Regioselective hydroxylation of norisoprenoids by CYP109D1 from Sorangium cellulosum So ce56

Abstract

Sesquiterpenes are particularly interesting as flavorings and fragrances or as pharmaceuticals. Regio- or stereoselective functionalizations of terpenes are one of the main goals of synthetic organic chemistry, which are possible through radical reactions but are not selective enough to introduce the desired chiral alcohol function into those compounds. Cytochrome P450 monooxygenases are versatile biocatalysts and are capable of performing selective oxidations of organic molecules. We were able to demonstrate that CYP109D1 from Sorangium cellulosum So ce56 functions as a biocatalyst for the highly regioselective hydroxylation of norisoprenoids, alpha- and beta-ionone, which are important aroma compounds of floral scents. The substrates alpha- and beta-ionone were regioselectively hydroxylated to 3-hydroxy-alpha-ionone and 4-hydroxy-beta-ionone, respectively, which was confirmed by (1)H NMR and (13)C NMR. The results of docking alpha- and beta-ionone into a homology model of CYP109D1 gave a rational explanation for the regio-selectivity of the hydroxylation. Kinetic studies revealed that alpha- and beta-ionone can be hydroxylated with nearly identical V (max) and K (m) values. This is the first comprehensive investigation of the regioselective hydroxylation of norisoprenoids by CYP109D1.

Conversions of Tricyclic Antidepressants and Antipsychotics with Selected P450s fromSorangium cellulosumSo ce56

“…Therefore, we investigated 10 P450s from S. cellulosum-a bacterium typically found in soil (Shimkets et al, 2006), where it is exposed to a variety of xenobiotics-for their ability to convert tricyclic drugs. Terpenes and terpenoids as well as fatty acids have already been identified as substrates for these wild-type P450s in previous studies (Khatri et al, 2010a(Khatri et al, ,b, 2014Ly et al, 2012;Schifrin et al, 2014). Moreover, 4-methyl-3-phenyl-coumarin, a tricyclic molecule, was identified as a substrate for CYP264A1 .…”

“…For the past few years, we have been engaged in the study of novel P450s from myxobacteria, and some P450s from S. cellulosum So ce56 have shown a potential for industrial and biotechnological applications (Khatri et al, 2010a(Khatri et al, ,b, 2013(Khatri et al, , 2014Ly et al, 2012;Ringle et al, 2013). We were also able to develop an efficient E. coli-based whole-cell bioconversion system for some myxobacterial P450s .…”

Ligand characterization of CYP4B1 isoforms modified for high-level expression inEscherichia coliand HepG2 cells

“…Spectral dissociation constants were determined as described previously Khatri et al, 2010). A double beam UV/VIS spectrometer (Lambda 35, PerkinElmer, Germany) was equipped with two tandem quartz cuvettes (Hellma, Germany).…”

Efficient production of active form of vitamin D3 by microbial conversion

“…As a result, it was confirmed that the active forms of VD 3 were produced when the R. erythropolis cell was used. It has been reported that the redox partner that is capable of most efficiently supplying the electrons to P450 is not necessarily the protein with which P450 couples in the cell of the original organism [7] . This is thought to be because the electron transfer efficiency is affected drastically by the slight difference in the intracellular environment or the intracellular expression level of the gene.…”