IM Lipase Research Articles

Enzymatic modification of triolein: Incorporation of caproic and butyric acids to produce reduced‐calorie structured lipids

AbstractLipase‐catalyzed acidolysis of triolein with caproic and butyric acids was performed to produce reduced‐calorie structured lipids (SL). The SL were obtained by incubating a 1:4:4 mole ratio of triolein, caproic acid, and butyric acid, respectively, with 10% of lipase (w/w of total substrates) in 1.5 mL hexane at 55°C for 24 h. Of nine commercially avaialble lipases screened, IM60, which contains the lipase from Rhizomucor miehei, was the most effective and produced 13 mol% unreacted triolein, 49% disubstituted, and 38% monosubstituted triacylglycerols that contained short‐chain fatty acids. The products were analyzed by reverse‐phase high performance liquid chromatography with an evaporative light‐scattering detector. Reaction parameters studied included time course, temperature, enzyme load, and substrate mole ratio. The yields obtained demonstrate that a structured lipid with long‐chain and short‐chain fatty acids can be synthesized by using IM60 lipase in organic medium.

Preparation of S-2-Ethylhexyl-para-methoxycinnamate by lipase catalyzed sequential kinetic resolution

Abstract S-2- Ethylhexyl -para- methoxycinnamate S- 6 is prepared by a sequential biocatalytic resolution. First, either enantioselective acetylation of rac 2-ethylhexanol (±)- 1 by vinylacetate to R -(−)-2-ethylhexylacetate R- 2 , or alcoholysis of rac 2-ethylhexylbutyrate (±)- 3 by n -butanol to S-(+)-2- ethylhexanol S- 1 is completed, than, without isolation of the enantiomerically enreached S -alcohol, its enantioselective acylation with the activated para -methoxycinnammic acid derivatives 4,5 is performed, both steps being catalyzed by different microbial lipases. The highest amplification of enantioselectivity is obtained by combining acetylation of rac 2-ethylhexanol catalyzed by Penicillium camembertii lipase or alcoholysis of rac 2-ethylhexylbutyrate catalyzed by Pseudomonas species lipase in the first step, with acylation of enantiomerically enreached S- 1 by vinyl-para-methoxycinnamate 4 catalyzed by Lipozyme IM lipase;84.5% e.e. of S- 6 is achieved in the first, and 88% e.e. in the second approach. Since the R -enantiomer of 2-ethylhexanol represents potential source of teratogenic R -2-ethylhexanoic acid, S- 6 is regarded as biologicaly safer UV filter as compared to racemic 2-ethylhexyl- para -methoxycinnamate.

ENZYMATIC SYNTHESIS OF STRUCTURED LIPIDS: TRANSESTERIFICATION OF TRIOLEIN AND CAPRYLIC ACID

ABSTRACTStructured lipids were successfully synthesized by lipase‐catalyzed trans‐esterification (acidolysis) of caprylic acid and triolein in nonaqueous medium. Twelve commercially available lipases (10%, w/w substrates) were screened for their ability to form structured lipid by incubating 100 mg triolein and 65.3 mg caprylic acid in 3 ml hexane at 55C for 24 h. The products were analyzed by reverse‐phase high performance liquid chromatography (HPLC) with evaporative light scattering detection. Monocapryloolein was the major component of the products (57.4 mol %) and IM60 lipase from Rhizomucor miehei was the best biocatalyst. Dicapryloolein and triolein contents were 35.4% and 5.3%, respectively. Temperature, mole ratio, time course, incubation media, added water, enzyme load, and substrate concentration were also investigated in this study. The results suggest that it is possible to synthesize structured lipids with lipase as biocatalyst.