Study of galacto‐oligosaccharide formation from lactose using Pectinex Ultra SP‐L

Abstract

BACKGROUND: Galacto-oligosaccharides (GOS) are synthesised from lactose by transglycosylation using β-galactosidase (EC 3.2.1.23) and are recognised as prebiotics. The commercial enzyme preparation Pectinex Ultra SP-L produced by Aspergillus aculeatus possesses β-galactosidase activity; however, because its use has been directed towards the formation of 6 -β-galactosyl-lactose, no data have been reported on the formation of other GOS. Since the composition of the oligosaccharide mixture obtained during lactose hydrolysis may affect the prebiotic properties, in this study the influence of various parameters (pH, temperature, time and enzyme and lactose concentrations) on the formation of GOS using Pectinex Ultra SP-L was investigated.

Improvement of covalent immobilization procedure of β‐galactosidase from Kluyveromyces lactis for galactooligosaccharides production: Modeling and kinetic study

“…The kinetics observed in this study is different from the reports of Mart ınez-Villaluenga et al 14 and Kim et al 38 because they were able to synthesize up to trisaccharides, whereas in this investigation, the immobilized enzyme could synthesize up to tetrasaccharides using lactose concentration lower than that reported by Chockchaisawasdee et al 13 It is clear that either the free or immobilized enzyme and its source can affect yield and type of GOS produced, as reported by Cardelle-Cobas et al 42 using b-galactosidase from Aspergillus aculeatus.…”

Analysis, structural characterization, and bioactivity of oligosaccharides derived from lactose

“…HPAEC-PAD profiles of oligosaccharides formed from transgalactosylation of lactulose using a ␤-galactosidase from Aspergillus aculeatus at pH 6.5, 60°C, 650 g/L of lactulose, and 16 units/mL of enzyme after 7 h of reaction. Identified compounds: (1) galactose; (2) Under these conditions, the weakly acidic nature of carbohydrates gives highly selective separations so that a number of studies on the use of HPAEC in the analysis of lactose-derived oligosaccharides have been performed during the last few years [26,[36][37][38][39][40][41][42]. This is a powerful HPLC operating method capable of separating oligosaccharides based on structural features such as size, charge, composition, anomericity, and linkage isomerism [43].…”

“…The analysis of oligosaccharides formed during chemical or enzymatic modifications of lactose is mainly accomplished by high‐performance anion exchange chromatography (HPAEC) using CarboPac® PA columns with sodium hydroxide and sodium acetate solutions as mobile phases. Under these conditions, the weakly acidic nature of carbohydrates gives highly selective separations so that a number of studies on the use of HPAEC in the analysis of lactose‐derived oligosaccharides have been performed during the last few years . This is a powerful HPLC operating method capable of separating oligosaccharides based on structural features such as size, charge, composition, anomericity, and linkage isomerism .…”

“…With respect to detectors for HPLC analysis of lactose‐derived oligosaccharides, UV detector can be applicable only to carbohydrates with UV‐absorbing chromophores such as sialic acid, aldonic acid, or amino residues found in HMO , whereas neutral carbohydrates present in milk require prior derivatization . Among the detection systems that can be coupled to HPAEC, pulsed amperometric detection (PAD) enables the universal and highly sensitive detection, reportedly in the picomole range , of all carbohydrates , being used to analyze mixtures of GOS formed during enzymatic lactose hydrolysis by β‐galactosidases from Lactobacillus reuteri , Lactobacillus plantarum , Kluyveromyces lactis , B. circulans , and Aspergillus aculeatus . HPAEC‐PAD has also been used to quantify levels of GOS in commercial lactose‐free ultra high temperature (UHT) dairy products and commercial fermented milks , as well as to the study of the new oligosaccharides formed during enzymatic hydrolysis of lactulose by β‐galactosidase from K. lactis and A. aculeatus .…”

Síntese de galacto-oligossacarídeos a partir de lactose usando β-galactosidase comercial de Kluyveromyces lactis

“…A formação de GOS a partir da lactose é influenciada por diversos fatores, como a fonte da enzima, a concentração da enzima, a temperatura, o pH e a concentração do substrato (BOON et al, 2000;CARDELLE-COBAS et al, 2008). O planejamento fatorial tem se constituído em poderosa ferramenta para o planejamento e a análise de experimentos em sistemas dependentes de mais de uma variável.…”