Chemoenzymatic synthesis of 2-chloro-4-nitrophenyl β-maltoheptaoside acceptor-products using glycogen phosphorylase b

Lili Kandra, Gyöngyi Gyémánt, Magda Pál, Marianna Petró, Judit Remenyik, András Lipták

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Abstract

In the present work, we aimed at developing a chemoenzymatic procedure for the synthesis of β-maltooligosaccharide glycosides. The primer in the enzymatic reaction was 2-chloro-4-nitrophenyl β-maltoheptaoside (G7-CNP), synthesised from β-cyclodextrin using a convenient chemical method. CNP-maltooligosaccharides of longer chain length, in the range of DP 8-11, were obtained by a transglycosylation reaction using α-D-glucopyranosyl-phosphate (G-1-P) as a donor. Detailed enzymological studies revealed that the conversion of G7-CNP catalysed by rabbit skeletal muscle glycogen phosphorylase b (EC 2.4.1.1) could be controlled by acarbose and was highly dependent on the conditions of transglycosylation. More than 90% conversion of G7-CNP was achieved through a 10:1 donor-acceptor ratio. Tranglycosylation at 37 °C for 30 min with 10 U enzyme resulted in G8→12-CNP oligomers in the ratio of 22.8, 26.6, 23.2, 16.5, and 6.8%, respectively. The reaction pattern was investigated using an HPLC system. The preparative scale isolation of G8→11-CNP glycosides was achieved on a semipreparative HPLC column. The productivity of the synthesis was improved by yields up to 70-75%. The structures of the oligomers were confirmed by their chromatographic behaviours and MALDI-TOF MS data.

Original languageEnglish
Pages (from-to)129-136
Number of pages8
JournalCarbohydrate Research
Volume333
Issue number2
DOIs
Publication statusPublished - Jul 3 2001

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Keywords

  • Chain elongation
  • Glycogen phosphorylase b
  • Maltooligosaccharide series
  • Transglycosylation

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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