Method for the production of mucin-type glycopeptide
AssigneeIkuko Ishii Karakasa
InventorsIkuko Ishii Karakasa, Katsumi Ajisaka
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Abstract
Methods for the production of a mucin-type glycopeptide comprising a transglycosylation using a sugar acceptor such as peptide and a sugar donor as an oligosaccharide with an endo-α-N-acetylgalactosaminidase under a given condition is disclosed. The method provides a new practical way to produce mucin-type glycopeptides in industry and can provide an sufficient amount of the mucin-type glycopeptides in a practical use.
Claims16
A method for the production of mucin-type glycopeptide comprising a transglycosylation using a sugar donor and a sugar acceptor with an endo-α-N-acetylgalactosaminidase wherein the endo-α-N acetylgalactosaminidase is produced by a microorganism which is a Streptomyces spp.
A method for the production of mucin-type glycopeptide comprising a transglycosylation using a sugar donor and a sugar acceptor with an endo-α-N-acetylgalactosaminidase wherein the endo-α-N-acetylgalactosaminidase is produced by a microorganism which is a Streptomyces spp., and further wherein the endo-α-N-acetylgalactosaminidase has the following physicochemical properties:
- (a) Action:
- acting on p-nitrophenyl-alpha-N acetylgalactosamine having an alpha-glycoside bond, to release N-acetylgalactosamine and acting on a mucin to release an oligosaccharide having at least two monosaccharides and N-acetylgalactosamine at reducing end;
- (b) Optimum pH:
- optimally active at a pH of about 5 to about 6;
- (c) pH Stability:
- most stable at a pH of about 5.5;
- (d) Optimum temperature:
- optimally active at a temperature ranging from 35 to 40 degree Celsius in a thirty minute reaction;
- (e) Molecular weight:
- about 105,000 determined by gel filtration using Sephadex G-200.
A method for the production of mucin-type glycopeptide comprising a transglycosylation using a sugar donor and a sugar acceptor with an endo-α-N-acetylgalactosaminidase wherein the endo-α-N acetylgalactosaminidase is produced by a microorganism which is a Streptomyces spp., and further wherein the endo-α-N-acetylgalactosaminidase has the following physicochemical properties:
- (a) Action:
- acting on p-nitrophenyl-alpha-N acetylgalactosamine having an alpha-glycoside bond, to release N-acetylgalactosamine and acting on a mucin to release an oligosaccharide having at least two monosaccharides and N-acetylgalactosamine at reducing end;
- (b) Optimum pH:
- optimally active at a pH of about 5 to about 6 when reacted at 37 degree Celsius;
- (c) pH Stability:
- most stable at a pH of about 5.5 and having more than 50% of the maximum activity at a pH ranging from 4.5 to 8.0;
- (d) Optimum temperature:
- optimally active at a temperature ranging from 35 to 40 degree Celsius in a thirty minute reaction and retaining more than 50% of the maximum activity at a temperature ranging from 35 to 55 degree Celsius;
- (e) Thermostability:
- stable at less than 40 degree Celsius;
- (f) Molecular weight:
- about 105,000 determined by gel filtration using Sephadex G-200;
- (g) Effects of metal ions and substances:
- no activity inhibition by Ca2+, Mg2+, and Mn2+ and by EDTA but activity inhibition by PCMB.
Dependencies are inferred from the claim text.