Identification, characterization and molecular adaptation of class I redox systems for the production of hydroxylated diterpenoids

  title={Identification, characterization and molecular adaptation of class I redox systems for the production of hydroxylated diterpenoids},
  author={Christian G{\"o}rner and Patrick Schrepfer and Veronika Redai and Frank H. Wallrapp and Bernhard Loll and Wolfgang Eisenreich and Martin Haslbeck and Thomas B. Br{\"u}ck},
  journal={Microbial Cell Factories},
BackgroundDe novo production of multi-hydroxylated diterpenoids is challenging due to the lack of efficient redox systems.ResultsIn this study a new reductase/ferredoxin system from Streptomyces afghaniensis (AfR·Afx) was identified, which allowed the Escherichia coli-based production of the trihydroxylated diterpene cyclooctatin, a potent inhibitor of human lysophospholipase. This production system provides a 43-fold increase in cyclooctatin yield (15 mg/L) compared to the native producer. AfR… 
Opportunities and challenges for the sustainable production of structurally complex diterpenoids in recombinant microbial systems
Focusing mainly on the validation of successful integration of engineered biosynthetic pathways into optimized terpene producing Escherichia coli, this review shall give an insight in recent progresses regarding manipulation of mostly diterpene synthases.
From microbial upcycling to biology-oriented synthesis: combining whole-cell production and chemo-enzymatic functionalization for sustainable taxanoid delivery
A holistic bio-process based on the microbial upcycling of low-value feedstocks and leading to chemo-enzymatically derived, functionalized taxanoids was established, which is favorable under economic constraints and showed improved yields, reproducibility and transferability at any scale compared to inducible taxadiene production platforms examined in this study.
Unrivalled diversity: the many roles and reactions of bacterial cytochromes P450 in secondary metabolism.
An overview of the range of chemical transformations that P 450 enzymes can catalyse within bacterial secondary metabolism is provided to provide an important resource to aid in understanding of the potential roles of P450 enzymes within newly identified bacterial biosynthetic pathways.
Isolation and Investigation of Natural Rare Earth Metal Chelating Agents From Calothrix brevissima - A Step Towards Unraveling the Mechanisms of Metal Biosorption
Water soluble compounds that form complexes with Rare Earth Elements (REE) and other metals were isolated from Calothrix brevissima biomass with chromatographic methods for the first time, indicating that the interaction between the isolated chelators and Rare Earth Metals is stable even at high pH-values.
Strategies for terpenoid overproduction and new terpenoid discovery.
Herstellung von polyzyklischen Diterpenen
Diterpenes constitute a versatile class of natural biomolecules predominantly derived from plants, fungi and prokaryotes, which makes these compounds high value commercial targets for the chemical and pharmaceutical industry.


Molecular Characterization of a Class I P450 Electron Transfer System from Novosphingobium aromaticivorans DSM12444*
The three-dimensional structures of ArR, Arx, and CYP101D1, which form a physiological class I P450 electron transfer chain, have been resolved by x-ray crystallography and the observed structural features are consistent with the ionic strength dependence of the activity.
The first structure of a bacterial diterpene cyclase: CotB2.
The product of mutant CotB2(W288G) produced the new antibiotic compound (1R,3E,7E,11S,12S)-3,7,18-dolabellatriene, which acts specifically against multidrug-resistant Staphylococcus aureus, which opens a sustainable route for the industrial-scale production of this bioactive compound.
Structural biology of redox partner interactions in P450cam monooxygenase: a fresh look at an old system.
Identification of CYP106A2 as a Regioselective Allylic Bacterial Diterpene Hydroxylase
CYP106A2 was identified as the first reported bacterial cytochrome P450 diterpene hydroxylase and an effective whole‐cell catalyst for the selective allylic 12α‐ and 12β‐hydroxylation was applied to produce the hydroxyated products.
Characterization of the versatile monooxygenase CYP109B1 from Bacillus subtilis
CYP109B1 was found to oxidize saturated fatty acids and their methyl and ethyl esters at subterminal positions with a preference for the carbon atoms C11 and C12 counted from the carboxyl group and indole was demonstrated to inhibit fatty acid oxidation.
A novel semi-biosynthetic route for artemisinin production using engineered substrate-promiscuous P450(BM3).
This work illustrates how key P450(BM3) active site mutations enable binding of the non-native substrate amorphadiene and demonstrates high-yielding, selective transformations to dihydroartemisinic acid, the immediate precursor to the high-value antimalarial drug artemisinin.
A cDNA Clone for Taxadiene Synthase, the Diterpene Cyclase That Catalyzes the Committed Step of Taxol Biosynthesis
Sequence comparisons with monoterpene, sesquiterpenes, and diterpene cyclases of plant origin indicate a significant degree of similarity between these enzymes; the taxadiene synthase most closely resembles (46% identity, 67% similarity) abietadienes synthase, a diterPene cyclase from grand fir.
A Single Mutation at the Ferredoxin Binding Site of P450 Vdh Enables Efficient Biocatalytic Production of 25‐Hydroxyvitamin D3
Crystallographic and kinetic analyses indicate that the T107A mutation results in conformational change from an open to a closed state, thereby increasing the binding affinity with ferredoxin, and the efficient biocatalytic synthesis of 25(OH)VD3 is reported, a promising intermediate for the synthesis of various hydroxylated VD3 derivatives.