Biochemical origins of lactaldehyde and hydroxyacetone in Methanocaldococcus jannaschii.
The biochemical routes for the metabolism of methylglyoxal and the formation of lactaldehyde and hydroxyacetone in Methanocaldococcus jannaschii have been established. The addition of methylglyoxal and NADH, NADPH, F 420H 2, or DTT to a M. jannaschii cell extract stimulated the production of both lactaldehyde and hydroxyacetone. Using appropriately labeled NADH, NADPH, and F 420H 2, hydride transfer was only observed from F 420H 2 to lactaldehyde. It was shown that cell extracts of this Archaea readily catalyzed the F 420H 2-dependent reduction of methylglyoxal to lactaldehyde, a precursor of the lactate found in coenzyme F 420. This conversion was established by measuring the incorporation of deuterium from (5 RS)[5- (2)H 1]F 420H 2 into the C-2 position of the formed lactaldehyde. In vivo generated (5 R)[5- (2)H 1]F 420H 2 was also found to incorporate deuterium into lactaldehyde. The experimental data indicated that the pro- R hydrogen of F 420H 2 was transferred during the reduction. The stereochemistry of this transfer was opposite from that observed for all other known enzyme-catalyzed hydride-transfer reactions involving F 420. [1,3,3,3- (2)H 4]-Methylglyoxal was incorporated into lactaldehyde and hydroxyacetone as an intact unit during this reduction with the occurrence of some deuterium exchange. The exchange observed during this incorporation into lactaldehyde was substantially more than the exchange observed during the incorporation into the hydroxyacetone. The hydroxyacetone was derived directly from methylglyoxal, with the hydrogen for the reduction being derived from water. Hydroxyacetone was also readily formed by the condensation of pyruvate with formaldehyde. The product of the MJ0663 gene was shown to catalyze this condensation reaction.
White
Robert H
RH
Department of Biochemistry, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061, USA. rhwhite@vt.edu
D000095:Acetolactate Synthase / Q000378:metabolism; D000096:Acetone / Q000031:analogs & derivatives* / Q000378:metabolism; D000447:Aldehydes / Q000378:metabolism*; D002457:Cell Extracts; D005557:Formaldehyde / Q000378:metabolism; D007185:Incubators; D017015:Methanococcales / Q000166:cytology / Q000378:metabolism*; D010084:Oxidation-Reduction; D011765:Pyruvaldehyde / Q000378:metabolism; D019289:Pyruvic Acid / Q000378:metabolism; D019163:Reducing Agents / Q000494:pharmacology; D012256:Riboflavin / Q000031:analogs & derivatives / Q000378:metabolism
D000447:Aldehydes; D002457:Cell Extracts; D019163:Reducing Agents; D000096:Acetone; D005557:Formaldehyde; C014634:lactaldehyde; C015084:coenzyme F420; D011765:Pyruvaldehyde; C004433:acetol; D019289:Pyruvic Acid; D000095:Acetolactate Synthase; D012256:Riboflavin