Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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In Vitro Glycosylation of Peptide (RKDVY) and RNase A by PNGase F

  • Park, Su-Jin (School of Chemical Engineeiring and Institute of Chemical Processes, Seoul National University) ;
  • Lee, Ji-Youn (School of Chemical Engineering and Institute of Chemical Processes, Seoul National University) ;
  • Park, Tai-Hyun (School of Chemical Engineering and Institute of Chemical Processes, Seoul National University)
  • Published : 2003.04.01
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Abstract

The in vitro glycosylation of pentapeptide (Arg-Lys-Asp-Val-Tyr; RKDVY) and RNase A was carried out using PNGase F (peptide-N-glycosidase F), and the results were analyzed using MALDI-TOF-MS. Aminated N,N-diretyl chitobiose was used as the sugar in the glycosylation reaction, and the amination yield of N,N'-diacetyl chitobiose was about $60\%$. To reduce the water activity and shift the reaction equilibrium to a reverse reaction, 1,4-dioxane or ethylene glycol was used as the organic solvent in the enzymatic glycosylation. A certain extent of nonenzymatic glycosylaton, known as the Maillard reaction, was also observed, which occurs on an arginine or lysine residue when the length of tie sugar residue is one or two. However, the extent of glycosylation was much higher in the enzymatic reaction, indicating that PNGase F can be effectively used to produce glycopeptides and glycoproteins in vitro.

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