Nonribosomal peptide synthetases (NRPSs) are multifunctional enzymes consisting of catalytic domains. The substrate specificities of adenylation (A) domains determine the amino-acid building blocks to be incorporated during nonribosomal peptide biosynthesis. The A-domains mediate ATP-dependent activation of amino-acid substrates as aminoacyl-O-AMP with pyrophosphate (PPi) release. Traditionally, the enzymatic activity of the A-domains has been measured by radioactive ATP-[(32)P]-PPi exchange assays with the detection of (32)P-labeled ATP. Recently, we developed a colorimetric assay for the direct detection of PPi as a yellow 18-molybdopyrophosphate anion ([(P2O7)Mo18O54](4-)). [(P2O7)Mo18O54](4-) was further reduced by ascorbic acid to give a more readily distinguishable blue coloration. Here we demonstrate the lab protocols for the colorimetric assay of PPi released in A-domain reactions.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/978-1-4939-3375-4_5 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Engineering the future of medicine: Natural products, synthetic biology and artificial intelligence for next-generation therapeutics.
Clin Transl Med
February 2025
Synthetic Biology of Microbial Natural Products, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), PharmaScienceHub (PSH), Saarbrücken, Germany.
The eXchange Unit between Thiolation domains approach and artificial intelligence (AI)-driven tools like Synthetic Intelligence are transforming nonribosomal peptide synthetase and polyketide synthase engineering, enabling the creation of novel bioactive compounds that address critical challenges like antibiotic resistance and cancer. These innovations expand chemical space and optimize biosynthetic pathways, offering precise and scalable therapeutic solutions. Collaboration across synthetic biology, AI, and clinical research is essential to translating these breakthroughs into next-generation treatments and revolutionizing drug discovery and patient care.
View Article and Find Full Text PDFComplete genome sequence of the marine mangrove fungus Sarcopodium sp.QM3-1 confirmed its high potential for antimicrobial activity.
Mar Genomics
March 2025
Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, Ministry of Natural Resources, 178 Daxue Road, Xiamen 361005, China; Applied Technology Engineering Center of Fujian Provincial Higher Education for Marine Resource Protection and Ecological Governance, Xiamen Key Laboratory of Intelligent Fishery, School of Marine Biology, Xiamen Ocean Vocational College, Xiamen 361100, China; Co-Innovation Center of Jiangsu Marine Bioindustry Technology, Jiangsu Ocean University, Lianyungang 222005, China. Electronic address:
Mangroves, owing to their unique living environment, serve as an important source of natural bioactive compounds. Sarcopodium sp. QM3-1, a marine fungus isolated from mangrove sediments of Quanzhou Bay, exhibited antifungal activity against the plant pathogen Agrobacterium tumefaciens and Magnaporthe oryzae.
View Article and Find Full Text PDFNovel Insights into the Nobilamide Family from a Deep-Sea : Chemical Diversity, Biosynthesis and Antimicrobial Activity Towards Multidrug-Resistant Bacteria.
Mar Drugs
January 2025
Department of Ecosustainable Marine Biotechnology, Stazione Zoologica Anton Dohrn, Via A.F. Acton, 55, 80133 Naples, Italy.
With rising concerns about antimicrobial resistance, the identification of new lead compounds to target multidrug-resistant bacteria is essential. This study employed a fast miniaturized screening to simultaneously cultivate and evaluate about 300 marine strains for biosurfactant and antibacterial activities, leading to the selection of the deep-sea BCP32. The integration of tandem mass spectrometry molecular networking and bioassay-guided fractionation unveiled this strain as a prolific factory of surfactins and nobilamides.
View Article and Find Full Text PDFDeletion of bZIP Transcription Factor Reveals Specialized Metabolites Potentially Regulating Stress Response in .
J Fungi (Basel)
January 2025
State Key Laboratory of Microbial Diversity and Innovative Utilization, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
Fungal secondary metabolism (SM) is highly correlated with physiological processes that are typically regulated by pleiotropic regulators. In this study, we purposefully altered , a crucial regulator associated with oxidative stress in CGMCC 3.1066.
View Article and Find Full Text PDFSubtilosin A production is influenced by surfactin levels in .
Microlife
January 2025
DTU Bioengineering, Technical University of Denmark, 2800 Kgs Lyngby, Denmark.
Although not essential for their growth, the production of secondary metabolites increases the fitness of the producing microorganisms in their natural habitat by enhancing establishment, competition, and nutrient acquisition. The Gram-positive soil-dwelling bacterium, , produces a variety of secondary metabolites. Here, we investigated the regulatory relationship between the non-ribosomal peptide surfactin and the sactipeptide bacteriocin subtilosin A.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!