Fusako Kawai

Studies on the microbial formation of sugar derivatives of D-pantothenic acid: I. Isolation and identification of a new derivative of pantothenic acid: 4′(2′)-O-(α--glucopyranosyl)--pantothenic acid

Abstract A new derivative of pantothenic acid was found in a reaction mixture of Sporobolomyces coralliformis IFO 1032 incubated in a medium containing maltose and D -pantothenic acid. This new compound was isolated from an incubation mixture by active charcoal, Amberlite CG-50, Sephadex G-10 and DEAE-cellulose column chromatography, and by paper chromatography. Then, the compound was characterized as 4′(2′)-O-(α- D -glucopyranosyl)- D -pantothenic acid by various analytical methods, including bioautography, paper chromatography and infrared and NMR spectroscopy.

Studies on Transglycosidation to Vitamin B 6 by Microorganisms:Part IV. Purification of a Bacterial Enzyme Catalyzing Pyridoxine Glucoside Synthesis

The enzyme activity to synthesize pyridoxine glucoside was demonstrated in intact cells and cell extracts of genera, Sarcina and Micrococcus. The isolated and identified strain, Micrococcus sp. No. 431 was found to have high activity of this enzyme in its cell extract.The enzyme activity reached to a maximum after 20 hr of cultivation.The enzyme which synthesized pyridoxine glucoside via transglucosidation from sucrose to pyridoxine was purified from Micrococcus sp. No. 431 by means of ammonium sulfate fractionation, DEAE-Sephadex, hydroxylapatite and Sephadex G–100 column chromatographies. The enzyme was purified about 354–fold and confirmed to be homogenous in polyacrylamide-gel electrophoresis and ultracentrifugation.

Microbial synthesis of sugar derivatives of D-pantothenic acid.

Publisher Summary This chapter examines the microbial synthesis of sugar derivatives of D-pantothenic acid. The compound is hydrolyzed by almond β-glucosidase, cellulases type II and type III, and naringinase but not by yeast α-glucosidase. The enzymic hydrolysis of the compound releases pantothenic acid (PaA) and glucose, which are identified by bioautography and paper chromatography. The molar ratio of the PaA and glucose moieties is approximately l:l as determined by the glucostat method, the anthrone method, and bioassay. An α-glucoside of PaA (PaA-α-G) is formed by several microbial cells or α-glucosidases from PaA and maltose. The washed cells of Sporobolomyces coralliformis IFO 1032 forms large amounts of the derivative—PaA-α-G—under optimal conditions. The derivative is separated from PaA by paper chromatography and confirmed by bioautography. The derivative is quantitatively assayed as PaA by microbiological assay after hydrolysis by yeast α-glucosidase.

Enzymatic Formation of d-Pantothemc Acid-β-Glucoside by Various β-Glucosidases

A β-gIucoside of d-pantothenic acid was formed from d-pantothenic acid and β-glucosyl donors such as cellobiose, phenyl-β-d-glucoside, salicin, and 4-methylumbelliferyl-β-d-glucoside and naphthol AS-BI-β-d-glucoside by various β-glucosidases, i.e., almond β-glucosidase, cellulase type II and III, naringinase, and hesperiginase. The compound was isolated from a reaction mixture of almond β-glucosidase by treatment with active charcoal, Amberlite CG–50, and DEAH-cellulose column chromatography, paper chromatography, and Sephadex G-IO gel filtration. Then, the compound was characterized as 4′-O-(β-d-glucopyranosyl)-d-pantothenic acid by various analytical methods including bioassay, paper chromatography, NMR and specific optical rotation. The microbiological activities of the compound were also determined.