A5070268234- Microbial Natural Products and Biosynthesis
- Enzyme Structure and Function
- Biochemical and Structural Characterization
- Bacterial Genetics and Biotechnology
- RNA and protein synthesis mechanisms
- Biochemical and Molecular Research
- Chemical Synthesis and Analysis
- Amino Acid Enzymes and Metabolism
- Microbial Metabolic Engineering and Bioproduction
- DNA Repair Mechanisms
- Photosynthetic Processes and Mechanisms
- Marine Sponges and Natural Products
- RNA modifications and cancer
- DNA and Nucleic Acid Chemistry
- Metalloenzymes and iron-sulfur proteins
- Carbohydrate Chemistry and Synthesis
- Genomics and Phylogenetic Studies
- Metal-Catalyzed Oxygenation Mechanisms
- Plant biochemistry and biosynthesis
- Porphyrin Metabolism and Disorders
- Cancer therapeutics and mechanisms
- Chemical synthesis and alkaloids
- Microbial Metabolism and Applications
- Metabolism and Genetic Disorders
- Drug Transport and Resistance Mechanisms
University of Florida
2015-2024
Gainesville Obstetrics & Gynecology
2015-2018
Boston College
1995-2009
Harvard University
1959-2008
University of Wisconsin–Madison
2007
Philipps University of Marburg
2002
Harvard University Press
2000
Kyoto University
1959
Kyoto City University of Arts
1959
City College
1959
Certain commensal Escherichia coli contain the clb biosynthetic gene cluster that codes for small molecule prodrugs known as precolibactins. Precolibactins are converted to colibactins by N-deacylation; latter postulated be genotoxic and contribute colorectal cancer formation. Though advances toward elucidating (pre)colibactin biosynthesis have been made, functions mechanisms of several products remain poorly understood. Here we report 2.1 Å X-ray structure molecular function ClbS, a product...
Nonribosomal peptide synthetases are large, complex multidomain enzymes responsible for the biosynthesis of a wide range peptidic natural products. Inherent to synthetase chemistry is thioester templated mechanism that relies on protein/protein interactions and interdomain dynamics. Several questions related structure remain be addressed, including incorporation accessory domains intermodule interactions. The inclusion nonproteinogenic d-amino acids into frameworks common important...
Dolastatin 10, a potent tubulin-targeting marine anticancer natural product, provided the basis for development of six FDA-approved antibody–drug conjugates. Through screening cyanobacterial Caldora penicillata environmental DNA libraries and metagenome sequencing, we identified its biosynthetic gene cluster. Functional prediction 10 enzymes encoded in 39 kb cluster supports dolastatin biosynthesis. The nonheme diiron monooxygenase DolJ was biochemically characterized to mediate terminal...
Fidelity and efficiency. Nonribosomal peptide synthetases, which contain domains for the activation (A), thiolation (T), condensation (C) of amino acids (AA), are high-efficiency, high-fidelity assembly lines synthesizing natural products. Errors in a single step can have serious consequences product formation. Type II thioesterases (TEII) might play critical role ensuring efficiency accuracy these systems, essential features any attempt to engineer NRPSs produce novel
The C-terminal thioesterase domain of the nonribosomal peptide synthetase producing lipopetide surfactin (Srf TE) retains autonomous ability to generate cyclic peptidolactone skeleton when provided with a soluble β-hydroxy-butyryl-heptapeptidyl thioester substrate. Utilizing recently solved crystal structure [Bruner, S. D., et al. (2002) Structure 10, 301−310], active-site nucleophile, Ser80, was changed Cys, and other members catalytic triad, Asp107 His207, were Ala, resulting mutants...
The PacC transcription factor is an important component of the fungal ambient pH-responsive regulatory system. Loss pacC in insect pathogenic fungus Beauveria bassiana resulted alkaline pH-dependent decrease growth and increased susceptibility to osmotic (salt, sorbitol) stress SDS. Extreme Congo Red was noted irrespective pH, ΔBbpacC conidia showed subtle increases UV susceptibility. ΔBbPacC mutant a reduced ability acidify media during due failure produce oxalic acid. also did not...
Continuous directed evolution of enzymes and other proteins in microbial hosts is capable outperforming classical by executing hypermutation selection concurrently vivo, at scale, with minimal manual input. Provided that a target enzyme's activity can be coupled to growth the host cells, improved simply selecting for growth. Like all evolution, continuous version requires no prior mechanistic knowledge target. thus powerful way modify plant or non-plant use metabolic research engineering....
Developing potent and selective protease inhibitors remains a grueling, iterative, often unsuccessful endeavor. Although macromolecular can achieve single-enzyme specificity, platforms used for inhibitor discovery are optimized high-affinity binders, requiring extensive downstream biochemical characterization to isolate rare inhibitors. Here, we developed the High-throughput Activity Reprogramming of Proteases (HARP) platform, HARP is yeast-based functional screen that isolates...
β-Amino acids are widely used building blocks in both natural and synthetic compounds. Aromatic β-amino can be biosynthesized directly from proteinogenic α-amino by the action of MIO (4-methylideneimidazole-5-one)-based aminomutase enzymes. The uncommon cofactor plays a role ammonia lyases 2,3-aminomutases; however, precise mechanism has not been resolved. Here we provide evidence that electrophilic uses covalent catalysis through substrate amine to direct elimination subsequent readdition...
The SgcC4 l-tyrosine 2,3-aminomutase (SgTAM) catalyzes the formation of (S)-β-tyrosine in biosynthetic pathway enediyne antitumor antibiotic C-1027. SgTAM is homologous to histidine ammonia lyase family enzymes whose activity dependent on methylideneimidazole-5-one (MIO) cofactor. Unlike enzymes, additional chemical transformations resulting an overall stereospecific 1,2-amino shift substrate generate (S)-β-tyrosine. Previously, we provided kinetic, spectroscopic, and mutagenesis data...
Bacteria belonging to the order Actinomycetales have proven be an important source of biologically active and often therapeutically useful natural products. The characterization orphan biosynthetic gene clusters is emerging valuable approach discovery novel small molecules. Analysis recently sequenced genome thermophilic actinomycete Thermobifida fusca revealed nonribosomal peptide cluster coding for unknown siderophore product. T. a model organism study thermostable cellulases major...