Characterization of the Gene Cluster CYP264B1‐geoA from Sorangium cellulosum So ce56: Biosynthesis of (+)‐Eremophilene and Its Hydroxylation

Abstract

Terpenoids can be found in almost all forms of life; however, the biosynthesis of bacterial terpenoids has not been intensively studied. This study reports the identification and functional characterization of the gene cluster CYP264B1-geoA from Sorangium cellulosum So ce56. Expression of the enzymes and synthesis of their products for NMR analysis and X-ray diffraction were carried out by employing an Escherichia coli whole-cell conversion system that provides the geoA substrate farnesyl pyrophosphate through simultaneous overexpression of the mevalonate pathway genes. The geoA product was identified as a novel sesquiterpene, and assigned NMR signals unambiguously proved that geoA is an (+)-eremophilene synthase. The very tight binding of (+)-eremophilene (∼0.40 μM), which is also available in S. cellulosum So ce56, and its oxidation by CYP264B1 suggest that the CYP264B1-geoA gene cluster is required for the biosynthesis of (+)-eremophilene derivatives.

Mechanistic Characterisation of Two Sesquiterpene Cyclases from the Plant Pathogenic Fungus Fusarium fujikuroi

“…GC/MS analysis ( Figure S6 A) suggested the structure of valencene or eremo-philene,but it was not possible to clearly assign one of these structures.Alarge-scale incubation of 50 mg FPP yielded 4.6 mg (19 %) of the pure sesquiterpene with 1 H-and 13 C-NMR data matching those of eremophilene (2;T able S1). [21] Thep roposed biosynthesis of 2 (Scheme 3) starts with a1 ,10-cyclization of FPP to the (E,E)-germacradienyl cation J,f ollowed by deprotonation to germacrene A (K). [21] Thep roposed biosynthesis of 2 (Scheme 3) starts with a1 ,10-cyclization of FPP to the (E,E)-germacradienyl cation J,f ollowed by deprotonation to germacrene A (K).…”

CYP267A1 and CYP267B1 from Sorangium cellulosum So ce56 are Highly Versatile Drug Metabolizers

“…During our recent investigations on P450 enzymes from the myxobacterium Sorangium cellulosum So ce56, several interesting enzymes displaying new properties and substrate specificities have been discovered, which lead us to investigate the potential of these P450 enzymes for an application as drug metabolizing biocatalysts Schifrin et al, 2015;Litzenburger and Bernhardt, 2016). Therefore, we used bioinformatics analysis to identify myxobacterial P450 enzymes that are closely related to drug metabolizing P450 enzymes.…”

Structure–function analysis for the hydroxylation of Δ4 C21‐steroids by the myxobacterial CYP260B1

“…We found that the strain Sorangium cellulosum So ce56 contains 21 P450s, 8 ferredoxins, and 2 ferredoxin reductases . However, only seven of the P450s, belonging to families CYP109 (CYP109C1, CYP109C2, and CYP109D1) , CYP264 (CYP264A1 and CYP264B1) , and CYP267 (CYP267A1 and CYP267B1) , were well characterized either for potential physiological or biotechnological applications. They are involved in the hydroxylation of fatty acids, terpenes, and terpenoids, as well as of drugs and drug derivatives .…”