lipase catalyzed sugar esters synthesis in ionic liquids

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Disclaimer 저작자표시. 귀하는 원저작자를 표시하여야 합니다. 동일조건변경허락. 귀하가 이 저작물을 개작, 변형 또는 가공했을 경우 에는, 이 저작물과 동일한 이용허락조건하에서만 배포할 수 있습니다. 工學碩士學位請求論文 리파아제 촉매를 이용한 이온성 액체에서의 당 에스터 합성 LIPASE-CATALYZED SUGAR ESTERS SYNTHESIS IN IONIC LIQUIDS 2009 년 2 월 仁荷大學校 大學院 生物工學科 NGUYEN MINH HIEP 工學碩士學位請求論文 리파아제 촉매를 이용한 이온성 액체에서의 당 에스터 합성 LIPASE-CATALYZED SUGAR ESTERS SYNTHESIS IN IONIC LIQUIDS 2009 년 2 월 指導敎授 具 潤 謨 이 論文을 工學碩士學位 論文으로 提出함 仁荷大學校 大學院 生物工學科 NGUYEN MINH HIEP LIPASE-CATALYZED SUGAR ESTERS SYNTHESIS IN IONIC LIQUIDS By NGUYEN MINH HIEP A THESIS submitted to the faculty of INHA UNIVERSITY In partial fulfillment of the requirements For the degree of MASTER OF SCIENCE Department of Marine Science and Biological Engineering February 2009 TRƯỜNG ĐẠI HỌC KỸ THUẬT INHA Luận văn thạc sĩ NGUYỄN MINH HIỆP TỔNG HỢP SUGAR ESTERS TRONG IONIC LIQUIDS DƯỚI SỰ XÚC TÁC CỦA LIPASE Cán bộ hướng dẫn GS. TS. YOON-MO KOO Khoa Hải Dương Học và Công Nghệ Sinh Học Tháng 2 năm 2009 이 論文을 NGUYEN MINH HIEP 의 碩士學位 論文으로 認定함 2009 年 2 月 主審 副審 委員 LIPASE-CATALYZED SUGAR ESTERS SYNTHESIS IN IONIC LIQUIDS By NGUYEN MINH HIEP Supervised by Dr. YOON-MO KOO Department of Marine Science and Biological Engineering February 2009 i ACKNOWLEDGEMENT In the first place, I would like to express my gratitude to Professor Koo Yoon-Mo, who has given me the opportunity to live and work in Korea. I will never forget the useful lessons he has taught me and I am grateful for this. I also extend my thanks to all professors in Department of Marine Science and Biological Engineering, Inha University for their lectures together with their cherishing and support. I really appreciate Doctor Ha Sung Ho and Doctor Lee Sang Hyun for their help and guidance. I am grateful to all of the members in Bionanoprocess laboratory, who are not only friendly but also enthusiastic to help me during the time I lived and studied in Korea. These will become the nice memories about the period I was in Korea. I wish to thank ERC organization for supporting me in research and trips to the conference at Cheju island of Korea and Dalian of China. Warm thanks also go to all of the Vietnamese and Korean friends at Inha University who have shared and helped me in my living and work. Finally, I would like to express my biggest thank to my family, my partner – Nguyen Thi Huynh Nga as well as her family for their constant help, availability and motivation, thus allowing me to be more optical and confident when things were not going on so well. ii ABSTRACT Fatty acid sugar esters known as bio-sufactants can be produced from renewable resources. They are tasteless, odorless, nontoxic, non-irritant, non-ionic and cover a wide range of hydrophilic-lipophilic balance which explain well their increasing importance in numerous areas such as food, cosmetic, detergent and pharmaceutical industry. While most of these compounds are chemically synthesized, the enzymatic synthesis of sugar esters has garnered considerable interest by reasons of milder and simpler conditions, higher selectivity, and greener process. Nowadays, ionic liquids (ILs) are known as ‘green’ alternatives to volatile organic solvents (VOCs) in electrochemical, synthetic, separation and chemical processes because of their characteristics such as non-volatility, non- flammability, solvating power for dissolving a wide range of organic, inorganic and polymeric materials and their excellent chemical and thermal stability. In addition, ILs have been described as ‘designer solvents’ since their physicochemical properties such as polarity, hydrophobicity, density and viscosity can be adjusted to suit the requirements for a particular process by altering the anions, and (or) cations. Besides, ultrasound irradiation is known as a useful tool for strengthening mass transfer of liquid-liquid heterogeneous systems. In this study, the synthesis of sugars esters by using lipase-catalyzed esterification with fatty acids in ionic liquids mixtures by using supersaturated sugar solution with the combination of ultrasound irradiation was investigated. Firstly, the lipase-catalyzed synthesis of glucose fatty acid esters using ionic liquid mixtures was investigated. Although the activity of Novozym 435 in a 1-butyl-3- methylimidazolium trifluoromethanesulfonate ([Bmim][TfO]) and 1-butyl-3- methylimidazolium bis[(trifluoromethyl)sulfonyl]amide ([Bmim][Tf2N]) mixture (1:1 v/v) was somewhat lower than that in pure [Bmim][TfO] which showed the highest enzyme iii activity, the stability of Novozym 435 was significantly increased. Specifically, the activity of Novozym435 was increased from 1.1 to 2.9 mmol min -1 g -1 by using supersaturated glucose solution in this mixture, compared with reaction using saturated solution. After 5 times reuse of Novozym 435, 86% of initial activity was remained in this mixture, while the residual activity in pure [Bmim][TfO] was 36%. Therefore, the productivity obtained by using IL mixtures was higher than those in pure ILs. Secondly, the application of ultrasound irradiation in enhancing lipase activity in the synthesis of sugar ester using ionic liquids was investigated. In the lipase-catalyzed esterifications of glucose with vinyl laurate or lauric acid in [Bmim][TfO], 2.4 and 4.7 times higher activities, respectively, were obtained by using supersaturated solution under ultrasound irradiation than those in conventional method. In the lipase-catalyzed transesterification using [Bmim][PF 6 ], enzyme activity was 5.8 times increased by ultrasound irradiation. Finally, the optimization of the synthesis of glucose fatty acid ester in IL mixtures was investigated by using supersaturated glucose solution. The effect of IL mixtures ratio, substrate ratio, water content, lipase content, and temperature on the activity and stability of lipase was also studied. The highest yield of sugar ester was obtained in a [Bmim][TfO] and [Omim][Tf 2 N] mixture (9:1 v/v), while Novozym 435 showed the optimal stability and activity in a mixture of [Bmim][TfO]:[Omim][Tf 2 N] (1:1, v/v). Reuse of lipase and ILs was successfully carried out with optimized reaction conditions. After 5 times reuse of Novozym 435 and ILs, 78% of initial activity was remained. [...]