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ammonium sulphate system
S500
Abstracts / Journal of Biotechnology 136S (2008) S496–S505
affinity for water. To date, there is no single method that could be simple and efficient for the separation of 2,3-BD (Xiu and Zeng, 2008). In this paper, a novel aqueous two-phase system (ATPS) of isopropanol/ammonium sulfate was used for the recovery of 2,3-BD. The influences of isopropanol and ammonium sulfate on the partitioning behavior were studied with filtrated broth. It has been shown that the increase in partitioning coefficients and recoveries of 2,3-BD and acetoin with increasing concentrations of isopropanol and ammonium sulfate, and the partition coefficients of glucose decreased obviously with a low value. At the ammonium sulfate concentration of 20% (w/w) and isopropanol concentration of 34% (w/w), the partition coefficients of 2,3-BD, acetoin and glucose reached 8.29, 8.98 and 0.030, respectively; the recoveries of 2,3-BD and acetoin were 91.36% and 91.62%, respectively. In addition, aqueous two-phase extractions (ATPE) were performed both in 10 and 3 kg scales by using fermentation broths without filtration. During three batches of 3 kg scale ATPE, the removal ratios of cells were all above 99%, and the concentrations of soluble protein in the top phase decreased to about 0.05 g L−1 . The cells in ATPS were separated and re-used as seeds for subsequent fermentation. The results show that the cells recycle fermentation is practical, which eliminates the inoculum preparation, and saves more time and cost. The results above mentioned demonstrated the feasibility of ATPE for recovery of 2,3-BD by isopropanol/ammonium sulfate system, which has potential uses in industrialization.
References Afschar, A.S., Vaz Rossell, C.E., Jonas, R., Chanto, A.Q., Schaller, K., 1993. Microbial production and downstream processing of 2,3-butanediol. J. Biotechnol. 27, 317–329. Xiu, Z.L., Zeng, A.P., 2008. Present state and perspective of downstream processing of biologically produced 1,3-propanediol and 2,3-butanediol. Appl. Microbiol. Biotechnol. 78, 917–926.
doi:10.1016/j.jbiotec.2008.07.1171 V6-P-009 Microwave-assisted aqueous two-phase extraction of piceid, resveratrol and emodin from Polygonum cuspidatum by ethanol/ammonium sulphate system Hui Wang, Yuesheng Dong, Zhilong Xiu ∗ Department of Bioscience and Biotechnology, School of Environmental and Biological Science and Technology, Dalian University of Technology, Dalian, PR China E-mail address: [email protected] (Z. Xiu). Piceid, resveratrol, and emodin, as the main components of a traditional Chinese herbal medicine Polygonum cuspidatum, are widely used in medicine, health products and cosmetic industries. Traditional extraction needs large amount of ethanol and long time. Moreover, filtration and concentration are needed before purification by column chromatography (Xiang et al., 2005), which results in the low yield and high cost of the products. Microwave-assisted extraction (MAE) has been proved to be an alternative method for the extraction of active components from herbs due to its advantages such as short time, less solvent and high efficiency (Pan et al., 2000), and short-chain alcohol/salt system has been used as a novel aqueous two-phase system to purify natural compounds because of its advantages such as low cost and easy recovery of alcohol by evaporation (Tan et al., 2002).
In this paper, microwave-assisted aqueous two-phase extraction of effective constituents including piceid, resveratrol and emodin directly from Polygonum cuspidatum by ethanol/ammonium sulphate system was performed and the partition behavior of them was investigated. The results indicated that partition coefficient and yield increase with the increasing of ethanol and ammonium sulphate. Based on the results, phase composition on phase ratio and yield of piceid, resveratrol and emodin was studied and the system consisting of 25% (w/w) ethanol and 21% (w/w) ammonium sulphate was selected as the optimum system for the extraction to get equal yield of piceid, and that of resveratrol and emodin 1.1 and 1.9 times higher, respectively, than that by microwave-assisted extraction and heat reflux extraction. Microwave-assisted aqueous two-phase extraction integrates extraction, clarification and concentration into a single step and greatly reduce filtration load to get higher yield at lower cost. It possesses the advantages of both microwave-assisted extraction and aqueous two-phase extraction, so it is a potential method for the extraction and purification of target products.
