Biosynthesis of terpenoids: efficient multistep biotransformation procedures affording isotope-labeled 2C-methyl-D-erythritol 4-phosphate using recombinant 2C-methyl-D-erythritol 4-phosphate synthase.
This paper describes the recombinant expression of the ispC gene of Escherichia coli specifying 2C-methyl-D-erythritol 4-phosphate synthase in a modified form that can be purified efficiently by metal-chelating chromatography. The enzyme was used for the preparation of isotope-labeled 2C-methyl-D-erythritol 4-phosphate employing isotope-labeled glucose and pyruvate as starting materials. The simple one-pot methods described afford numerous isotopomers of 2C-methyl-D-erythritol 4-phosphate carrying (3)H, (13)C, or (14)C from commercially available precursors. The overall yield based on the respective isotope-labeled starting material is approximately 50%.
Hecht
S
S
Lehrstuhl für Organische Chemie und Biochemie, Technische Universität München, Lichtenbergstrasse 4, D-85747 Garching, Germany.
Wungsintaweekul
J
J
Rohdich
F
F
Kis
K
K
Radykewicz
T
T
Schuhr
C A
CA
Eisenreich
W
W
Richter
G
G
Bacher
A
A
D019747:Aldose-Ketose Isomerases / Q000737:chemistry / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D001711:Biotransformation; D002250:Carbon Radioisotopes / Q000378:metabolism; D004896:Erythritol / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D007553:Isotope Labeling / Q000379:methods*; D009097:Multienzyme Complexes / Q000737:chemistry / Q000378:metabolism*; D010088:Oxidoreductases / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013403:Sugar Phosphates / Q000737:chemistry / Q000378:metabolism*; D013729:Terpenes / Q000737:chemistry / Q000378:metabolism*; D014316:Tritium / Q000378:metabolism
C114232:2-C-methylerythritol 4-phosphate; D001426:Bacterial Proteins; D002250:Carbon Radioisotopes; D009097:Multienzyme Complexes; D011994:Recombinant Proteins; D013403:Sugar Phosphates; D013729:Terpenes; D014316:Tritium; D010088:Oxidoreductases; C114231:1-deoxy-D-xylulose 5-phosphate reductoisomerase; D019747:Aldose-Ketose Isomerases; D004896:Erythritol