Fourier-transformed infrared assay for the determination of positional specificity of non-specific and 1,3-specific lipases

FARÍAS R; CHEHÍN R; CANELA R

BIOTECHNOLOGY LETTERS

Springer Publishers

Año: 2000 vol. 22 p. 905 - 908

0141-5492

Lipases can be classified in terms of specificity toward the position of the acyl group of triacylglycerols into two groups: 1,3-specific and non-specific. The positional specificity of the lipases is usually determined by TLC (Commenil et al. 1995, Li-Zu-Yi & Ward 1993, Mozaffar & Weete 1993), HPLC (Ergan & Andre 1989, Veeraragavan 1990, Uzawa et al. 1993) or GC (Bereuter & Lorbeer 1995) analysis of the reaction products obtained after various reaction times with triacylglycerols as substrate. In the present paper we report a new, simple and fast infrared spectroscopy method for the characterization of both 1,3-specific and non-specific lipases using a thioester analogue of triacylglycerol: 2,3-dimercapto-1-propanol tributyrate. The method is based on the analysis of specific infrared bands that show the ester and thioester groups of the thioester analogue. It represents an alternative to another spectrophotometric method already described for our group with the same purpose (Farias et al. 1997).et al. 1995, Li-Zu-Yi & Ward 1993, Mozaffar & Weete 1993), HPLC (Ergan & Andre 1989, Veeraragavan 1990, Uzawa et al. 1993) or GC (Bereuter & Lorbeer 1995) analysis of the reaction products obtained after various reaction times with triacylglycerols as substrate. In the present paper we report a new, simple and fast infrared spectroscopy method for the characterization of both 1,3-specific and non-specific lipases using a thioester analogue of triacylglycerol: 2,3-dimercapto-1-propanol tributyrate. The method is based on the analysis of specific infrared bands that show the ester and thioester groups of the thioester analogue. It represents an alternative to another spectrophotometric method already described for our group with the same purpose (Farias et al. 1997).