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

Abstract

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 degrees C, respectively. Substrate molar ratio was varied from 1:1 to 1:4. The highest incorporation of oleic acid (37.7%) was at 45 degrees 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 degrees 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 degrees C. Lipozyme RM IM gave highest oleic acid incorporation (49.4%) at 65 degrees 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.

Production of human milk fat substitute with medium-chain fatty acids by lipase-catalyzed acidolysis: Optimization by response surface methodology

“…The TAG composition of the products in lipase-catalyzed reaction was reported to depend on the substrate molar ratio (Srivastava et al, 2006;Yang et al, 2003). In our study, the incorporation of caprylic and capric acid was found to increase with increasing substrate molar ratio, whereas the incorporation of palmitic acid decreased with increasing substrate molar ratio, which may be attributed to the ratio of reaction substrates.…”

“…Vegetable oils (palm, soybean, hazelnut and amaranth oil), lard, butter and tripalmitin have been used as substrate to produce human milk fat substitutes. Substrate molar ratio, enzyme amount, reaction time and temperature effects on the composition of human milk fat substitutes produced have been studied (Jimenez et al, 2010;Karabulut, Turan, Vural, & Kayahan, 2007;Li et al, 2010;Nielsen, Yang, Xu, & Jacobsen, 2006;Ozcelik, Karaali, Sahin Yesilcubuk, & Kahveci, 2008;Pina-Rodriguez & Akoh, 2009;Sahin et al, 2005;Srivastava, Akoh, Chang, Lee, & Shaw, 2006;Sørensen et al, 2009;Yang et al, 2003).…”

An ultraviolet spectrophotometric assay for the screening of sn-2-specific lipases using 1,3-O-dioleoyl-2-O-α-eleostearoyl-sn-glycerol as substrate

“…Intensive research is being carried out to make use of the specifi cities and preferences of some lipases for synthesizing lipids with well-defi ned FA compositions and patterns of distribution, which could be used in the production of human dietary supplements and edible oils ( 26,27 ).…”

Enzymatic Preparation of Human Milk Fat Substitutes and Their Oxidation Stability

“…To date, several methods have been used to prepare HMFSs. In particular, enzymatic acidolysis has been commonly used in several studies for HMFS preparation 5,6 .…”