Method for late introduction of the (
The presently disclosed subject matter demonstrates that ThnG and ThnQ enzymes encoded by the thienamycin gene cluster in Streptomyces cattleya oxidize the C-2 and C-6 moieties of carbapenems, respectively. ThnQ stereospecifically hydroxylates PS-5 giving N-acetyl thienamycin. ThnG catalyzes sequential desaturation and sulfoxidation of PS-5, giving PS-7 and its sulfoxide. The ThnG and ThnQ enzymes are relatively substrate selective, but give rise to the oxidative diversity of carbapenems produced by S. cattleya , and orthologues likely function similarly in allied streptomyces.
1. A process for preparing a carbapenem of Formula (II):
the process comprising contacting or incubating a carbapenem substrate of Formula (IIa):
with a ThnG enzyme under reaction conditions to produce the carbapenem of Formula (II);
wherein:
R 1 is H;
R 4 is selected from the group consisting of, —NH-glycyl, —NH-beta-alanyl, —NHC(═O)CH 2 CH 2 CH 2 NH 2 , —NH-pantethienic acid amide, an acyl group comprising C 1 -C 10 substituted or unsubstituted linear or branched alkyl, which can further comprise one or more 3-6 member cycloalkyl rings; and
R 3 is H.
2. The process of claim 1 , wherein R 4 is selected from the group consisting of 2-hydroxyacetyl, 2-methoxyacetyl, 3-hydroxypropionyl, 4-hydroxybutanoyl, and 3,4-dihydroxybutanoyl.
3. The process of claim 1 , wherein the carbapenem of Formula (II) is:
4. A process for preparing a carbapenem of Formula (III):
the process comprising contacting or incubating a carbapenem substrate of Formula (IIIa):
with a ThnG enzyme under reaction conditions to produce a carbapenem of Formula (IIIb):
then contacting or incubating the carbapenem of Formula (IIIb) with a ThnG enzyme under reaction conditions to produce a carbapenem of Formula (III);
wherein:
R 1 is H;
R 4 is selected from the group consisting of —NH-glycyl, —NH-beta-alanyl, —NHC(═O)CH 2 CH 2 CH 2 NH 2 , —NH-pantethenic acid amide, an acyl group comprising C 1 -C 10 substituted or unsubstituted linear or branched alkyl, which can further comprise one or more 3-6 member cycloalkyl rings; and
R 3 is H.
5. The process of claim 4 , wherein R 4 is selected from the group consisting of 2-hydroxyacetyl, 2-methoxyacetyl, 3-hydroxypropionyl, 4-hydroxybutanoyl, and 3,4-dihydroxybutanoyl.
6. The process of claim 4 , wherein the carbapenem of Formula (III) is: