Optimization of β-galactosidase Production by Batch Cultures of Lactobacillus leichmannii 313 (ATCC 7830™)

Abstract

The endoenzyme β-galactosidase (β-d-galactoside galactohydrolase; EC 3.2.1.23) has been used at industrial scales for the preparation of lactose-free milk and for the conversion of lactose to galacto-oligosaccharides (GOS) prebiotics. In this study, using Plackett-Burman (PB) design and the response surface methodology (RSM), the batch growth conditions for the production of β-galactosidase in DeMan-Rogosa-Sharpe (MRS) media have been studied and optimized for Lactobacillus leichmannii 313 (ATCC 7830™) for the first time. The incubation temperature (30 < T < 55 • C), starting pH (5.5 < pH < 7.5), and carbon source (glucose, lactose, galactose, fructose, and sucrose) were selected as the significant variables for optimization. The maximum crude β-galactosidase production (measured by specific activity) was 4.5 U/mg proteins and was obtained after 12 h of fermentation. The results of the PB design and further optimization by RSM showed that the initial pH of 7.0 and 15.29 g/L of lactose were the levels that gave the optimum observed and predicted β-galactosidase activities of 23.13 U/mg and 23.40 U/mg, respectively. Through RSM optimization, β-galactosidase production increased significantly (over five-fold) in optimized medium (23.13 U/mg), compared with unoptimized medium (4.5 U/mg). Moreover, the crude enzyme obtained was able to hydrolyze lactose and also produce galacto-oligosaccharides. Because its ability to produce β-galactosidase was significantly improved through optimization by RSM, L. leichmannii 313 can serve as a potential source of β-galactosidase for food applications at an industrial scale.

Antioxidant and Anti-Inflammatory Properties of Recombinant Bifidobacterium bifidum BGN4 Expressing Antioxidant Enzymes

“…Hwang found that the growth of BGN4 reached the early stationary phase at that time [ 19 ]. This result is consistent with previous studies indicating that the Fe-dependent SOD activity of a recombinant Pseudomonas putida strain was high in the log-growth and stationary phases of bacterial cells grown in a medium supplemented with FeCl 3 [ 20 ] and the β-galactosidase activity of Lactobacillus leichmannii 313 was the highest in the early stationary phase [ 21 ].…”

Extraction of B-galactosidase Enzyme from Lacticacid Bacteria

“…The best 1262 (2023) 062018 IOP Publishing doi:10.1088/1755-1315/1262/6/062018 4 carbon source to encourage the growth of the of β-galactosidase. This is due to the fact that lactose, specifically the allolactose isomer, stimulates the lacZ gene's translation by binding to the Lac repressor protein and blocking it from activating the operator region of the lac operon [15]. These findings are consistent with earlier research.…”

“…Numerous studies have documented how carbon sources have a significant impact on the synthesis of -galactosidase by different bacteria. Conclusion: Depending on the microorganisms utilized, the importance of carbon supply in this biosynthesis may vary significantly [15]. This study investigated a variety of carbohydrates (glucose and lactose) to determine the optimal carbon source and best sugar for -galactosidase production.…”

Uses of the Response Surface Methodology for the Optimization of Agro-Industrial Processes

“…AI ST Lactose/β-galactosidase CCD pH 7.06 and 15.3 g/L lactose [80] An eco-friendly pressure liquid extraction method to recover anthocyanins from broken black bean hulls.…”