New Sesquiterpene Oxidations with CYP260A1 and CYP264B1 from Sorangium cellulosum So ce56

Abstract

Sesquiterpenes are natural products derived from the common precursor farnesyl pyrophosphate (FPP) but are highly diverse in structure and function. Cytochrome P450 enzymes (P450s) exhibit the unique ability to introduce molecular oxygen into non-activated C-H bonds. In plant biosynthetic pathways, P450s commonly derivatize sesquiterpene hydrocarbons. However, the potential of bacterial P450s for terpene derivatization is still underinvestigated. This work compares the substrate specificities and regioselectivities of the sesquiterpene hydroxylases CYP260A1 and CYP264B1 from myxobacterium Sorangium cellulosum So ce56. Four tested substrate classes (eremophilanes, humulanes, caryophyllanes, and cedranes) were converted by both P450s. The achievable variety of oxidations is demonstrated on the model substrates (+)-nootkatone and zerumbone. Increasing the number of functionally investigated P450s, this study represents a step towards the selective derivatization of sesquiterpenes.

Improvement of a P450-Based Recombinant Escherichia coli Whole-Cell System for the Production of Oxygenated Sesquiterpene Derivatives

“…To date, several P450 enzymes have been reported to be capable of oxidizing diverse sesquiterpene substrates, such as premnaspirodiene oxygenase from the plant Hyoscyamus muticus, CYP71AV8 from the plant Cichorium intybus L., and CYP260A1 and CYP264B1 from the myxobacterium Sorangium cellulosum So ce56. Among them, CYP264B1 is a promising enzyme for the biotransformation of sesquiterpenes due to its ability to perform regioselective hydroxylation on various structural sesquiterpene classes. , Because P450s from bacteria are often not membrane associated and normally exhibit higher stabilities and activities, they are more desirable for practical application than those from plants . Thus, CYP264B1 is an attractive candidate for the oxidation of sesquiterpene compounds.…”

Structural characterization of CYP260A1 from Sorangium cellulosum to investigate the 1α‐hydroxylation of a mineralocorticoid

“…Among them, CYP260A1 is one of the novel P450s, which we have recently shown to perform a selective 1α‐hydroxylation of C‐19 steroids . In addition, the oxidation of sesquiterpenes and conversion of drugs by CYP260A1 was also demonstrated. Although its autologous redox partners Fdx2/FdRB and Fdx8/FdRB were identified as efficient for sesquiterpene oxidation , the bovine redox partners adrenodoxin (Adx 4–108 ) and adrenodoxin reductase (AdR) were shown to be efficient surrogate redox partners , and they were employed for the establishment of an Escherichia coli ‐based whole‐cell system .…”

Characterization and structure‐guided engineering of the novel versatile terpene monooxygenase CYP109Q5 from Chondromyces apiculatus DSM436

“…There are only few examples for biocatalysts with activity towards the tested sesquiterpenes such as α‐humulene, β‐bisabolene or premnaspirodiene, which makes the enzyme CYP109Q5 particularly interesting. The previous examples for the production of 5‐ or 8‐hydroxy‐α‐humulene were extended by the biocatalyst identified in this work (Yu et al ., ; Schifrin et al ., ). For the first time, oxyfunctionalization of β‐bisabolene was enabled by a heterologously expressed bacterial P450.…”

“…For the identification of novel terpene hydroxylases, we therefore searched for organisms that possess large numbers of potentially interesting P450s for the oxyfunctionalization of terpenes. Studies on the myxobacterium S. cellulosum So ce56 have already identified and characterized P450s, able to efficiently hydroxylate norisoprenoids and sesquiterpenes (Ly et al ., ; Schifrin et al ., ; Litzenburger and Bernhardt, ). Myxobacteria feature the largest bacterial genomes known today and a versatile secondary metabolism, making them particularly interesting for biotechnological applications (Schneiker et al ., ; Diez et al ., ).…”