Preparation of sialylated oligosaccharides employing recombinant trans-sialidase from Trypanosoma cruzi

Abstract

Terminally sialylated oligosaccharides were synthesised employing recombinant trans-sialidase from Trypanosoma cruzi. Regio- and stereoselectively Sia-alpha(2-3)-Gal-betaR derivatives could be obtained in respectable yields, using combined chemical and enzymatic methodologies. An array of different disaccharide precursors such as Gal-beta(1-3)-GalNAc-alphaSer/Thr, lactosides and lactosamide derivatives were sialylated and successfully purified by facile isolation procedures. Depending on the acceptor structure isolated, yields for trans-sialylation products were between 20 and 60%.

Subsequent Enzymatic Galactosylation and Sialylation Towards Sialylated Thomsen–Friedenreich Antigen Components

“…), employing a Gal residue in the acceptor site. [44] In other studies reactions with the immobilized enzyme were investigated. [45] In our further work we could use 2-deoxy-(25) and 6-deoxy derivatives (24) of d-galactose as well as l-arabinose (23) as building block B [46] to investigate the influence of the hydroxy groups in positions 2 and 6 or a missing CH 2 OH group on the transfer employing trans-sialidase.…”

“…Trans-glycosylation with T. cruzi took place with equimolar amounts of substrate and in eight hours incubation time as developed previously. [44] Higher enzyme activities were achieved in a buffer system of Tris-HCl buffer at pH 7.5 and a temperature of 23 8C.…”

Chemical synthesis using enzymatically generated building units for construction of the human milk pentasaccharides sialyllacto-N-tetraose and sialyllacto-N-neotetraose epimer

“…The subsequent transsialylation was carried out with p -nitrophenyl sialoside (pNp-αNeu5Ac) and either sialidase from Salmonella typhimurium or from Newcastle disease virus [7]. Recently, a more effective higher yielding transfer has been reported in which sialylation with recombinant transsialidase ( Trypanosoma cruzi ) gave the trisaccharide 3 in 32% yield [8]. Treatment of 3 with methanol and acidic ion exchange resin led to the methyl ester (for the method cf.…”

“…Further sialylation at position 3′-OH with pNp-αNeu5Ac and either sialidase from Vibrio cholerae or Clostridium perfringens afforded the α,2-6-sialylated trisaccharide 9 exclusively [7]. Later studies showed that 9 could be obtained in an enhanced yield of 32% by transsialylation with recombinant transsialidase ( Trypanosama cruzi ) [8]. Formation of the methyl ester and peracetylation led to the trisaccharide donor building block 10 .…”

Chemoenzymatic Synthesis of Functional Sialyl LewisX Mimetics with a Heteroaromatic Core

“…The title compound was prepared according to a method reported by Neubacher et al16b Azide 8 [26] (400 mg, 1.95 mmol) and 4-nitrophenyl-α-sialoside 9 [27] (492 mg, 1.10 mmol) were dissolved in Tris-HCl-buffer (8 mL, 100 m m , pH 7.9). The mixture was cooled to 13 °C by using a cryostat and an aqueous solution of recombinant trans -sialidase (4 mL, c =0.93 mg mL −1 ) was added.…”