STUDIES ON INDUCTION OF HEPARIN-DEGRADING ENZYMES IN FLAVOBACTERIUM HEPARINUM

Five enzymes from Flavobacterium heparinum acting in concert are able to degrade heparin to monosaccharides. These enzymes have been identified as: a glucosaminidase able to degrade heparin to sulfated disaccharides and oligosaccharides; a glycuronidase which hydrolyzes the disaccharides to glucosamine 2,6-disulfate; a sulfamidase and a sulfoesterase able to remove sulfates from the N and O positions of glucosamine 2,6-disulfate and another sulfoesterase able to partially desulfate one of the disaccharides. The glucosaminidase, glycuronidase, and sulfamidase were induced when the bacteria were grown in the presence of heparin. Low activities of these enzymes could also be detected when the bacteria were grown in the absence of heparin. All three enzymes were also induced in the bacteria with the sulfated di-, tetra-, and hexasaccharides derived from heparin. Glucosamine N-sulfate and glucosamine 2,6-disulfate on the other hand were very poor inducers of these enzymes. Time curve studies on the induction of this enzymic system suggested that the sulfated disaccharides were the actual inducers. The induction by the tetrasaccharides, hexasaccharides, and heparin showed a lag period of about 1 hr, suggesting that these compounds were first hydrolyzed to disaccharides by enzymes present in low levels of activity in noninduced Flavobacterium. Actinomycin D, mitomycin C, and chloramphenicol were potent inhibitors of this induction process. © 1969, American Chemical Society. All rights reserved.