Degradation of monogalactosyl diglyceride and digalactosyl diglyceride by sheep pancreatic enzymes.
1. Saline extract of sheep pancreas acetone-dried powder was shown to catalyse acyl ester hydrolysis of spinach leaf galactosyl diglycerides and also galactosylglucosyl diglyceride of Lactobacillus casei. 2. Sodium deoxycholate stimulated the enzyme activity. Ca(2+) had no effect on the hydrolysis of monogalactosyl diglyceride, but it enhanced that of digalactosyl diglyceride. When added together, there was considerably less activity with both the substrates. 3. Optimal hydrolysis was observed at pH7.2. 4. The initial point of hydrolysis was at position-1, leading to the formation of monogalactosyl monoglyceride and digalactosyl monoglyceride. Further hydrolysis to the corresponding galactosylglycerols and later to galactose and glycerol was also observed, indicating the presence of alpha- and beta-galactosidases in the enzyme preparation. 5. Formation of monogalactosyl diglyceride from digalactosyl diglyceride by the action of alpha-galactosidase was noted. 6. Monogalactosyl diglyceride was also hydrolysed by beta-galactosidase to a limited extent, giving rise to diacylglycerol and galactose. 7. Attempts at purification of monogalactosyl diglyceride acyl hydrolase by using protamine sulphate treatment, Sephadex G-100 filtration and DEAE-cellulose chromatography gave a partially purified enzyme which showed 9- and 81-fold higher specific activity towards monogalactosyl diglyceride and digalactosyl diglyceride respectively. This still showed acyl ester hydrolysis activity towards methyl oleate, phosphatidylcholine and triacylglycerol. 8. When sheep, rat and guinea-pig tissues were compared, guinea-pig tissues showed the highest activity towards both monogalactosyl diglyceride and digalactosyl diglyceride. In all the species pancreas showed higher activity than intestine.
D000818:Animals; D002118:Calcium; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004075:Diglycerides / Q000378:metabolism*; D005696:Galactosidases / Q000378:metabolism; D005989:Glycerides / Q000378:metabolism*; D006168:Guinea Pigs; D006863:Hydrogen-Ion Concentration; D006867:Hydrolases / Q000302:isolation & purification; D006868:Hydrolysis; D007422:Intestines / Q000201:enzymology; D009829:Oleic Acids / Q000378:metabolism; D010179:Pancreas / Q000201:enzymology*; D010713:Phosphatidylcholines / Q000378:metabolism; D051381:Rats; D012756:Sheep; D013997:Time Factors
D004075:Diglycerides; D005989:Glycerides; D009829:Oleic Acids; D010713:Phosphatidylcholines; D006867:Hydrolases; D005696:Galactosidases; D002118:Calcium