A5069672847- Plant biochemistry and biosynthesis
- Microbial Natural Products and Biosynthesis
- Natural product bioactivities and synthesis
- Sesquiterpenes and Asteraceae Studies
- Enzyme Structure and Function
- Fungal Biology and Applications
- Biocrusts and Microbial Ecology
- Lipid metabolism and biosynthesis
- Pharmacological Effects of Natural Compounds
- Plant Pathogens and Fungal Diseases
- Phytochemical compounds biological activities
- Microbial Metabolism and Applications
- Microbial Metabolic Engineering and Bioproduction
- Algal biology and biofuel production
- Mycotoxins in Agriculture and Food
- Phytochemistry and Biological Activities
- Biochemical and Molecular Research
- Heat shock proteins research
- Polyamine Metabolism and Applications
- Microbial Applications in Construction Materials
- Synthetic Organic Chemistry Methods
- Neurobiology and Insect Physiology Research
- Biochemical and biochemical processes
- Protein Structure and Dynamics
- X-ray Diffraction in Crystallography
University of Florida
2024
University of Bonn
2017-2023
The Herbert Wertheim UF Scripps Institute for Biomedical Innovation & Technology
2023
Czech Academy of Sciences, Institute of Organic Chemistry and Biochemistry
2020
All known triterpenes are generated by triterpene synthases (TrTSs) from squalene or oxidosqualene1. This approach is fundamentally different the biosynthesis of short-chain (C10-C25) terpenes that formed polyisoprenyl diphosphates2-4. In this study, two fungal chimeric class I TrTSs, Talaromyces verruculosus talaropentaene synthase (TvTS) and Macrophomina phaseolina macrophomene (MpMS), were characterized. Both enzymes use dimethylallyl diphosphate isopentenyl hexaprenyl as substrates,...
Two diterpene synthases from Allokutzneria albata were studied for their products, resulting in the identification of new compound bonnadiene first enzyme. Although phylogenetically unrelated to fungal phomopsene synthase, second enzyme produced a mixture and biosynthetically linked compound, allokutznerene, as well spiroviolene. Both enzymes subjected in-depth mechanistic studies involving isotopic labelling experiments, metal-cofactor variation, site-directed mutagenesis. Oxidation...
Two bacterial diterpene synthases from the actinomycete Allokutzneria albata were investigated, resulting in identification of structurally unprecedented compound spiroalbatene first and cembrene A second enzyme. Both enzymes thoroughly investigated terms their mechanisms by isotope labeling experiments, site-directed mutagenesis, variation metal cofactors pH value. For synthase, pH- Mn2+ -dependent formation side product thunbergol was observed, which is biosynthetically linked to spiroalbatene.
A terpene synthase from the marine bacterium Streptomyces xinghaiensis has been characterised, including a full structure elucidation of its products various substrates and an in-depth investigation enzyme mechanism by isotope labelling experiments, metal cofactor variations, mutation experiments. The results revealed interesting dependency Mn2+ catalysis on presence Asp-217, residue that is occupied highly conserved Glu in most other bacterial synthases.
Based on a terpenoid overproduction platform in yeast for genome mining, chimeric diterpene synthase from the endophytic fungus Colletotrichum gloeosporioides ES026 was characterized as (5R,12R,14S)-dolasta-1(15),8-diene synthase. The absolute configuration independently verified through use of enantioselectively deuterated terpene precursors, which unequivocally established predicted C1-III-IV cyclization mode this first clade II-D enzyme. Extensive isotopic labeling experiments and...
The substrate flexibility of eight purified sesquiterpene cyclases was evaluated using six new heteroatom-modified farnesyl pyrophosphates, and the formation macrocyclic tricyclic sesquiterpenoids is described. GC-O analysis revealed that tetrahydrofuran exhibits an ethereal, peppery, camphor-like olfactoric scent.
