Purification and Glycosylation Analysis of an Acidic Pectin Methylesterase in Jelly Fig (Ficus awkeotsang) Achenes

An acidic pectin methylesterase (PME) is responsible for the gelation of water extract from jelly fig (Ficus awkeotasang) achenes. A new, fast and efficient, method has been developed to purify this acidic PME. The method includes preparing jelly curd by traditional hand washing, extracting proteins...

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Bibliographic Details
Published in:Journal of agricultural and food chemistry 2002-05, Vol.50 (10), p.2920-2925
Main Authors: Ding, Joe L. C, Hsu, Joyce S. F, Wang, Miki M. C, Tzen, Jason T. C
Format: Article
Language:English
Subjects:
Acetylglucosamine - analysis
Agronomy. Soil science and plant productions
Biological and medical sciences
Carboxylic Ester Hydrolases - isolation & purification
Carboxylic Ester Hydrolases - metabolism
Chromatography, Gel
Economic plant physiology
Electrophoresis, Polyacrylamide Gel
Enzymes
Ficus - enzymology
Food additives
Food industries
Fundamental and applied biological sciences. Psychology
Galactose - analysis
General aspects
Glycosylation
Hydrogen-Ion Concentration
Mannose - analysis
Monosaccharides - analysis
Nutrition. Photosynthesis. Respiration. Metabolism
Polysaccharides - analysis
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Summary:An acidic pectin methylesterase (PME) is responsible for the gelation of water extract from jelly fig (Ficus awkeotasang) achenes. A new, fast and efficient, method has been developed to purify this acidic PME. The method includes preparing jelly curd by traditional hand washing, extracting proteins from the curd, and separating PME by anion-exchanger. The purified PME exists as a monomer of 38 kDa determined by gel filtration, and exerts enzymatic activity over a broad pH range, particularly in acidic environments where most known PME enzymes from various species are inactivated. Chemical staining and enzymatic cleavage suggest that the jelly fig PME is an N-linked glycoprotein. Fluorophore-assisted carbohydrate electrophoresis reveals that the polysaccharide of this glycoprotein putatively consists of 22 hexoses including 16 mannose, 4 N-acetylglucosamine, and 2 galactose residues. Keywords: Gelation; jelly fig; N-glycosylation; pectin methylesterase; polysaccharide
ISSN:0021-8561
1520-5118
DOI:10.1021/jf010845+