A5045344351- Microbial Natural Products and Biosynthesis
- Enzyme Production and Characterization
- Enzyme Structure and Function
- Carbohydrate Chemistry and Synthesis
- Enzyme Catalysis and Immobilization
- Genomics and Phylogenetic Studies
- Microbial Metabolic Engineering and Bioproduction
- Plant biochemistry and biosynthesis
- Synthetic Organic Chemistry Methods
- Glycosylation and Glycoproteins Research
- Chemical Synthesis and Analysis
- Biofuel production and bioconversion
- Biochemical and Molecular Research
- Fungal Biology and Applications
- Peptidase Inhibition and Analysis
- Phytochemical compounds biological activities
- Bacterial Genetics and Biotechnology
- Fungal and yeast genetics research
- Microbial Metabolism and Applications
- Force Microscopy Techniques and Applications
- Cellular Mechanics and Interactions
- Microbial Metabolites in Food Biotechnology
- Bacteriophages and microbial interactions
- Plant Pathogens and Fungal Diseases
- Protein Structure and Dynamics
The University of Texas at Austin
2016-2025
Brown University
2008-2010
Stanford University
1998-2010
University of California, San Francisco
2001-2008
While engineering modular polyketide synthases (PKSs) using the recently updated module boundary has yielded libraries of triketide-pentaketides, this strategy not yet been applied to combinatorial biosynthesis macrolactones or macrolide antibiotics. We developed a 2-plasmid system for construction and expression PKSs employed it obtain refactored pikromycin synthase in E. coli that produces 85 mg narbonolide per liter culture. The replacement, insertion, deletion, mutagenesis modules...
ADVERTISEMENT RETURN TO ISSUEPREVCommunicationNEXTHigh-Resolution Macromolecular NMR Spectroscopy Inside Living CellsZach Serber, Adrian T. Keatinge-Clay, Richard Ledwidge, Alexander E. Kelly, Susan M. Miller, and Volker DötschView Author Information Graduate Group in Biophysics Department of Pharmaceutical Chemistry Cellular Molecular Pharmacology University California San Francisco Francisco, 94143 Cite this: J. Am. Chem. Soc. 2001, 123, 10, 2446–2447Publication Date (Web):February 15,...
Efficient bacterial genetic engineering approaches with broad‐host applicability are rare. We combine two systems, mobile group II introns (‘targetrons’) and Cre/ lox , which function efficiently in many different organisms, into a versatile platform we call GETR (Genome Editing via Targetrons Recombinases). The deliver sites to specific genomic loci, enabling manipulations. Efficiency is enhanced by adding flexibility the RNA hairpins formed sites. use system for insertions, deletions,...
Modular redefinition: A long-standing paradigm in modular polyketide synthase enzymology, namely the definition of a module, has been challenged by Abe and co-workers their recent study. With this new understanding emerged renewed hope for engineering these assembly lines to produce materials medicines.
Abstract To harness the synthetic power of modular polyketide synthases (PKSs), many aspects their biochemistry must be elucidated. A robust platform to study these megadalton assembly lines has not yet been described. Here, we in vitro reconstitute venemycin PKS, a short line that generates an aromatic product. Incubating its polypeptides, VemG and VemH, with 3,5-dihydroxybenzoic acid, ATP, malonate, coenzyme A, malonyl-CoA ligase MatB, production can monitored by HPLC NMR. Multi-milligram...
The modular nature of polyketide assembly lines and the significance their products make them prime targets for combinatorial engineering. recently updated module boundary has been successful engineering short synthases, yet larger synthases constructed using have not investigated. Here we describe our design implementation a BioBricks-like platform to rapidly construct 5 triketide, 25 tetraketide, 125 pentaketide test every combination pikromycin synthase. Anticipated are detected from 60%...
The dehydratase (DH) domain of module 4 the 6-deoxyerythronolide B synthase (DEBS) has been shown to catalyze an exclusive syn elimination/syn addition water. Incubation recombinant DH4 with chemoenzymatically prepared anti-(2R,3R)-2-methyl-3-hydroxypentanoyl-ACP (2a-ACP) gave dehydration product 3-ACP. Similarly, incubation synthetic 3-ACP resulted in reverse enzyme-catalyzed hydration reaction, giving ∼3:1 equilbrium mixture 2a-ACP and a propionyl-SNAC (4), methylmalonyl-CoA, NADPH DEBS...
RifDH10, the dehydratase domain from terminal module of rifamycin polyketide synthase, catalyzes stereospecific syn dehydration model substrate (2S,3S)-2-methyl-3-hydroxypentanoyl-RifACP10, resulting in exclusive formation (E)-2-methyl-2-pentenoyl-RifACP10. RifDH10 does not dehydrate any other three diastereomeric, RifACP10-bound, diketide thioester substrates. On hand, when EryACP6, sixth erythromycin is substituted for RifACP10, stereospecifically dehydrates only...
The domains of modular polyketide synthases (PKSs) collaborate to extend and process intermediates; however, most their interactions with one another remain mysterious. We used AlphaFold 2 investigate how acyl carrier proteins (ACPs) present intermediates ketoreductases (KRs), processing capable not only setting the stereochemical orientations β-hydroxyl substituents but also α-substituents. In modules that do contain a dehydratase (DH), A- B-type KRs that, respectively, generate l-...
Large macromolecular assemblies are integral to most cellular processes, making their identification and structural characterization an important strategy for advancing our understanding of protein functions. In this pilot study, we investigated large multiprotein from the cytoplasm slime mold Dictyostelium discoideum using shotgun-electron microscopy (shotgun-EM), combined application mass spectrometry-based proteomics cryo-electron (cryo-EM) heterogenous mixtures proteins. With its...