the molecular toxicity on the bacteria. However, for dodecanol concentrations from 30 to 40 vol % the value of the chemical equilibrium co...
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liquid liquid equilibrium and molecular toxicity of active and inert diluents of the Organic Mixture tri iso octylamine dodecanol dodecane as a potential liquid membrane for lactic acid removal
Journal of Chemical & Engineering Data, 2019Co-Authors: Alan D. Pérez, Verónica M. Gómez, Sneyder Rodriguezbarona, Javier FontalvoAbstract:Lactic acid can be in situ removed from a fermentation broth through reactive liquid extraction or a liquid membrane to enhance the fermentation process. The Organic Mixture tri-iso-octylamine (TiOA)/dodecanol/dodecane at 10 vol % of the amine is a potential Organic Mixture for lactic acid removal. Liquid–liquid equilibria with lactic acid aqueous solutions and molecular toxicity on the bacteria Lactobacillus casei ATCC 393 were measured with several dodecanol proportions in dodecane (0 to 90 vol %) and 10 vol % TiOA as potential solvents or membrane phases for LA removal from a fermentation broth. Effects of the Organic phase on the bacteria as cell growth, biomass production, glucose consumption, productivity, and product to biomass yield are analyzed. Dodecanol increases the lactic acid chemical equilibrium constant for the liquid–liquid equilibria, while increasing the molecular toxicity on the bacteria. However, for dodecanol concentrations from 30 to 40 vol % the value of the chemical equilibrium co...
Evangelos Topakas - One of the best experts on this subject based on the ideXlab platform.
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the feruloyl esterase system of talaromyces stipitatus determining the hydrolytic and synthetic specificity of tsfaec
Journal of Biotechnology, 2006Co-Authors: Christina Vafiadi, Evangelos Topakas, Paul ChristakopoulosAbstract:The active site of the recombinant Talaromyces stipitatus type-C feruloyl esterase (TsFaeC) was probed using a series of C1–C4 alkyl ferulates and methyl esters of phenylalkanoic and cinnamic acids. The enzyme was active on 23 of the 34 substrates tested. Lengthening or shortening the aliphatic side chain while maintaining the same aromatic substitutions completely abolished the enzyme activity. Maintaining the phenylpropenoate structure but altering the substitutions of the aromatic ring demonstrated the importance of hydroxyl groups on meta and/or para position of the benzoic ring. The highest catalytic efficiency of TsFaeC for methyl cinnamates was shown on methyl 3,4-dihydroxy cinnamate and on its hydro form (3,4-dihydroxy-phenyl-propionate). Maintaining the ferulate structure but altering the esterified alkyl group, the comparison of kcat and kcat/Km values showed that the enzyme hydrolysed faster and more efficiently than ethyl ferulate. Alkyl ferulates were applied also for substrate selectivity mapping of feruloyl esterase to catalyze feruloyl group transfer to l-arabinose, using as a reaction system a ternary water–Organic Mixture consisting of n-hexane, t-butanol and water. The reaction parameters affecting the feruloylation rate and the conversion of the enzymatic synthesis, such as the composition of the reaction media, temperature, substrate and enzyme concentration have been investigated.
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sporotrichum thermophile type c feruloyl esterase stfaec purification characterization and its use for phenolic acid sugar ester synthesis
Enzyme and Microbial Technology, 2005Co-Authors: Evangelos Topakas, Haralambos Stamatis, Christina Vafiadi, Paul ChristakopoulosAbstract:Abstract A feruloyl esterase (StFaeC) produced by Sporotrichum thermophile was purified to homogeneity. The native StFaeC was homodimer with a subunit of M r 23,000 and pI 3.1. The enzyme activity was optimal at pH 6.0 and 55 °C. The esterase displayed remarkable stability at pH 10.0 and retained 50% of its activity after 133 and 55 min at 55 and 60 °C, respectively. Determination of k cat / K m revealed that the enzyme had a broad spectrum of activity against the (hydroxyl) cinnamate esters indicating a type C feruloyl esterase. The enzyme was active on substrates containing ferulic acid ester linked to the C-5 and C-2 linkages of arabinofuranose and hydrolysed 4-nitrophenyl-5- O - trans -feruloyl-α- l -arabinofuranoside three times more efficiently than 4-nitrophenyl-2- O - trans -feruloyl-α- l -arabinofuranoside. Ferulic acid was efficiently released from wheat bran when the esterase was incubated together with xylanase from S. thermophile (a maximum of 41% total ferulic acid released after 1 h incubation). StFaeC by itself could release FA but at a level almost 10-fold lower than that obtained in the presence of xylanase. The potential of StFaeC for the synthesis of various phenolic acid esters was examined using as a reaction system a ternary water–Organic Mixture consisting of n-hexane, 1-butanol and water. Also StFaeC catalyzed the transfer of the feruloyl group to l -arabinose in a similar system using t -butanol, with about a 40% conversion of l -arabinose to feruloylated derivative was achieved. This work is the first example of enzymatic feruloylation of a carbohydrate.