... supersaturated sugar solution prepared by water- mediated method under effects of ultrasound irradiation was investigated This investigation includes three parts: 1 Lipase- catalyzed synthesis of glucose fatty acid ester using ionic liquid mixtures 2 Ultrasound-enhanced lipase activity in the synthesis of sugar ester using ionic liquids 3 Optimization of lipase- catalyzed glucose esters synthesis in ionic liquids. .. stimulation Lin et al (1997) reported that the activity of inulinase could achieve a 60% rise in the presence of ultrasound of 20W [52] Zong et al (2000) observed a more significant increase in activity in the presence of ultrasound for immobilized enzymes than for free enzymes [53] 10 2 OBJECTIVES In this thesis, enzymatic synthesis of sugar esters by using lipase- catalyzed reaction in ILs with supersaturated... in IL and IL mixtures 19 4.1.2 Time courses for the lipase- catalyzed esterification of glucose with vinyl laurate in pure ILs and IL mixtures 24 4.1.3 Residual activity of Novozym 435 after reuse in pure ILs and IL mixtures 25 4.2 Ultrasound-enhanced lipase activity in the synthesis of sugar ester using ionic liquids 27 4.2.1 Dissolution rate of glucose in. .. the substrate and product, thereby, decreasing the substrate and product inhibition observed in water, and hence, increasing the catalyst activity and productivity [28] The feasibility of using isolated enzyme in IL media has also been demonstrated Erbeldinger et al reported the thermolysin-catalysed synthesis of Zaspartame in [Bmim][PF6]/H2O (95/5, v/v) [29] In addition, the potential of ILs used as... hexafluorophosphate xi 1 INTRODUCTION 1.1 Fatty acid sugar esters Fatty acid sugar esters, well known as bio-sufactants are produced from renewable resources [1] They are tasteless, odorless, nontoxic and non-irritating, non -ionic and their high biodegradability explains their increasing importance in numerous areas such as pharmaceuticals, cosmetics, detergents, and the food industry [2, 3] Their physicochemical... solvents in biocatalytic processes could potentially overcome these problems and open up some exciting new opportunities Ionic liquids with their unique properties are receiving increasing attention as solvents for organic synthesis in general and catalytic processes in particular Depending on their structure, they can be immiscible with water or e g alkanes, which render them useful for performing catalytic... various mono and disaccharides in ILs containing [Cl]- [40] However, ILs containing [Cl]- cannot be used to carry out enzyme reactions due to the inactivation of most enzymes [41] Only ILs containing the dicyanamide ([dca]) anion have been reported to be good 7 solvents for sugar dissolution and enzyme reactions, but Candida Antarctica lipase B irreversibly deactivated in [Bmim][dca] after reaction... [Bmim][PF6] were used as reaction media in the lipase- catalyzed transesterification of glucose with a fatty acid vinyl ester However, the solubility of glucose in these ILs is very low [37] To overcome this problem, supersaturated sugar solutions in ILs were recently used to carry out enzymatic synthesis of sugar esters [44] The supersaturated solution, a solution which contains more dissolved solute than the... having a melting point below 100oC, are composed entirely of ions and possess a wide liquid temperature range, in some case in excess of 400oC [12] If the salt’s melting point is below room temperature (~ 25oC), the IL is called a room-temperature ionic liquid (RTIL) The term IL covers inorganic as well as organic molten salts [13] The first [EtNH3][NO3] was reported in 1914, but the most popular ionic. .. length of the alkyl chain in the cation and/or shape or symmetry [18, 19] Some typical cation/anion combinations comprising the main families of ILs are illustrated in Figure 1 Figure 1 Some typical cation/anion combinations in ILs Recently, ionic liquids have been extensively evaluated as environmentally friendly or “green” alternatives to conventional volatile organic solvents (VOCs) in many applications . SUGAR ESTERS SYNTHESIS IN IONIC LIQUIDS 2009 년 2 월 指導敎授 具 潤 謨 이 論文을 工學碩士學位 論文으로 提出함 仁荷大學校 大學院 生物工學科 NGUYEN MINH HIEP LIPASE-CATALYZED SUGAR ESTERS SYNTHESIS IN IONIC LIQUIDS. irradiation in enhancing lipase activity in the synthesis of sugar ester using ionic liquids was investigated. In the lipase-catalyzed esterifications of glucose with vinyl laurate or lauric acid in. sugars esters by using lipase-catalyzed esterification with fatty acids in ionic liquids mixtures by using supersaturated sugar solution with the combination of ultrasound irradiation was investigated.