Changes in the polyphenoloxidase (PPO) activity and the phenolic content of peaches (Prunus persica cv. Premier) during postharvest ripening were studied. The fruits were stored at 12 or 25C for up to 15 days. The quantity of extractable proteins was maximum at 6-10 days storage at 25 and 12C, coinciding with the onset of the yellowness in the fruits. The PPO activity increased up to the ripening stage, showing a maximum value at 8 days of storage. This was coincident with the maximum degree of browning as evaluated by the absorbance at 440 nm. The amount of total phenolics and chlorogenic acid in the fruits decreased during storage; however, the differences were not significant. The browning potential closely correlated with the enzyme activity, but not with the phenolic content.

Glycosidase activities were detected as detergent‐insoluble after sequential extractions of goat sperm with Triton X‐100. Seventy percent of total β‐glucuronidase activity was found in the detergent‐insoluble fraction. This portion of β‐glucuronidase was resistant to extractions in the presence of 1 M KCl, chaotropic agents, colchicine, or cytochalasin B, being only partially solubilized by 3 M KCl or DNAse I treatment. The treatment with 0.1% sodium deoxycholate was effective, releasing 73% of the enzyme activity. Treating the deoxycholate extract with DNAse I resulted in a change in the elution profile of β‐glucuronidase as judged by gel filtration chromatography. A polyclonal antibody was developed against pancreatic β‐glucuronidase, and the sperm enzyme was strongly inhibited by the IgG fraction of this antibody. Western blot analysis showed that the same protein correspond to both Triton‐soluble and insoluble enzyme. Results demonstrate that β‐glucuronidase is tightly bound to the Triton X‐100 resistant fraction, suggesting that the enzyme is associated to sperm cytoskeleton. J. Exp. Zool. 289:146–152, 2001. © 2001 Wiley‐Liss, Inc.

Affinity chromatography on Concanavalin-A Sepharose, followed by gel filtration and hydrophobic interaction chromatography, permits the isolation of low molecular weight N-glycosidically linked oligomannosidic glycopeptides (MGp) from the autoproteolysis products of human seminal plasma. The monosaccharide composition of MGp showed only mannose, N-acetylglucosamine and a small amount of galactose. Structural studies were carried out by methylation analysis and chromium trioxide oxidation, and results were consistent with the structures accepted for high-mannose N-glycans. MGp was capable of inhibiting the sperm acrosomal exocytosis mediated by sperm-surface receptors. These data suggest that MGp act as a "decapacitation" factor preventing premature sperm exocytosis.

Glycosidase activities were detected as detergent-insoluble after sequential extractions of goat sperm with Triton X-100. Seventy percent of total beta-glucuronidase activity was found in the detergent-insoluble fraction. This portion of beta-glucuronidase was resistant to extractions in the presence of 1 M KCl, chaotropic agents, colchicine, or cytochalasin B, being only partially solubilized by 3 M KCl or DNAse I treatment. The treatment with 0.1% sodium deoxycholate was effective, releasing 73% of the enzyme activity. Treating the deoxycholate extract with DNAse I resulted in a change in the elution profile of beta-glucuronidase as judged by gel filtration chromatography. A polyclonal antibody was developed against pancreatic beta-glucuronidase, and the sperm enzyme was strongly inhibited by the IgG fraction of this antibody. Western blot analysis showed that the same protein correspond to both Triton-soluble and insoluble enzyme. Results demonstrate that beta-glucuronidase is tightly bound to the Triton X-100 resistant fraction, suggesting that the enzyme is associated to sperm cytoskeleton. J. Exp. Zool. 289:146-152, 2001.