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mapping the hydrolytic and synthetic selectivity of a type c feruloyl esterase stfaec from sporotrichum thermophile using alkyl ferulates
Tetrahedron-asymmetry, 2005Co-Authors: Christina Vafiadi, Ian D. Suckling, Evangelos Topakas, Ken K. Y. Wong, Paul ChristakopoulosAbstract:Abstract The active site of Sporotrichum thermophile type C feruloyl esterase (StFaeC) was probed using a series of C1–C4 alkyl ferulates. The affinities for straight and branched alkyl ferulates were demonstrated by the Km values of 1.64–0.51 and 0.19–0.1, respectively. Comparison of kcat and kcat/Km values shows that the enzyme hydrolyzed n-propyl ferulate faster and iso-propyl ferulate more efficiently. Alkyl ferulates were applied also for substrate selectivity mapping of feruloyl esterase to catalyze feruloyl group transfer to l -arabinose, using as a reaction system a ternary water–Organic Mixture consisting of n-hexane, t-butanol and water. Lengthening the aliphatic side chain was the most significant factor causing lower synthetic activity of the enzyme. The reaction parameters affecting the feruloylation rate and the conversion of the enzymatic process, such as the temperature and substrate concentration have been investigated. Under identical reaction conditions, the enzyme feruloylated other monosaccharides such as d -arabinose, d -glucose, d -xylose, d -mannose, d -fructose, d -galactose, d -ribose and model substrates such as 4-nitrophenyl α- l -arabinofuranoside and 4-nitrophenyl α- l -arabinopyranoside.
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purification and characterization of a type b feruloyl esterase stfae a from the thermophilic fungus sporotrichum thermophile
Applied Microbiology and Biotechnology, 2004Co-Authors: Evangelos Topakas, Haralambos Stamatis, Paul Christakopoulos, Peter BielyAbstract:A feruloyl esterase (StFAE-A) produced by Sporotrichum thermophile was purified to homogeneity. The purified homogeneous preparation of native StFAE-A exhibited a molecular mass of 57.0±1.5 kDa, with a mass of 33±1 kDa on SDS-PAGE. The pI of the enzyme was estimated by cation-exchange chromatofocusing to be at pH 3.1. The enzyme activity was optimal at pH 6.0 and 55–60 °C. The purified esterase was stable at the pH range 5.0–7.0. The enzyme retained 70% of activity after 7 h at 50 °C and lost 50% of its activity after 45 min at 55 °C and after 12 min at 60 °C. Determination of kcat/Km revealed that the enzyme hydrolyzed methyl p-coumarate 2.5- and 12-fold more efficiently than methyl caffeate and methyl ferulate, respectively. No activity on methyl sinapinate was detected. The enzyme was active on substrates containing ferulic acid ester linked to the C-5 and C-2 linkages of arabinofuranose and it hydrolyzed 4-nitrophenyl 5-O-trans-feruloyl-α-l-arabinofuranoside (NPh-5-Fe-Araf) 2-fold more efficiently than NPh-2-Fe-Araf. Ferulic acid (FA) was efficiently released from destarched wheat bran when the esterase was incubated together with xylanase from S. thermophile (a maximum of 34% total ferulic acid released after 1 h incubation). StFAE-A by itself could release FA, but at a level almost 47-fold lower than that obtained in the presence of xylanase. The potential of StFAE-A for the synthesis of various phenolic acid esters was tested using a ternary water-Organic Mixture consisting of n-hexane, 1-butanol and water as a reaction system.
Alan D. Pérez - One of the best experts on this subject based on the ideXlab platform.
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Liquid–Liquid Equilibrium and Molecular Toxicity of Active and Inert Diluents of the Organic Mixture Tri-iso-octylamine/Dodecanol/Dodecane as a Potential Liquid Membrane for Lactic Acid Removal
Journal of Chemical & Engineering Data, 2019Co-Authors: Alan D. Pérez, Verónica M. Gómez, Sneyder Rodríguez-barona, Javier FontalvoAbstract:Lactic acid can be in situ removed from a fermentation broth through reactive liquid extraction or a liquid membrane to enhance the fermentation process. The Organic Mixture tri-iso-octylamine (TiOA)/dodecanol/dodecane at 10 vol % of the amine is a potential Organic Mixture for lactic acid removal. Liquid–liquid equilibria with lactic acid aqueous solutions and molecular toxicity on the bacteria Lactobacillus casei ATCC 393 were measured with several dodecanol proportions in dodecane (0 to 90 vol %) and 10 vol % TiOA as potential solvents or membrane phases for LA removal from a fermentation broth. Effects of the Organic phase on the bacteria as cell growth, biomass production, glucose consumption, productivity, and product to biomass yield are analyzed. Dodecanol increases the lactic acid chemical equilibrium constant for the liquid–liquid equilibria, while increasing the molecular toxicity on the bacteria. However, for dodecanol concentrations from 30 to 40 vol % the value of the chemical equilibrium co...
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liquid liquid equilibrium and molecular toxicity of active and inert diluents of the Organic Mixture tri iso octylamine dodecanol dodecane as a potential liquid membrane for lactic acid removal
Journal of Chemical & Engineering Data, 2019Co-Authors: Alan D. Pérez, Verónica M. Gómez, Sneyder Rodriguezbarona, Javier FontalvoAbstract:Lactic acid can be in situ removed from a fermentation broth through reactive liquid extraction or a liquid membrane to enhance the fermentation process. The Organic Mixture tri-iso-octylamine (TiOA)/dodecanol/dodecane at 10 vol % of the amine is a potential Organic Mixture for lactic acid removal. Liquid–liquid equilibria with lactic acid aqueous solutions and molecular toxicity on the bacteria Lactobacillus casei ATCC 393 were measured with several dodecanol proportions in dodecane (0 to 90 vol %) and 10 vol % TiOA as potential solvents or membrane phases for LA removal from a fermentation broth. Effects of the Organic phase on the bacteria as cell growth, biomass production, glucose consumption, productivity, and product to biomass yield are analyzed. Dodecanol increases the lactic acid chemical equilibrium constant for the liquid–liquid equilibria, while increasing the molecular toxicity on the bacteria. However, for dodecanol concentrations from 30 to 40 vol % the value of the chemical equilibrium co...