Regioselective biooxidation of (+)-valencene by recombinant E. coli expressing CYP109B1 from Bacillus subtilis in a two-liquid-phase system

Abstract

Background: (+)-Nootkatone (4) is a high added-value compound found in grapefruit juice. Allylic oxidation of the sesquiterpene (+)-valencene (1) provides an attractive route to this sought-after flavoring. So far, chemical methods to produce (+)-nootkatone (4) from (+)-valencene (1) involve unsafe toxic compounds, whereas several biotechnological approaches applied yield large amounts of undesirable byproducts. In the present work 125 cytochrome P450 enzymes from bacteria were tested for regioselective oxidation of (+)-valencene (1) at allylic C2-position to produce (+)-nootkatone (4) via cis-(2) or trans-nootkatol (3). The P450 activity was supported by the coexpression of putidaredoxin reductase (PdR) and putidaredoxin (Pdx) from Pseudomonas putida in Escherichia coli.

Bioprocess Engineering for Microbial Synthesis and Conversion of Isoprenoids

“…Usually a number of organic solvents with appropriate log P Oct values are compiled and experiments conducted to determine biocompatibility, partition coefficients for substrate, and/or product and biodegradability [6,22,37,56,80]. To test for biocompatibility an activity profile can be created by plotting an organic solvent log P Oct against biocatalyst activity.…”

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

“…[16][17][18][19][20] Volatile mono-(two isoprene subunits) and sesquiterpenoids (three isoprene subunits) are postulated to serve as means of chemical communication in animals, plants, insects, and microorganisms. [21][22][23] For industrial production, plant terpenoids are of particularly high value, [24] because of their wide application as flavors and fragrances such as the grapefruit odorant (+)-nootkatone (Scheme 2, 1 b), [25][26][27][28][29][30] as highly potent pharmaceuticals like the taxanes in tumor therapy, [31][32] and the anti-malarial drug artemisinin. [33] Cytochrome P450s are able to perform terpenoid hydroxylations [29,30] in a way that is superior to synthetic chemistry concerning selectivity and efficiency.…”

“…[21][22][23] For industrial production, plant terpenoids are of particularly high value, [24] because of their wide application as flavors and fragrances such as the grapefruit odorant (+)-nootkatone (Scheme 2, 1 b), [25][26][27][28][29][30] as highly potent pharmaceuticals like the taxanes in tumor therapy, [31][32] and the anti-malarial drug artemisinin. [33] Cytochrome P450s are able to perform terpenoid hydroxylations [29,30] in a way that is superior to synthetic chemistry concerning selectivity and efficiency. [34][35][36] Escherichia coli and Bacillus megaterium have proven to be valuable hosts for in vivo cytochrome P450 bioconversions of terpenoids.…”

One-Pot Synthesis of (+)-Nootkatone via Dark Singlet Oxygenation of Valencene: The Triple Role of the Amphiphilic Molybdate Catalyst

“…They obtained (+)-nootkatone with a yield of 40%. On the other hand, various biocatalysts, such as G. pentaphyllum cultures, green algae Chlorella species, fungi Bothryosphaeria dothidea, the lyophilisate of edible mushroom Pleurotus sapidus, and several bacterial cytochrome P450 enzymes have also been studied for this transformation [27][28][29][30][31][32][33][34][35]. However, the costly culture conditions, the low conversion rate and yield, the inhibition of enzymes by products, and the presence of various by-products still hamper the industrial preparation of (+)-nootkatone via biocatalysts.…”