Abstract
The enzyme β-galactosidase (β-gal) has extensively used for improvement of lactose intolerance condition. Present study, was designed to assess the potential of β-gal enzyme produced by Lactobacillus fermentum, a kefir isolate, as a biocatalyst for the manufacture of prebiotic galactooligosaccharides (GOS) from lactose. The efficiency of L. fermentum to produce β-gal of 4,254 u/ml was determined by permeabilizing the cells with solvents such as sodium dodecyl sulfate (SDS) and chloroform. Different parameters contributing β-gal production including reaction time, temperature, pH, carbohydrates, and substrate concentration on L. fermentum were studied and optimum β-gal activity was found to be 6,232.13 u/ml. It was observed that different experimental parameters for pH (7.0), temperature (35°C), and carbohydrates (galactose) were statistically significant (p<0.05). L. fermentum was found to produce GOS by transgalactosylation catalysed by β-gal during lactose hydrolysis which yielded di, tri, and tetra oligosaccharides, confirmed by TLC and HPLC. The culture showed β-gal activity, suggesting biotechnological applications and a promising organism for industrial β-gal production.
Highlights
The enzyme β-galactosidase (β-gal) has extensively used for improvement of lactose intolerance condition
Microorganisms are considered as suitable source for β-gal production in industries because of its high multiplication rate, yield and stability. β-gal producing lactic acid bacteria (LAB) including L. plantarum MCC2156, L. reuteri L103 and L461, L. acidophilus R22, L. thermophilus, L. crispatus ATCC 33820, and L. delbrueckii spp. bulgaricus ATCC 11842 were reported (Gobinath and Prapulla, 2014; Kim and Rajagopal, 2000; Nguyen et al, 2006; Nguyen et al, 2007) and are predominantly present in brush border of small intestine
In turn to assess the efficacy of the L. fermentum MT468249, bacterial growth in mid-logarithmic phase was used and plated with different dilutions (10-5, 10-6 and 107)
Summary
The enzyme β-galactosidase (β-gal) has extensively used for improvement of lactose intolerance condition. Present study, was designed to assess the potential of β-gal enzyme produced by Lactobacillus fermentum, a kefir isolate, as a biocatalyst for the manufacture of prebiotic galactooligosaccharides (GOS) from lactose. Different parameters contributing β-gal production including reaction time, temperature, pH, carbohydrates, and substrate concentration on L. fermentum were studied and optimum β-gal activity was found to be 6,232.13 u/ml. L. fermentum was found to produce GOS by transgalactosylation catalysed by β-gal during lactose hydrolysis which yielded di, tri, and tetra oligosaccharides, confirmed by TLC and HPLC. GOS has a low calorific value of about 1.7 k cal/g and can be synthesized using lactose as substrate in the presence of β-galactosidase (β-gal) enzyme. Β-gal known as lactases, are of industrial importance They are produced by a number of sources including microorganisms, plants, animals, vegetables and recombinant sources Β (1→4) glycosidic linkage of lactose (galactosyl β-(1→4) glucose) is splited into its monosaccharides i.e., glucose and galactose, whereas in transgalactosylation, galactose is transferred to lactose rather than transferring to its hydroxyl group to form di, tri or tetra saccharides and form higher degree of polymerization (Panesar et al, 2007)
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