References Pan, X.J., Liu, H.Z., Jia, G.H., Shu, Y.Y., 2000. Microwave-assisted extraction of glycyrrhizic acid from licorice root. Biochem. Eng. J. 5, 173–177. Tan, T.W., Huo, Q., Ling, Q., 2002. Purification of glycyrrhizin from Glycyrrhiza uralensis Fisch with ethanol/phosphate aqueous two phase system. Biotechnol. Lett. 24, 1417–1420. Xiang, H.Y., Zhou, C.S., Lei, Q.F., Chen, L.S., 2005. Studies on separation and purification of piceid from Polygonum cuspidatum by macroporous adsorption resin (in Chinese). Chin. Pharm. J. 40, 96–98.
doi:10.1016/j.jbiotec.2008.07.1172 V6-P-010 Separation of 2,3-butanediol from fermentation broths with ethanol/phosphate aqueous two-phase system Bo Jiang, Zhigang Li, Daijia Zhang, Zhilong Xiu ∗ Department of Bioscience and Biotechnology, School of Environmental and Biological Science and Technology, Dalian University of Technology, Dalian, PR China E-mail address: [email protected] (Z. Xiu). 2,3-Butanediol is an important promising chemical which can be used as a variety of chemical feedstocks and liquid resources (Syu, 2001). The high boiling point and high affinity for water make it difficult for 2,3-butanediol to be separated from complicated fermentation broths. Many methods have been investigated for the recovery of 2,3-butanediol including steam stripping, liquid–liquid extraction, and solvent extraction, but none of them has proved to be simple and efficient (Xiu and Zeng, 2008). Aqueous two-phase extraction has been widely applied in separation of biomacromlecules, such as proteins and nucleic acids. Short-chain alcohols or hydrophilic organic solvents and salts can also be able to form aqueous two-phase systems which has some advantages over traditional ones such as low cost of materials and easy recovery of products (Tan et al., 2002). A hydrophilic organic solvent/salt aqueous two-phase system was formed by ethanol and dipotassium hydrogen phosphate. Influencing factors including phase diagram and phase composition on partition of 2,3-butanediol, removal of cells and biomacromolecules were investigated. As indicated by the results, 2,3-butanediol can be extracted efficiently to the top phase. The partition coefficient and recovery of 2,3-butanediol reached up to 27.75% and 99.1%, respectively, and the selective coefficient of 2,3-
Abstracts / Journal of Biotechnology 136S (2008) S496–S505
affinity for water. To date, there is no single method that could be simple and efficient for the separation of 2,3-BD (Xiu and Zeng, 2008). In this paper, a novel aqueous two-phase system (ATPS) of isopropanol/ammonium sulfate was used for the recovery of 2,3-BD. The influences of isopropanol and ammonium sulfate on the partitioning behavior were studied with filtrated broth. It has been shown that the increase in partitioning coefficients and recoveries of 2,3-BD and acetoin with increasing concentrations of isopropanol and ammonium sulfate, and the partition coefficients of glucose decreased obviously with a low value. At the ammonium sulfate concentration of 20% (w/w) and isopropanol concentration of 34% (w/w), the partition coefficients of 2,3-BD, acetoin and glucose reached 8.29, 8.98 and 0.030, respectively; the recoveries of 2,3-BD and acetoin were 91.36% and 91.62%, respectively. In addition, aqueous two-phase extractions (ATPE) were performed both in 10 and 3 kg scales by using fermentation broths without filtration. During three batches of 3 kg scale ATPE, the removal ratios of cells were all above 99%, and the concentrations of soluble protein in the top phase decreased to about 0.05 g L−1 . The cells in ATPS were separated and re-used as seeds for subsequent fermentation. The results show that the cells recycle fermentation is practical, which eliminates the inoculum preparation, and saves more time and cost. The results above mentioned demonstrated the feasibility of ATPE for recovery of 2,3-BD by isopropanol/ammonium sulfate system, which has potential uses in industrialization.
