Identification of Oligosaccharides Formed during Stachyose Hydrolysis by Pectinex Ultra SP-L

Abstract

The commercial enzyme preparation Pectinex Ultra SP-L containing fructosyltransferase activity was used to hydrolyze stachyose. During this reaction, besides the formation of mono-, di-, and trisaccharides (DP(3)), the presence of one pentasaccharide (DP(5)) and a new oligosaccharide (DP(6)) has been detected by gas chromatography. DP(5) and DP(6) oligosaccharides were isolated and fully characterized for the first time by an extensive nuclear magnetic resonance (NMR) study. Complete structure elucidation and full proton and carbon assignments were carried out using 1D ((1)H, (13)C) and 2D (gCOSY, multiplicity-edited gHSQC, gHSQC-TOCSY, and gHMBC) NMR experiments. The two oligosaccharides were shown to be stachyose-based structures; the pentasaccharide has a fructose unit linked to the C-1 of the fructose end of stachyose, and the hexasaccharide has a fructose unit linked to the C-1 of the fructose end of the pentasaccharide. The fructosyltransferase activity present in Pectinex Ultra SP-L allows new uses of this commercial enzyme preparation in the synthesis of oligosaccharides derived from alpha-galactosides.

Study of Influential Factors on Oligosaccharide Formation by Fructosyltransferase Activity during Stachyose Hydrolysis by Pectinex Ultra SP-L

“…been previously reported (Montilla et al, 2009). Besides, MALTI-TOF-MS analysis of oligosaccharide-enriched fractions allowed the detection of oligosaccharides with DP7…”

“…Purified fructosyltransferase from Aspergillus niger has been used to catalyse the synthesis of new oligosaccharides from raffinose (Uhm, Baek, Jhon, Kim, Hwang, & Ryu, 1999). The relatively inexpensive commercial enzyme preparation Pectinex Ultra SP-L, produced by Aspergillus aculeatus, was shown to contain fructosyltransferase activity (Hang & Woodams, 1995;Tanriseven & Aslan, 2005;Ghazi et al, 2006;Montilla, Corzo, Olano, & Jimeno, 2009); therefore, it could be used as catalyst in large-scale production of new oligosaccharides. Therefore, the objective of this work has been to investigate in more detail the fructosyltransferase activity from Aspergillus aculeatus of commercial enzymatic preparation Pectinex Ultra SP-L over stachyose hydrolysis.…”

Study on the digestion of milk with prebiotic carbohydrates in a simulated gastrointestinal model

“…Trimethyl silylated oximes (TMSO) of carbohydrates (mono-, di-and trisaccharides) present in samples were determined by Gas Chromatography following the method described by Montilla et al (2009). Samples corresponding to 0.5 mg of saccharides were added to 0.2 mL of Internal Standard (I.S.)…”

Effects of conventional and ultrasound blanching on enzyme inactivation and carbohydrate content of carrots

“…The analysis was performed by GC as described by Soria et al [21] with a gas chromatograph (Agilent Technologies 7890A) equipped with a flame ionization detector (FID) and using nitrogen as carrier gas at a flow rate of 1 mL min -1 . The trimethylsilyl oxime (TMSO) derivatives, prepared as described by Montilla et al [22], were separated using an HP-5MS capillary column (5 % phenyl methylsilicone, 30 m 9 0.25 mm i.d. 9 0.25 lm film thickness; J&W Scientific, Folsom, CA, USA).…”