Benzophenone 3 bp 3

Bacillus licheniformis strain BLC‐01 (KC660142) was cultured in marine broth media 2216 and incubated at 37°C. The broth culture, with an absorbance of 1.7 at OD 600 nm was passed through a 0.20 μm Millipore filter. The filtrate was centrifuged (3250g, 40 min) and freeze dried at −80°C to a fine powder. The powder was used to make concentrations of 2, 10, 20, and 40 mg/ml of aqueous extracts (AE) metabolites, respectively, in milli Q water. These AEs were added to cultures of the less UV resistant Marinobacterium stanieri strain MARIS‐02 (KC660134) and investigated for additive UV resistance effects on the bacterium.
For AE assays, B. licheniformis and M. stanieri isolates were cultured in marine broth media 2216 and incubated at 37°C. Absorbance values of cultures were recorded. Time of irradiance and concentration of extracts were the variables whereas 0 mg/ml of B. licheniformis metabolite extract was the negative control. The positive control comprised a 1 mg/ml solution of the commercial UV sunscreen Benzophenone‐3 (BP‐3) from Sigma Aldrich. The reconstituted culture was spread onto marine agar and irradiated. Experiments were performed in triplicate.

Triclosan (TCS), bisphenol A (BPA), and benzophenone-3 (BP-3) standards were all purchased from Sigma-Aldrich (St. Louis, MO, USA). HPLC-grade methanol and water, analytical-grade ammonium acetate, formic acid, andβ-Glucuronidase/arylsulfatase were provided by Merck (Darmstadt, Germany). ¹³C₁₂-triclosan was obtained from Cambridge Isotope Laboratories Inc. (Andover, MA, USA).

Benzophenone-3 (BP3; 98%, CAS No. 131-57-7) was obtained from Sigma-Aldrich (Taufkirchen, Germany). Bis(tri-n-butyltin) oxide (TBTO; 97%, CAS No. 56-35-9, abcr GmbH, Karlsruhe, Germany) was used as positive control. Ethanol (EtOH; ≥99.8%, Carl Roth GmbH & Co. KG, Karlsruhe, Germany) was used as solvent to spike BP3 in water samples for matrix-matched calibrations. Tetrachloroethylene (TCE; HPLC grade, ≥99.9%, Sigma-Aldrich, Taufkirchen, Germany) was used as extraction solvent and formic acid (FA; Biosolve BV, Valkenwaard, The Netherlands) for adjusting the pH of water samples. For the analytical system, acetonitrile (ACN; ULC/MS grade, ≥99.99%; Biosolve BV, Valkenwaard, The Netherlands), and MilliQ water (ultrapure water purification system arium 611DI, Sartorius AG, Göttingen, Germany), both containing 0.01% FA, were used. A synthetic salt mix (Pro-Reef salt, Tropic Marin, Prof. Dr. Biener GmbH, Wartenberg, Germany) was used for the preparation of artificial seawater. Further details on physicochemical properties [69 ,70 ,71 ] of the active ingredients (BP3, TBTO), and chemicals used in additional bioassays can be found in Tables S1 and S22.