Candida rugosa Lipase LIP1-Catalyzed Transesterification To Produce Human Milk Fat Substitute

• Published in: Journal of agricultural and food chemistry 2006-07, Vol.54 (14), p.5175-5181
• Main Authors: Srivastava, Anita, Akoh, Casimir C, Chang, Shu-Wei, Lee, Guan-Chiun, Shaw, Jei-Fu
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Candida - enzymology; Candida rugosa; enzyme activity; Esterification; Fat industries; Fats - analysis; Fatty Acids - analysis; Food industries; Fundamental and applied biological sciences. Psychology; human milk fat substitute; Kinetics; Lipase - metabolism; methyl oleate; Milk - chemistry; Milk and cheese industries. Ice creams; oleic acid; Oleic Acid - metabolism; palmitic acid; Pancreas - enzymology; structured lipids; Temperature; transesterification; triacylglycerol lipase; Triglycerides - metabolism; tripalmitin
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/jf060623h

Structured lipids (SLs) containing palmitic and oleic acids were synthesized by transesterification of tripalmitin with either oleic acid or methyl oleate as acyl donor. This SL with palmitic acid at the sn-2 position and oleic acid at sn-1,3 positions is similar in structure to human milk fat triacylglycerol. LIP1, an isoform of Candida rugosa lipase (CRL), was used as biocatalyst. The effects of reaction temperature, substrate molar ratio, and time on incorporation of oleic acid were investigated. Reaction time and temperature were set at 6, 12, and 24 h, and 35, 45, and 55 °C, respectively. Substrate molar ratio was varied from 1:1 to 1:4. The highest incorporation of oleic acid (37.7%) was at 45 °C with methyl oleate as acyl donor. Oleic acid resulted in slightly lesser (26.3%) incorporation. Generally, higher percentage incorporation of oleic acid was observed with methyl oleate (transesterification) than with oleic acid (acidolysis). In both cases percentage incorporation increased with reaction time. Incorporation decreased with increase in temperature above 45 °C. Initially, oleic acid incorporation increased with increase in substrate molar ratio up to 1:3. LIP1 was also compared with Lipozyme RM IM as biocatalysts. The tested reaction parameters were selected on the basis of maximum incorporation of C18:1 obtained during optimization of LIP1 reaction conditions. Reaction temperature was maintained at 45, 55, and 65 °C. Lipozyme RM IM gave highest oleic acid incorporation (49.4%) at 65 °C with methyl oleate as acyl donor. Statistically significant (P < 0.05) differences were observed for both enzymes. SL prepared using Lipozyme RM IM may be more suitable for possible use in human milk fat substitutes. Keywords: Acidolysis; Candida rugosa lipase; human milk fat substitute; Lipozyme RM IM; LIP1; structured lipids; transesterification; tripalmitin