JOURNAL OF OLEO SCIENCE Copyright 2005 by Japan Oil Chemists Society JOS Setsuko HARA, Ayato KANDA and Yoichiro TOTANI Faculty of Engineering, Seikei University 3-1 Kichijoji-kitamachi 3, Musashino-shi, Tokyo 180-8633, JAPAN Edited by K. Miyashita, Hokkaido Univ., and accepted April 18, 2005 (received for review March 7, 2005) Abstract: Structured lipids with n-6/n-3 polyunsaturated fatty acids PUFA or medium chain fatty acids MC at optimal ratios were prepared by lipase catalyzed transacylation of tuna orbital oil consisting of about 40 n-3 PUFA such as eicosapentaenoic acid EPA and docosahexaenoic acid DHA with purified linoleic acid or MC, such as caprylic, capric, and lauric acids, at 60. 1, 3- Specific immobilized lipases of Lipozyme TL-IM and Lipozyme RM- IM originated from Thermomyces lanuginosus and Rhizomucor miehei were used in the preparation, respectively. By controlling enzyme amount from 20-80 IUN for Lipozyme TL-IM and 1-12 BAUN for Lipozyme RM-IM with reaction time from 4 to 24h, structured lipids at n-6/n-3 ratios from 1.3 till 6.9 were easily prepared by the reaction of tuna orbital oil with highly purified linoleic acid. MC was introduced into tuna orbital oil at various MC/n-3 PUFA ratios such as 0.67-1.89 for caprylic, 0.73-2.85 for capric, and 0.91-4.08 for lauric acids, respectively during transacylation for 32 h. Lipozyme RM-IM was clearly shown a better enzyme for the present transacylation, compared to Lipozyme TL-IM in consideration of the above ratio values and triglycerides recovery. Key words: transacylation, Lipozyme TL-IM, Lipozyme RM-IM, n-3 PUFA, n-6 PUFA, MC 1 3 6 1 1999 6 3 4 6 3 1 6 3 2 3 Correspondence to: Setsuko HARA, Faculty of Engineering, Seikei University, 3-1 Kichijoji-kitamachi 3, Musashino-shi, Tokyo 180-8633, JAPAN E-mail: shara@ch.seikei.ac.jp Journal of Oleo Science ISSN 1345-8957 print / ISSN 1347-3352 online http://jos.jstage.jst.go.jp/en/ 473
S. Hara, A. Kanda and Y. Totani 2 2 2 2 2 4 b 5 6 2 1 3 6 7 8 1 3 2 8 8 1 3 2 8 2 1 3 6 Riozopus delemar 1 3 65 8 1 2 8 3 8 Candida rugosa 9 10 13 Fig. 1 3 6 6 3 2 1 3 3 2 2 1 2 1 1 Fig. 1 Preparation of Structured Lipids. 474
2 1 2 6 3 3 8 10 12 Table 1 2 1 3 1 3 Thermomyces lanuginosus 75 Rhizomucor miehei 5 6 2 2 1 4 1 2 2 2 2 1 60 63 212 m 400 130 4 6 72 30 85 15 800 Table 1 3 2 1497 2 3 2 3 1 14 17 10 0 25 25 50 3 10 170 2 174 1 186 2 200 5 250 2 3 2 5 10 9 1 2 3 3 1 4 6 250 210 295 325 2 99 2 0 8 0 7 2 3 4 15 Table 1 Fatty Acid Composition of Substrates. Tuna Oil Linoleic Acid Caprylic Acid Capric Acid Lauric Acid Caprylic Acid 99.5 Capric Acid 99.3 Lauric Acid 99.1 Linoleic Acid (n-6 PUFA) 1.3 95.2 EPA + DHA (n-3 PUFA) 39.4 Others 59.3 4.8 0.5 0.7 0.9 475
S. Hara, A. Kanda and Y. Totani 2 4 2 4 1 Fig. 2 6 60 2 1 1 3 1 5 100 2 4 2 3 3 2 4 1 8 10 12 6 60 3 3 1 8 Fig. 3 6 1 3 80 47 9 1 4 8 6 44 6 6 60 80 3 3 20 1 3 40 3 20 1 3 20 2 6 3 6 3 Table 2 80 8 24 Table 2 6 3 59 65 1 63 68 6 3 1 3 3 2 1 7 2 5 Table 2 24 63 69 2 6 3 2 8 6 9 2 3 4 1 6 3 Fig. 2 Procedure of Transacylation. Fig. 3 Effects of Enzyme Amounts on PUFA-TG Preparation Catalyzed by Lipozyme TL-IM or Lipozyme RM-IM. 476
Table 2 Comparison of n-6/n-3 Ratio of PUFA-TG Prepared. Lipozyme TL-IM Lipozyme RM-IM Effect of Enzyme Amounts Enzyme Amounts (Unit) 20 IUN 40 IUN 60 IUN 80 IUN 1 BAUN 4 BAUN 8 BAUN 12 BAUN Reaction Time (h) 8 8 PUFA Content (%) 61.7 64.6 58.6 62.7 53.1 66.0 67.7 62.8 n-6/n-3 Ratio 1.3 1.5 2.3 3.2 1.9 1.7 2.2 2.5 Enzyme Amounts 80 IUN 8 BAUN Effect of Reaction Time Reaction Time (h) 4 h 8 h 16 h 24 h 4 h 8 h 16 h 24 h PUFA Content (%) 65.6 64.9 63.4 64.6 64.6 64.9 66.4 69.2 n-6/n-3 Ratio 2.8 3.3 5.2 6.9 2.3 2.6 3.3 4.1 IUN: Interesterification Unit, BAUN: Batch Acidolysis Unit 3 2 3 8 10 12 6 3 1 8 Fig. 4 6 Fig. 4 8 4 21 7 32 38 5 3 0 67 1 89 32 50 Fig. 5 10 32 48 7 3 0 73 2 85 60 12 Fig. 6 32 58 3 8 10 3 0 91 4 08 12 4 25 40 Fig. 4 Fatty Acid Composition of n-3 PUFA-C8-TG Prepared. 3 3 3 1 2 Fig. 7 4 8 80 12 24 8 10 12 32 12 10 8 15 Fig. 5 Fatty Acid Composition of n-3pufa-c10-tg Prepared. 477
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