Fourier transform infrared spectroscopy analysis of Lactose hydrolysis by beta-galactosidase from Lactiplantibacillus plantarum GV54 and Lactiplantibacillus sp. GV66

Abstract

Introduction and Aim: The enzyme beta-galactosidase is extremely important in the food and pharmaceutical industries. The goal of this work was to improve the lactose hydrolysis process employing beta-galactosidase to convert glucose and galactose for alleviating lactose intolerance. Methods: Intracellular beta-galactosidase was extracted from Lactiplantibacillus spp. (GV54 and GV66) by cell permeabilized method using toluene/acetone solvents. Lactose hydrolysis was performed using 50 g/L of lactose with partially purified beta-galactosidase enzyme at different time intervals 2, 4, 6 and 12 h. Fourier transforms infrared spectroscopy (FTIR) was used to evaluate the degree of hydrolysis at different time intervals and lactose hydrolysis was high at 6 h of incubation. Results: The FTIR peak of lactose molecules hydrolysis by beta-galactosidase (GV54 and GV66) with different time intervals (2h, 4h, 6h, and 12h) indicates similarities in bond lengths and vibrational modes. The amide I group of beta-galactosidase is uniformly bound to lactose at 1634 per cm and C-H group of glucose and galactose was observed at 1386 per cm. In the range of 2000-800 per cm, we could identify variations in the shape and strength of these peaks, compared with control (lactose solution). Conclusion: The precise and direct measurement of individual carbohydrates such as glucose and galactose in lactose solution hydrolyzed by beta-galactosidase enzyme was successfully accomplished using FTIR spectrometry. Comparing FTIR analysis to traditional procedures, the time required is drastically decreased. These findings have the analytical power to follow the dynamics of in vitro enzymatic activity of naturally occurring substrates rich in natural carbohydrate polymers.