Abstract A diterpene synthase from Saccharopolyspora spinosa was found to convert geranylgeranyl diphosphate into the new natural products spinodiene and B, accompanied by 2,7,18‐dolabellatriene. The structures formation mechanism of enzyme were investigated extensive isotopic labelling experiments, which revealed an unusual branched isomerisation towards neutral intermediate Diels–Alder reaction used main product with its rare conjugated diene moiety formal sesterterpene alcohols.
Abstract Zwei Diterpensynthasen aus Allokutzneria albata wurden auf ihre Produkte untersucht. Das Produkt des ersten Enzyms wurde als die neue Verbindung Bonnadien identifiziert. Obwohl das zweite Enzym phylogenetisch nicht mit der pilzlichen Phomopsen‐Synthase in steht, produzierte es eine Mischung Phomopsen und einer biosynthetisch verwandten neuen Verbindung, Allokutzneren, sowie Spiroviolen. Beide Enzyme im Detail durch Isotopenmarkierungsexperimente, Variation Metallcofaktors...
Fusarium culmorum is one of the most important fungal plant pathogens that causes diseases on a wide diversity cereal and non-cereal crops. We report herein for first time genome sequence F. strain PV its associated secondary metabolome plays role in interaction with other microorganisms contributes to pathogenicity plants. The revealed presence two terpene synthases, trichodiene longiborneol synthase, which generate an array volatile terpenes. Furthermore, we identified gene clusters,...
Two bacterial diterpene synthases (DTSs) from Chryseobacterium were characterised. The first enzyme yielded the new compound chryseodiene that closely resembles known fusicoccane diterpenes fungi, but its experimentally and computationally studied cyclisation mechanism is fundamentally different to of fusicoccadiene synthase. second produced wanjudiene, a hydrocarbon with skeleton, besides traces enantiomer bonnadiene was recently discovered Allokutzneria albata.
Methylated analogues of isopentenyl diphosphate were synthesised and enzymatically incorporated into methylated terpenes. A detailed stereochemical analysis the obtained products is presented. The terpene precursors also used in conjunction with various isotopic labellings to gain insights mechanisms their enzymatic formation.
Abstract Zwei bakterielle Diterpensynthasen aus dem Actinomyceten Allokutzneria albata wurden untersucht. Dies führte zur Identifizierung des strukturell neuartigen Produkts Spiroalbaten ersten und Cembren A zweiten Enzyms. Beide Enzyme durch Isotopenmarkierungsexperimente, ortsgerichtete Mutagenese, Variation Metallcofaktors pH‐Wertes auf ihre Mechanismen Für die Spiroalbaten‐Synthase wurde eine pH‐ Mn 2+ ‐abhängige Bildung Nebenproduktes Thunbergol beobachtet, das biosynthetisch mit verwandt ist.
Fusicoccane (FC)-type diterpenes, featuring a common 5–8–5 tricyclic skeleton, possess diverse biological functions. Currently, only FC-type diterpene synthases (DTSs) for 3 of the 16 possible stereochemically distinct subtypes FC skeleton have been discovered. Herein, (2si,6si,10re,14si)-FC-type DTS from Myrothecium gramineum is reported, which produces alcohol myrothec-15(17)-en-7-ol. On basis homology modelings, I189F variant was obtained by site-directed mutagenesis that prevents...
Dimethylsulfoniopropionate (DMSP) catabolism of marine bacteria plays an important role in and global ecology. The genome Ruegeria pomeroyi DSS-3, a model organism from the Roseobacter group, harbours no less than three genes for different DMSP lyases (DddW, DddP DddQ) that catalyse degradation to dimethyl sulfide (DMS) acrylate. Despite their apparent similar function these enzymes show significant overall sequence identity. In this work DddQ DddW R. homolog Phaeobacter inhibens DSM 17395...
Abstract The skeletons of some classes terpenoids are unusual in that they contain a larger number Me groups (or their biosynthetic equivalents such as olefinic methylene groups, hydroxymethyl aldehydes, or carboxylic acids and derivatives) than provided by oligoprenyl diphosphate precursor. This is sometimes the result an oxidative ring‐opening reaction at terpene‐cyclase‐derived molecule containing regular group equivalents, observed for picrotoxan sesquiterpenes. In this study...
