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Synthesis of Functional Lipid from Glyceryl Monooleate and Conjugated Linoleic Acid by Enzymatic Reaction

Glyceryl Monooleate와 Conjugated Linoleic Acid로부터 효소적 반응을 이용한 기능성 유지 합성

  • Jeon, Mi-Sun (Dept. of Food Science and Technology, Chungnam National University) ;
  • Lee, Ki-Teak (Dept. of Food Science and Technology, Chungnam National University)
  • Published : 2009.08.31

Abstract

Diacylglycerol (DAG) were synthesized by enzymatic esterification of glyceryl monooleate (GMO) and conjugated linoleic acid (CLA) in a shaking water bath. The reaction was catalyzed by Lipozyme TLIM (immobilized lipase from Thermomyces lanuginosa). Effects of reaction time, molar ratio, enzyme road and molecular sieves were studied. Results of normal-phase high performance liquid chromatography (NP-HPLC) analysis were performed. At 1:1, 2:1 and 3:1 (GMO : CLA) molar ratio and Lipozyme TLIM of 20% amount, DAG were produced in 42.6, 54.4 and 54.6 area% in 1 hr, respectively. When different Lipozyme TLIM amounts (2, 5, 10, 20%) were used with 2:1 (GMO : CLA) molar ratio, DAG were produced 21.4 (24 hr), 51.7 (12 hr), 56.2 (6 hr) and 54.4 (1 hr) area%, respectively. The reaction in the absence of molecular sieves increased DAG contents. The maximum DAG concentration conditions were obtained with molar ratio of 2:1 (GMO : CLA), lipase concentration of 10% (of substrate), 10% molecular sieves and reaction time of 6 hours at 55$^{\circ}C$. Under this reaction condition, produced DAG-rich oil was composed of 69 area% DAG, 7.9 area% TAG, 2 area% FFA, and 21.1 area% MAG.

DAG 합성율이 가장 높은 몰비율을 선정하기 위하여 GMO와 CLA의 몰비율을 1:1, 2:1 그리고 3:1로 하고 Lipozyme TLIM은 기질 질량의 20%를 사용하여 반응하였다. 그 결과 총 DAG의 함량은 1 hr일 때 높게 나타나 몰비율이 1:1, 2:1, 3:1(GMO:CLA)일 경우 각각 42.6, 54.4 그리고 54.6 area%로 나타났고, DAG 함량이 높은 2:1과 3:1의 기질몰비율 조건 중 MAG와 TAG의 함량이 낮았던 2:1 비율을 가장 적합한 비율로 선정하였다. 적합한 몰비율로 선정된 GMO와 CLA의 몰비율 2:1에 효소량을 2%, 5%, 10% 그리고 20%를 사용하여 효소량에 따른 DAG 합성율을 살펴 본 결과, 효소량을 10% 사용하였을 경우, 반응 6 hr에서 56.2 area%의 가장 높은 DAG 함량을 나타내었다. 이어 Lipozyme TLIM을 10% 사용하여 GMO와 CLA의 몰비율 2:1에 molecular sieves를 전체 기질 질량의 10% 사용하여 molecular sieves 첨가의 유, 무에 따른 DAG 합성율을 알아보았다. 그 결과 효소량을 5%를 사용하여 반응하였을 경우 molecular sieves를 첨가하지 않았을 때보다 총 DAG 함량이 약 30%의 증가율을 보였고, 효소량을 10% 사용하였을 경우 23%의 증가율을 보였다. 이상에서 DAG가 가장 많이 합성된 반응조건은 몰비율이 GMO와 CLA가 2:1이고, Lipozyme TLIM과 molecular sieves의 양이 각각 기질 질량의 10%씩 사용되었을 때이었다. 이 반응 조건으로 6 hr 반응하였을 때 최대의 DAG량이 생성되어 그 함량은 69 area%로 나타났으며, 1,3-DAG과 1,2-DAG의 지방산 조성변화를 살펴 본 결과 1,2-DAG의 지방산 조성 중 CLA의 함량은 반응 시간이 지날수록 증가한 반면, 1,3-DAG의 CLA 함량은 1 hr에서 24 hr까지 증가하였다가 72 hr에서 감소함을 보였다.

Keywords

References

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