References Afschar, A.S., Vaz Rossell, C.E., Jonas, R., Chanto, A.Q., Schaller, K., 1993. Microbial production and downstream processing of 2,3-butanediol. J. Biotechnol. 27, 317–329. Xiu, Z.L., Zeng, A.P., 2008. Present state and perspective of downstream processing of biologically produced 1,3-propanediol and 2,3-butanediol. Appl. Microbiol. Biotechnol. 78, 917–926.
doi:10.1016/j.jbiotec.2008.07.1171 V6-P-009 Microwave-assisted aqueous two-phase extraction of piceid, resveratrol and emodin from Polygonum cuspidatum by ethanol/ammonium sulphate system Hui Wang, Yuesheng Dong, Zhilong Xiu ∗ Department of Bioscience and Biotechnology, School of Environmental and Biological Science and Technology, Dalian University of Technology, Dalian, PR China E-mail address: [email protected] (Z. Xiu). Piceid, resveratrol, and emodin, as the main components of a traditional Chinese herbal medicine Polygonum cuspidatum, are widely used in medicine, health products and cosmetic industries. Traditional extraction needs large amount of ethanol and long time. Moreover, filtration and concentration are needed before purification by column chromatography (Xiang et al., 2005), which results in the low yield and high cost of the products. Microwave-assisted extraction (MAE) has been proved to be an alternative method for the extraction of active components from herbs due to its advantages such as short time, less solvent and high efficiency (Pan et al., 2000), and short-chain alcohol/salt system has been used as a novel aqueous two-phase system to purify natural compounds because of its advantages such as low cost and easy recovery of alcohol by evaporation (Tan et al., 2002).
In this paper, microwave-assisted aqueous two-phase extraction of effective constituents including piceid, resveratrol and emodin directly from Polygonum cuspidatum by ethanol/ammonium sulphate system was performed and the partition behavior of them was investigated. The results indicated that partition coefficient and yield increase with the increasing of ethanol and ammonium sulphate. Based on the results, phase composition on phase ratio and yield of piceid, resveratrol and emodin was studied and the system consisting of 25% (w/w) ethanol and 21% (w/w) ammonium sulphate was selected as the optimum system for the extraction to get equal yield of piceid, and that of resveratrol and emodin 1.1 and 1.9 times higher, respectively, than that by microwave-assisted extraction and heat reflux extraction. Microwave-assisted aqueous two-phase extraction integrates extraction, clarification and concentration into a single step and greatly reduce filtration load to get higher yield at lower cost. It possesses the advantages of both microwave-assisted extraction and aqueous two-phase extraction, so it is a potential method for the extraction and purification of target products.
References Pan, X.J., Liu, H.Z., Jia, G.H., Shu, Y.Y., 2000. Microwave-assisted extraction of glycyrrhizic acid from licorice root. Biochem. Eng. J. 5, 173–177. Tan, T.W., Huo, Q., Ling, Q., 2002. Purification of glycyrrhizin from Glycyrrhiza uralensis Fisch with ethanol/phosphate aqueous two phase system. Biotechnol. Lett. 24, 1417–1420. Xiang, H.Y., Zhou, C.S., Lei, Q.F., Chen, L.S., 2005. Studies on separation and purification of piceid from Polygonum cuspidatum by macroporous adsorption resin (in Chinese). Chin. Pharm. J. 40, 96–98.
doi:10.1016/j.jbiotec.2008.07.1172 V6-P-010 Separation of 2,3-butanediol from fermentation broths with ethanol/phosphate aqueous two-phase system Bo Jiang, Zhigang Li, Daijia Zhang, Zhilong Xiu ∗ Department of Bioscience and Biotechnology, School of Environmental and Biological Science and Technology, Dalian University of Technology, Dalian, PR China E-mail address: [email protected] (Z. Xiu). 2,3-Butanediol is an important promising chemical which can be used as a variety of chemical feedstocks and liquid resources (Syu, 2001). The high boiling point and high affinity for water make it difficult for 2,3-butanediol to be separated from complicated fermentation broths. Many methods have been investigated for the recovery of 2,3-butanediol including steam stripping, liquid–liquid extraction, and solvent extraction, but none of them has proved to be simple and efficient (Xiu and Zeng, 2008). Aqueous two-phase extraction has been widely applied in separation of biomacromlecules, such as proteins and nucleic acids. Short-chain alcohols or hydrophilic organic solvents and salts can also be able to form aqueous two-phase systems which has some advantages over traditional ones such as low cost of materials and easy recovery of products (Tan et al., 2002). A hydrophilic organic solvent/salt aqueous two-phase system was formed by ethanol and dipotassium hydrogen phosphate. Influencing factors including phase diagram and phase composition on partition of 2,3-butanediol, removal of cells and biomacromolecules were investigated. As indicated by the results, 2,3-butanediol can be extracted efficiently to the top phase. The partition coefficient and recovery of 2,3-butanediol reached up to 27.75% and 99.1%, respectively, and the selective coefficient of 2,3-