This dataset comprises dissolved organic matter (DOM) composition from axenic and xenic cultures of Thalassiosira gravida that were cultivated at the Alfred-Wegener-Institute (Bremerhaven, Germany) in March, 2023. After a cell density of ~ 15.000 cells * mL-1 was reached, cultures were filtered through a 0.2 µm polycarbonate (PC) filter (Whatman) that was cleaned by soaking in 10 % hydrochloric acid (HCl, Merck suprapure) for at least 12 h and subsequently rinsing with ultrapure water (Merck Millipore MilliQ). Cultures were grown under two temperatures (9 °C, 13.5 °C) and two photoperiods (16:8 h, 24:0 h light:dark). 2 mL of the sample were filtered through a 0.2 µm regenerated cellulose (RC)-membrane syringe filter (Sartorius) after defrosting. Molecular composition data were acquired with Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) coupled to a reversed-phase liquid chromatography (RPLC) with negative electrospray ionization (ESI) according to Lechtenfeld et al., 2024 (doi: 10.1021/acs.est.3c07219). Measurements were performed on on a solariX XR, Bruker Daltonics, Billerica, U.S.A. at the Helmholtz Centre for Environmental Research (UFZ; Leipzig, Germany). Molecular formulas were assigned and filtered using UltraMassExplorer (Leefmann et., 2019; doi: 10.1002/rcm.8315). If the Total ion chromatogram (TIC) was much higher and/or different in certain retention time windows compared to other samples of the same treatment, the sample was excluded from the dataset. The aim of this study was to investigate responses of algal extracellular release and bacterial DOM transformation to marine heatwave-like conditions.