Cytosolic Pyridoxine-β-d-Glucoside Hydrolase from Porcine Jejunal Mucosa

During studies of the nutritional utilization of pyridoxine 5′-β-d-glucoside, a major form of vitamin B6 in plants, we detected two cytosolic β-glucosidases in jejunal mucosa. As expected, one was broad specificity β-glucosidase that hydrolyzed aryl β-d-glycosides but not pyridoxine β-d-glucoside. We also found a previously unknown enzyme, designated pyridoxine-β-d-glucoside hydrolase, that efficiently hydrolyzed pyridoxine β-d-glucoside. These were separated and purified as follows: broad specificity β-glucosidase 1460-fold and pyridoxine-β-d-glucoside hydrolase 36,500-fold. Purified pyridoxine-β-d-glucoside hydrolase did not hydrolyze any of the aryl glycosides tested but did hydrolyze cellobiose and lactose. Pyridoxine-β-d-glucoside hydrolase exhibited a pH optimum of 5.5 and apparent molecular mass of 130 kDa by SDS-polyacrylamide gel electrophoresis and 160 kDa by nondenaturing gel filtration, in contrast to 60 kDa for native and denatured broad specificity β-glucosidase. Glucono-δ-lactone was a strong inhibitor of both enzymes. Ionic and nonionic detergents were inhibitory for each enzyme. Conduritol B epoxide, a potent inhibitor of lysosomal acid β-glucosidase, inhibited pyridoxine-β-d-glucoside hydrolase but not broad specificity β-glucosidase, but both were inhibited by the mechanism-based inhibitor 2-deoxy-2-fluoro-β-d-glucosyl fluoride. Our findings indicate major differences between these two cytosolic β-glucosidases. Studies addressing the role of vitamin B6 nutrition in regulating the activity and its consequences regarding pyridoxine glucoside bioavailability are in progress.

• Publication year

  1997

• Venue

  Journal of Biological Chemistry

• Publication date

  1997-12-19

• Fields of study

  Biology, Agricultural and Food Sciences, Chemistry

• Identifiers
  DOI 10.1074/jbc.272.51.32025
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar

• No claims are published for this paper.

• No concepts are published for this paper.

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