Endogenous Production and Vibrational Analysis of Heavy-Isotope-Labeled Peptides from Cyanobacteria

Ricca, John G.; Mayali, Xavier; Qu, Jing; Weber, Peter K.; Poirier, Gerald; Dufresne, Craig P.; Louda, J. William; Terentis, Andrew C.

doi:10.1002/cbic.202400019

Title: Endogenous Production and Vibrational Analysis of Heavy-Isotope-Labeled Peptides from Cyanobacteria

Journal Article · 04 February 2024 · ChemBioChem: a European journal of chemical biology

DOI: https://doi.org/10.1002/cbic.202400019 · OSTI ID:2504057

^[1]; Mayali, Xavier ^[2]; Qu, Jing ^[3]; Weber, Peter K. ^[2]; Poirier, Gerald ^[3]; Dufresne, Craig P. ^[4]; Louda, J. William ^[1];

^[1]

Florida Atlantic Univ., Boca Raton, FL (United States)
Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)
Univ. of Delaware, Newark, DE (United States)
Thermo Fisher Scientific, West Palm Beach, FL (United States)

Stable isotope labeling is an extremely useful tool for characterizing the structure, tracing the metabolism, and imaging the distribution of natural products in living organisms using mass-sensitive measurement techniques. In this study, a cyanobacterium was cultured in ¹⁵N/¹³C-enriched media to endogenously produce labeled, bioactive oligopeptides. The extent of heavy isotope incorporation in these peptides was determined with LC–MS, while the overall extent of heavy isotope incorporation in whole cells was studied with nanoSIMS and AFM-IR. Up to 98 % heavy isotope incorporation was observed in labeled cells. Three of the most abundant peptides, microcystin-LR (MCLR), cyanopeptolin-A (CYPA), and aerucyclamide-A (ACAA), were isolated and further studied with Raman and FTIR spectroscopies and DFT calculations. This revealed several IR and Raman active vibrations associated with functional groups not common in ribosomal peptides, like diene, ester, thiazole, thiazoline, and oxazoline groups, which could be suitable for future vibrational imaging studies. More broadly, this study outlines a simple and relatively inexpensive method for producing heavy-labeled natural products. Finally, manipulating the bacterial culture conditions by the addition of specific types and amounts of heavy-labeled nutrients provides an efficient means of producing heavy-labeled natural products for mass-sensitive imaging studies.

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Research Organization:: Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)

Sponsoring Organization:: USDOE; USDOE National Nuclear Security Administration (NNSA)

Grant/Contract Number:: AC52-07NA27344

OSTI ID:: 2504057

Report Number(s):: LLNL--JRNL-847009; 1071549

Journal Information:: ChemBioChem: a European journal of chemical biology, Journal Name: ChemBioChem: a European journal of chemical biology Journal Issue: 6 Vol. 25; ISSN 1439-4227

Publisher:: ChemPubSoc EuropeCopyright Statement

Country of Publication:: United States

Language:: English

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60 APPLIED LIFE SCIENCES
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density functional theory
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Title: Endogenous Production and Vibrational Analysis of Heavy-Isotope-Labeled Peptides from Cyanobacteria

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