Burkholderia cenocepacia: a new biocatalyst for efficient bioreduction of ezetimibe intermediate.
Ezetimibe is a selective acyl-coenzyme A: cholesterol acyltransferase (ACAT) inhibitor used in hypercholesterolemia. Synthesis of ezetimibe requires enantiopure 3-[5-(4-fluorophenyl)-5(S)-hydroxypentanoyl]-4(S)-4-phenyl-1,3-oxazolidin-2-one (FOP alcohol) as a crucial intermediate which is produced by reduction of the corresponding prochiral ketone (FOP dione). A new biocatalyst from acclimatized soil was screened for bioreduction of the above prochiral ketone. The microorganism was identified by 16S mRNA sequencing, as Burkholderia cenocepacia. Various physicochemical conditions were optimized to increase cellmass and enzyme activity. The overall increase in cellmass concentration and enzyme activity was 2.06 and 1.85-fold, respectively. Various reaction conditions, for example pH, temperature, agitation, and cellmass concentration, were optimized for maximum product yield (chiral alcohol) with excellent enantioselectivity. Best reduction was achieved in phosphate buffer (50 mM, pH 8.0) at 40 degrees C (200 rpm) and the yield of enantiopure alcohol from the corresponding prochiral ketone was 54%. This biocatalyst was also used for the reduction of various other prochiral ketones.
Singh
Amit
A
Department of Pharmaceutical Technology (Biotechnology), National Institute of Pharmaceutical Education and Research, Mohali, Punjab, India.
Basit
Abdul
A
Banerjee
Uttam C
UC
D000924:Anticholesteremic Agents / Q000378:metabolism*; D001384:Azetidines / Q000378:metabolism*; D053898:Biosynthetic Pathways; D001711:Biotransformation; D019117:Burkholderia / Q000145:classification / Q000254:growth & development* / Q000302:isolation & purification / Q000378:metabolism*; D016000:Cluster Analysis; D003067:Coenzymes / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D000069438:Ezetimibe; D006863:Hydrogen-Ion Concentration; D008670:Metals / Q000378:metabolism; D008954:Models, Biological; D015394:Molecular Structure; D010084:Oxidation-Reduction; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D013696:Temperature
D000924:Anticholesteremic Agents; D001384:Azetidines; D003067:Coenzymes; D003470:Culture Media; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D008670:Metals; D012336:RNA, Ribosomal, 16S; D000069438:Ezetimibe