Confinement of an enzyme's active site is critical to the efficiency chemical reactions and has been recognized as important tool for catalysis accelerate by proximity via specific interactions with substrate control course reaction. Despite these benefits, a strongly confined inherently limited compounds that resemble native substrate, only small deviations tolerated. To unlock potential modifying terpene scaffold building blocks interest branched isoprene/terpene motifs, present study...
Abstract Eine Terpensynthase aus dem marinen Bakterium Streptomyces xinghaiensis wurde charakterisiert, einschließlich einer vollen Strukturaufklärung ihrer Produkte verschiedenen Substraten. Weiterhin wurden detaillierte Untersuchungen des Enzymmechanismus durch Isotopenmarkierungsexperimente, Variationen Metallcofaktors und Mutageneseexperimente durchgeführt. Die Daten zeigten eine Abhängigkeit der Mn 2+ ‐katalysierten Reaktion von Gegenwart Asp‐217, einem Rest, in anderen bakteriellen...
Abstract Eine Diterpensynthase aus Saccharopolyspora spinosa konvertierte Geranylgeranyldiphosphat in die neuen Naturstoffe Spinodien A und B, begleitet von 2,7,18‐Dolabellatrien. Die Strukturen Bildungsmechanismen der Enzymprodukte wurden mithilfe Isotopenmarkierungsexperimenten studiert, wodurch ein ungewöhnlicher verzweigter Isomerisierungsmechanismus zu dem neutralen Intermediat 2,7,18‐Dolabellatrien aufgedeckt wurde. Durch eine Diels‐Alder‐Reaktion wurde das Diterpen‐Hauptprodukt mit...
Abstract Basierend auf einer Plattform zur Überproduktion von Terpenoiden in Hefe für “Genome‐Mining” wurde eine chimäre Diterpensynthase aus dem endophytischen Pilz Colletotrichum gloeosporioides ES026 als (5 R ,12 ,14 S )‐Dolasta‐1(15),8‐dien‐Synthase charakterisiert. Die absolute Konfiguration unabhängig durch die Verwendung enantioselektiv deuterierter Terpenvorstufen bestimmt, wodurch der vorhergesagte C1‐III‐IV‐Cyclisierungsmodus dieses ersten charakterisierten Enzyms Klade II‐D...
Actinobacteria, the bacterial phylum most renowned for natural product discovery, has been established as a valuable source drug discovery and biotechnology but is underrepresented within accessible genome strain collections. Herein, we introduce Natural Products Discovery Center (NPDC), featuring 122,449 strains assembled over eight decades, genomes of first 8490 NPDC (7142 Actinobacteria), online Portal making both publicly available. A comparative survey RefSeq Actinobacteria highlights...
Abstract Die Substratflexibilität von acht rekombinant erzeugten Sesquiterpencyclasen wurde unter Anwendung sechs neuen Heteroatom‐modifizierten Farnesylpyrophosphaten untersucht. Als Ergebnis werden neue makro‐ oder tricyclische, Heteroatom‐modifizierte Sesquiterpene beschrieben. GC‐O‐Analyse zeigte, dass das tricyclische Tetrahydrofuran etherische, pfeffrige und campherartige Geruchsnoten aufweist.
Abstract Zwei bakterielle Diterpensynthasen (DTS) aus Chryseobacterium wurden charakterisiert. Das erste Enzym ergab die neue Verbindung Chryseodien, nahe mit den Fusicoccan‐Diterpenen Pilzen verwandt ist, allerdings ist ihr experimentell und computerchemisch studierter Cyclisierungsmechanismus fundamental unterschiedlich zum Mechanismus der Fusicoccadien‐Synthase. zweite produzierte Wanjudien, einen Diterpenkohlenwasserstoff neuem Skelett, neben Spuren des Enantiomers von Bonnadien, das...