Permeabilized probiotic Lactobacillus plantarum as a source of β-galactosidase for the synthesis of prebiotic galactooligosaccharides

Abstract

Permeabilized probiotic Lactobacillus plantarum was used as a source of β-galactosidase for the synthesis of galactooligosaccharides (GOS) from lactose. β-galactosidase activity was highest when galactose (1,724 Miller Units) was used as a carbon source compared to lactose, sucrose or glucose at 37 °C, 18 h. Permeabilized cells had the highest transgalactosylation activity resulting in 34 % (w/w) GOS synthesis from 40 % (w/v) lactose at 50 °C over 12 h. HPLC revealed that the GOS were composed of 13 % disaccharides (non-lactose), 17 % trisaccharides and 4 % tetrasaccharides that were further confirmed by ESI–MS.

Synthesis of Galactosyl Mannitol Derivative Using β-Galactosidase from Kluyveromyces lactis

“…Martínez-Villaluenga et al [2008] using an enzyme from Kluyveromyces lactis obtained the maximum effi ciency of disaccharides at the level of 15.2% in a 35% lactose solution (40 o C, pH 7.5, 5 h), which reached 30% together with trisaccharides. Gobinath & Prapulla [2014] came to a similar fraction profi le (13% of dimers, 17% of trimers) using β-galactosidase from Lactobacillus plantarum in a 40 g/100 mL solution of lactose (pH 7.0, 50 o C, 12 h). A similar level of dimer content was observed after using an 2850 LAU/100g lactose 5700 LAU/100g lactose 11400 LAU/100g lactose 17100 LAU/100g lactose 22800 LAU/100g lactose 28500 LAU/100g lactose FIGURE 1.…”

Transgalactosylating β-galactosidase from probiotic Lactobacillus plantarum MCC2156: production and permeabilization for use as whole cell biocatalyst

“…Lactobacillus plantarum MCC2156, a kanjika isolate was cultured and maintained in Lactobacillus MRS Medium as reported earlier (Gobinath and Prapulla 2014).…”

“…Galactose and yeast extract were chosen as the carbon and nitrogen source, respectively, based on our earlier results (Gobinath and Prapulla 2014). Further, the effect of varying the concentration of the selected medium (M3) components (galactose, yeast extract, sodium acetate, metal ions); initial media pH (4.0-8.0) and incubation temperatures (25-45°C) on β-galactosidase production was also studied.…”

“…The transgalactosylation efficacy of permeabilized biocatalysts for the synthesis of GOS using lactose as substrate was carried out as described earlier (Gobinath and Prapulla 2014). The GOS synthesis was carried out by incubating lactose (40 %, w/v) in 0.05 M sodium phosphate buffer (pH 7.0) with 10 U of biocatalyst/ ml of reaction mixture at 50°C, in a shaking water bath (100 rpm) for 12 h. The reaction was arrested by heating the reaction mixture for 10 min, at 95°C.…”

“…Among various sources of β-galactosidases, intracellular β-galactosidase from probiotic lactic acid bacteria (LAB) such bifidobacteria and lactobacilli for lactose hydrolysis and GOS synthesis is of great industrial relevance mainly due to the 'generally recognized as safe (GRAS)' status of LAB (Hsu et al 2007;Splechtna et al 2006). Intracellular β-galactosidase from probiotic Lactobacillus plantarum MCC2156 exhibited potent transgalactosylating efficacy for the synthesis of prebiotic GOS (Gobinath and Prapulla 2014). Downstream processing of intracellular enzymes involves cell disruptions, which could lead to low yields and decreased activity due to harsh conditions, additionally, adds to the cost of purification processes.…”