Enzymes for the NADPH-dependent reduction of dihydroxyacetone and D-glyceraldehyde and L-glyceraldehyde in the mould Hypocrea jecorina.
The mould Hypocrea jecorina (Trichoderma reesei) has two genes coding for enzymes with high similarity to the NADP-dependent glycerol dehydrogenase. These genes, called gld1 and gld2, were cloned and expressed in a heterologous host. The encoded proteins were purified and their kinetic properties characterized. GLD1 catalyses the conversion of d-glyceraldehyde and l-glyceraldehyde to glycerol, whereas GLD2 catalyses the conversion of dihydroxyacetone to glycerol. Both enzymes are specific for NADPH as a cofactor. The properties of GLD2 are similar to those of the previously described NADP-dependent glycerol-2-dehydrogenases (EC 1.1.1.156) purified from different mould species. It is a reversible enzyme active with dihydroxyacetone or glycerol as substrates. GLD1 resembles EC 1.1.1.72. It is also specific for NADPH as a cofactor but has otherwise completely different properties. GLD1 reduces d-glyceraldehyde and l-glyceraldehyde with similar affinities for the two substrates and similar maximal rates. The activity in the oxidizing reaction with glycerol as substrate was under our detection limit. Although the role of GLD2 is to facilitate glycerol formation under osmotic stress conditions, we hypothesize that GLD1 is active in pathways for sugar acid catabolism such as d-galacturonate catabolism.
Liepins
Janis
J
VTT Biotechnology, Espoo, Finland.
Kuorelahti
Satu
S
Penttilä
Merja
M
Richard
Peter
P
D003001:Cloning, Molecular; D004098:Dihydroxyacetone / Q000378:metabolism*; D005985:Glyceraldehyde / Q000378:metabolism*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics* / Q000378:metabolism; D035901:Hypocrea / Q000201:enzymology* / Q000235:genetics; D009249:NADP / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013401:Sugar Alcohol Dehydrogenases / Q000235:genetics* / Q000378:metabolism
D011994:Recombinant Proteins; D005985:Glyceraldehyde; D009249:NADP; D013401:Sugar Alcohol Dehydrogenases; C020673:glycerol 2-dehydrogenase (NADP+); D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D004098:Dihydroxyacetone