Ether-cleaving enzyme and diol dehydratase involved in anaerobic polyethylene glycol degradation by a new Acetobacterium sp.
A strictly anaerobic, homoacetogenic bacterium was enriched and isolated from anoxic sewage sludge with polyethylene glycol (PEG) 1000 as sole source of carbon and energy, and was assigned to the genus Acetobacterium on the basis of morphological and physiological properties. The new isolate fermented ethylene glycol and PEG's with molecular masses of 106 to 1000 to acetate and small amounts of ethanol. The PEG-degrading activity was not destroyed by proteinase K treatment of whole cells. In cell-free extracts, a diol dehydratase and a PEG-degrading (ether-cleaving) enzyme activity were detected which both formed acetaldehyde as reaction product. The diol dehydratase enzyme was oxygen-sensitive and was stimulated 10-14 fold by added adenosylcobalamine. This enzyme was found mainly in the cytoplasmic fraction (65%) and to some extent (35%) in the membrane fraction. The ether-cleaving enzyme activity reacted with PEG's of molecular masses of 106 to more than 20000. The enzyme was measurable optimally in buffers of high ionic strength (4.0), was extremely oxygen-sensitive, and was inhibited by various corrinoids (adenosylcobalamine, cyanocobalamine, hydroxocobalamine, methylcobalamine). This enzyme was found exclusively in the cytoplasmic fraction. It is concluded that PEG is degraded by this bacterium inside the cytoplasm by a hydroxyl shift reaction, analogous to a diol dehydratase reaction, to form an unstable hemiacetal intermediate. The name polyethylene glycol acetaldehyde lyase is suggested for the responsible enzyme.
Lehrstuhl Mikrobiologie I Eberhard-Karls-Universität, Tübingen, Germany.
E
E
Schramm
B
B
Schink
D000693:Anaerobiosis; D001673:Biodegradation, Environmental; D003593:Cytoplasm / Q000378:metabolism; D004987:Ethers / Q000378:metabolism; D006094:Gram-Positive Bacteria / Q000145:classification / Q000302:isolation & purification / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000378:metabolism*; D011408:Propanediol Dehydratase / Q000378:metabolism; D012722:Sewage
D004987:Ethers; D012722:Sewage; D011092:Polyethylene Glycols; D011408:Propanediol Dehydratase