A5037827364- Microbial Natural Products and Biosynthesis
- Biochemical and Structural Characterization
- Chemical Synthesis and Analysis
- Chemical Reactions and Mechanisms
- Fluorine in Organic Chemistry
- Radical Photochemical Reactions
- History and advancements in chemistry
- Carbon dioxide utilization in catalysis
- Enzyme Catalysis and Immobilization
- Antimicrobial Peptides and Activities
- Peptidase Inhibition and Analysis
- Pharmacological Effects of Natural Compounds
- Plant biochemistry and biosynthesis
- CO2 Reduction Techniques and Catalysts
- Enzyme Structure and Function
- Molecular spectroscopy and chirality
- Photosynthetic Processes and Mechanisms
- Genomics and Phylogenetic Studies
- RNA and protein synthesis mechanisms
- Phytochemistry and Biological Activities
- Biochemical and Molecular Research
- Gout, Hyperuricemia, Uric Acid
- Chemical Reactions and Isotopes
- Cyclopropane Reaction Mechanisms
- Carbohydrate Chemistry and Synthesis
Leibniz-Institut für Naturstoff-Forschung und Infektionsbiologie e. V. - Hans-Knöll-Institut (HKI)
2018-2024
University of Bayreuth
2023-2024
Naturstoff-Technik (Germany)
2022
ETH Zurich
2010-2020
Norwich Research Park
2015-2018
John Innes Centre
2015-2018
Nonribosomal peptide synthetases (NRPSs) protect microorganisms from environmental threats by producing diverse siderophores, antibiotics, and other natural products. Their modular molecular structure is also attractive the standpoint of biosynthetic engineering. Here we evaluate a methodology for swapping module specificities these mega-enzymes that takes advantage flavodoxin-like subdomains involved in substrate recognition. Nine encoding were transplanted into Phe-specific GrsA initiation...
Abstract Nonribosomal peptide synthetases (NRPSs) are multifunctional enzymes that produce a wide array of bioactive peptides. Here we show single tryptophan‐to‐serine mutation in phenylalanine‐specific NRPS adenylation domains enables the efficient activation non‐natural aromatic amino acids functionalized with azide and alkyne groups. The resulting 10 5 ‐fold switch substrate specificity was achieved without appreciable loss catalytic efficiency. Moreover, effective communication modified...
The secoiridoids are the main class of specialized metabolites present in olive (Olea europaea L.) fruit. In particular, secoiridoid oleuropein strongly influences oil quality because its bitterness, which is a desirable trait. addition, possesses wide range pharmacological properties, including antioxidant, anti-inflammatory, and anti-cancer activities. accordance, obtaining high varieties goal molecular breeding programs. Here we use transcriptomic approach to identify candidate genes...
Temperature influences the reaction kinetics and evolvability of all enzymes. To understand how evolution shapes thermodynamic drivers catalysis, we optimized modest activity a computationally designed enzyme for an elementary proton-transfer by nearly 4 orders magnitude over 9 rounds mutagenesis screening. As theorized primordial enzymes, catalytic effects original design were almost entirely enthalpic in origin, as rate enhancements achieved laboratory evolution. However, large reductions...
Abstract Bacteria of the Burkholderia pseudomallei (BP) group pose a global health threat, causing infectious diseases melioidosis, common cause pneumonia and sepsis, glanders, contagious zoonosis. A trait BP bacteria is conserved gene cluster coding for biosynthesis polyketides (malleicyprols) with reactive cyclopropanol unit that critical virulence. Enzymes building this warhead represent ideal targets antivirulence strategies but biochemical basis formation unknown. Here we describe...
Engineering of biosynthetic enzymes is increasingly employed to synthesize structural analogues antibiotics. Of special interest are nonribosomal peptide synthetases (NRPSs) responsible for the production important antimicrobial peptides. Here, directed evolution an adenylation domain a Pro-specific NRPS module completely switched substrate specificity non-standard amino acid piperazic (Piz) bearing labile N-N bond. This success was achieved by UPLC-MS/MS-based screening small, rationally...
Cyclopropanol rings are highly reactive and may function as molecular "warheads" that affect natural product bioactivity. Yet, knowledge on their biosynthesis is limited. Using gene cluster analyses, isotope labeling, in vitro enzyme assays, we shed first light the of cyclopropanol-substituted amino acid cleonine, a residue antimicrobial depsipeptide valgamicin C cytotoxic glycopeptide cleomycin A2. We decipher biosynthetic origin show cleonine cyclopropanol ring derived from...
Nonribosomal peptide synthetases (NRPSs) are sophisticated molecular machines that biosynthesize drugs. In attempts to generate new bioactive compounds, some parts of NRPSs have been successfully manipulated, but especially the influence condensation (C-)domains on substrate specificity remains enigmatic and poorly controlled. To understand C-domains preference, we extensively evaluated formation C-domain mutants in a bimodular NRPS system. Thus, identified three key mutations govern...
The site-specific conjugation of polymers to multiple engineered cysteine residues a prolyl endopeptidase leads its stabilization in the gastrointestinal tract rats, without compromising activity relative native enzyme. importance polymer attachment sites is investigated, as well significance structure. As service our authors and readers, this journal provides supporting information supplied by authors. Such materials are peer reviewed may be re-organized for online delivery, but not...
Nonribosomal peptide synthetases (NRPSs) are a vast source of valuable natural products, and re-engineering them is an attractive path toward structurally diversified active compounds. NRPS engineering often requires heterologous expression, which hindered by the enormous size proteins. Protein splitting docking domain insertion have been proposed as strategy to overcome this limitation. Here, we applied gramicidin S NRPS: Despite better production split proteins, almost ceased. However,...
Adenylation enzymes are engineering targets in ribosomal and nonribosomal peptide synthesis. Through multiplexed LC-MS/MS measurement of hydroxamates, the HAMA assay records specificity profiles these a snap.
A snapshot of evolution <italic>in flagrante</italic> shows that recombination within and between biosynthetic genes leads to diversification nonribosomal peptides.
Engineering of nonribosomal peptide synthetases (NRPSs) has faced numerous obstacles despite being an attractive path toward bioactive molecules. Specificity filters in the assembly line determine engineering success, but relative contribution adenylation (A) and condensation (C) domains is under debate. In engineered, bimodular NRPS sdV-GrsA/GrsB1, first module a subdomain-swapped chimera showing substrate promiscuity. On sdV-GrsA evolved mutants, we have employed kinetic modeling to...
Pathogenic bacteria of the Burkholderia pseudomallei group cause severe infectious diseases such as glanders and melioidosis. Malleicyprols were identified important bacterial virulence factors, yet biosynthetic origin their cyclopropanol warhead has remained enigmatic. By a combination mutational analysis metabolomics we found that sulfonium acids, dimethylsulfoniumpropionate (DMSP) gonyol, known osmolytes crucial components in global organosulfur cycle, are key intermediates en route to...