In vitro synthesis and purification of UDP-( sup 14 C) galacturonate

Journal Article · Sat Apr 01 04:00:00 EST 1989 · Plant Physiology, Supplement; (USA)

OSTI ID:6610884

Mitcham, E J ^[1]; Gross, K C ^[2]; Wasserman, B P ^[3]

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1. Univ. of Maryland, College Park (USA)
2. Crop Quality Lab., Beltsville, MD (USA)
3. Rutgers Univ., New Brunswick, NJ (USA)

Pectins comprise a major component of the cell wall and much research has focused on degradation of pectins during ripening and senescence. However, little research has been conducted on pectin synthesis, partly due to a lack of commercial availability of UDP-({sup 14}C)galacturonic acid for use as a substrate in assaying galacturonan synthase. We report on the modification and integration of several procedures to synthesize UDP-({sup 14}C) galacturonic acid from commercially available UDP-({sup 14}C)glucuronic acid. A microsomal pellet containing UDP-D-glucuronate-4-epimerase was extracted from 5-day-old mung bean hypocotyls (Phaseolus aureus) and radish roots (Raphanus sativus L.) by ultracentrifugation at 30,000 rpm for 50 min. The UDP-({sup 14}C)galacturonic acid produced was separated from remaining UDP-({sup 14}C)glucuronic acid and other products by electrophoresis in pyridine:acetate:H{sub 2}O on silica gel. Spots were detected by autoradiography, eluted with 80% ethanol, and purified using anion exchange chromatography.

• OSTI ID: 6610884
• Journal Information: Plant Physiology, Supplement; (USA), Journal Name: Plant Physiology, Supplement; (USA) Vol. 89:4; ISSN PPYSA; ISSN 0079-2241
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALDEHYDES
AUTORADIOGRAPHY
CARBON 14 COMPOUNDS
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CHEMICAL PREPARATION
CHROMATOGRAPHY
ELECTROPHORESIS
ENZYMES
FOOD
FRACTIONATION
GALACTURONIC ACID
GLUCURONIC ACID
HYDROXY ACIDS
IN VITRO
ION EXCHANGE CHROMATOGRAPHY
LABELLED COMPOUNDS
LEGUMINOSAE
MAGNOLIOPHYTA
MAGNOLIOPSIDA
MICROSOMES
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANOIDS
PHASEOLUS
PLANTS
RADISHES
RIBOSOMES
SEPARATION PROCESSES
SYNTHESIS
TRANSFERASES
VEGETABLES