Effect of modification of carbohydrate component on properties of glucoamylase.
In this study, we investigated enzymatic deglycosylation of glucoamylase from Aspergillus awamori X 100/D27, a glycoprotein which has two N-linked and about forty short mannose-bearing O-linked sugars per molecule. O-Linked sugars were modified by treatment with alpha-mannosidase and N-linked sugars were removed using endo-beta-N-acetylglucosaminidase F. Analysis of conformational changes following deglycosylation suggests that O-linked sugars essentially contribute to the stabilization of glucoamylase domains. Modification of the carbohydrate component by adding 1-deoxymannojirimycin to the culture medium induced inhibition of alpha-mannosidases involved in the processing, leading to a more complete glycosylation and, consequently, to a higher stability of the enzyme.
Neustroev
K N
KN
Petersburg Nuclear Physics Institute, Gatchina, St. Petersburg, Russian Federation.
Golubev
A M
AM
Firsov
L M
LM
Ibatullin
F M
FM
Protasevich
I I
II
Makarov
A A
AA
D017485:1-Deoxynojirimycin / Q000737:chemistry; D001230:Aspergillus / Q000201:enzymology; D050260:Carbohydrate Metabolism; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000737:chemistry; D004795:Enzyme Stability; D005087:Glucan 1,4-alpha-Glucosidase / Q000737:chemistry / Q000378:metabolism*; D006031:Glycosylation; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008361:Mannosidases / Q000737:chemistry; D008969:Molecular Sequence Data; D011487:Protein Conformation; D013816:Thermodynamics; D043323:alpha-Mannosidase
D002241:Carbohydrates; D017485:1-Deoxynojirimycin; D008361:Mannosidases; D043323:alpha-Mannosidase; D005087:Glucan 1,4-alpha-Glucosidase