An In Silico Approach to Enzymatic Synthesis of Fucooligosaccharides Using α-L-Fucosidase from Thermotoga maritima

Abstract: Fucooligosaccharides comprise the primary group of human milk oligosaccharides. Due to their beneficial properties, a series of synthetic methods have been proposed to obtain them. Enzy- matic methods show great promise, and α-L-fucosidase from Thermotoga maritima has emerged as a powerful catalyst for their production. Nonetheless, the enzyme’s limited substrate scope has de- layed its wider application. The present work aims to compare the relative reactivity of fucose, pNP-fucose, and ethyl-fucose, while also exploring the molecular interactions of these fucosyl-donors with the enzyme through a combination DFT and docking analysis. The HOMO-LUMO band gaps range from −7.14571 to −4.24429 eV, with α/β-pNP-fucose and α-fucose being the three most reactive compounds. Moderate association energies between −6.4 to −5.5 kcal·mol−1 were found in the dock- ing analysis, with α-pNP-fucose and both anomers of ethyl-fucose demonstrating the poorest affinity. In the case of α/β-lactose affinity to the β-fucose/enzyme complex, no significant differ- ences were shown. We conclude that the best fucosyl-donors for transfucosylation are those that maintain an enzyme affinity and reactivity similar to pNP-fucose

Efficient Synthesis of 2-Substituted 7-Azaindole Derivatives via Palladium-Catalyzed Coupling and C-N Cyclization Using 18-Crown-6

A practical and straightforward preparation of various novel 2-substituted 7-azaindole derivatives from 2-amino-3-iodopyridine by a two-step procedure is described that gives the desired compounds in good overall yields

Waste residues from Opuntia ficus indica for peroxidase-mediated preparation of phenolic dimeric compounds

A methodology to detect peroxidase activity in Opuntia ficus indica cladodes waste extracts was performed and then used towards phenolic compounds. The extracts were able to dimerize three different molecules. Dimeric compounds were produced with yields ranging from 11% to 55%. The influence of H2O2 concentration was also tested, finding better yields when the peroxide-to-substrate ratio was 1:1. Some water-miscible solvents were used trying to increase overall yields, but no-significant positive results were found. In fact, one of them, THF, seemed to inhibit dimerization reaction. Hence, we have tested an alternative natural peroxidase source obtained from the wastes of a local highly-consumed vegetable and studied their enzymatic activity towards the preparation of biologically active, valuable compounds. Keywords: Opuntia ficus cladodes, Food waste residues, Peroxidase, Oxidative coupling, Phenolic compounds, Dimerization reaction

An In Silico Approach to Enzymatic Synthesis of Fucooligosaccharides Using α-l-Fucosidase from Thermotoga maritima

Fucooligosaccharides comprise the primary group of human milk oligosaccharides. Due to their beneficial properties, a series of synthetic methods have been proposed to obtain them. Enzymatic methods show great promise, and α-l-fucosidase from Thermotoga maritima has emerged as a powerful catalyst for their production. Nonetheless, the enzyme’s limited substrate scope has delayed its wider application. The present work aims to compare the relative reactivity of fucose, pNP-fucose, and ethyl-fucose, while also exploring the molecular interactions of these fucosyl-donors with the enzyme through a combination DFT and docking analysis. The HOMO-LUMO band gaps range from −7.14571 to −4.24429 eV, with α/β-pNP-fucose and α-fucose being the three most reactive compounds. Moderate association energies between −6.4 to −5.5 kcal·mol−1 were found in the docking analysis, with α-pNP-fucose and both anomers of ethyl-fucose demonstrating the poorest affinity. In the case of α/β-lactose affinity to the β-fucose/enzyme complex, no significant differences were shown. We conclude that the best fucosyl-donors for transfucosylation are those that maintain an enzyme affinity and reactivity similar to pNP-fucose

Influence of Oat β-Glucan on the Survival and Proteolytic Activity of Lactobacillus rhamnosus GG in Milk Fermentation: Optimization by Response Surface

β-glucans come from cereals that have been located within compounds with prebiotic activity. They have presented several bioactivities that have determined their high functional value. The aim of this study was to identify the influence of oat β-glucan on the survival and proteolytic activity of Lactobacillus rhamnosus GG in a milk fermentation through an experimental design to optimize the process. For β-glucan extraction after dry milling of oats, two methods were applied: with and without enzymatic inactivation of the semolina. The highest extraction yield (45.25 g/L) was obtained with enzymatic inactivation. For the optimization of survival and proteolytic activity, a central design composed of axial points with two factors on three levels was used. Control factors were β-glucan and inoculum concentrations. According to response surface, the best survival growth rate of probiotic was observed with 4.38% of inoculum and 22.46 g/L of β-glucan, and the highest production of free amino groups was observed with 4.18% of inoculum and 22.71 g/L of β-glucan. Thus, β-glucan promotes the proteolytic activity of Lb. rhamnosus GG in milk fermentation