Supporting Information Metabolomics-driven Discovery of Meroterpenoids from a Musselderived Penicillium ubiquetum Thi Phuong Thuy oang,, Catherine Roullier,*,, Marie-Claude Boumard, Thibaut Robiou du Pont, assan Nazih, Jean-François Gallard, Yves François Pouchus, Mehdi A. Beniddir, livier Grovel, EA 2160 - Mer Molécules Santé, Université de Nantes, 44035 Nantes-cedex 1, France Phu Tho College of Pharmacy, 290000 Phu Tho, Vietnam Corsaire-ThalassMICS Metabolomics Facility, Biogenouest, Université denantes, France 1
Institut de Chimie des Substances Naturelles, CNRS UPR 2301, University of Paris-Saclay, 91198 Gif-sur-Yvette, France. Équipe Pharmacognosie-Chimie des Substances Naturelles BioCIS, Univ. Paris-Sud, CNRS, Université Paris-Saclay, 92290 Châtenay-Malabry, France 2
Table of contents Figure S1. Amounts of citrinin figured by their respective peak areas on the different culture media....3 Figure S2. PCA 2D score plot figuring all samples and including QC (grey stars)...4 Figure S3. 1 NMR spectrum (500 Mz, CDCl3) of 22-deoxyminiolutelide A (1)...5 Figure S4. 13 C NMR spectrum (125 Mz, CDCl 3 ) of 22-deoxyminiolutelide A (1)...6 Figure S5. CSY spectrum (500 Mz, CDCl 3 ) of 22-deoxyminiolutelide A (1)...7 Figure S6. SQC spectrum (500 Mz, CDCl 3 ) of 22-deoxyminiolutelide A (1)...8 Figure S7. MBC spectrum (500 Mz, CDCl 3 ) of 22-deoxyminiolutelide A (1)...9 Figure S8. NESY spectrum (500 Mz, CDCl 3 ) of 22-deoxyminiolutelide A (1)...10 Figure S9. Experimental ECD spectrum of 22-deoxyminiolutelide A (1) (Me)...11 Figure S10. 1 NMR spectrum (600 Mz, CDCl3) of 4-hydroxy-22-deoxyminiolutelide B (2)...12 Figure S11. 13 C NMR spectrum (150 Mz, CDCl3) of 4-hydroxy-22-deoxyminiolutelide B (2)...13 Figure S12. CSY spectrum (600 Mz, CDCl3) of 4-hydroxy-22-deoxyminiolutelide B (2)...14 Figure S13. SQC spectrum (600 Mz, CDCl3) of 4-hydroxy-22-deoxyminiolutelide B (2)...15 Figure S14. MBC spectrum (600 Mz, CDCl3) of 4-hydroxy-22-deoxyminiolutelide B (2)...16 Figure S15. RESY spectrum (600 Mz, CDCl3) of 4-hydroxy-22-deoxyminiolutelide B (2)...17 Figure S16. Experimental ECD spectrum of 4-hydroxy-22-deoxyminiolutelide B (2) (Me)...18 Figure S17. 1 NMR spectrum (500 Mz, CDCl3) of 22-deoxyminiolutelide B (3)...19 Figure S18. 13 C NMR spectrum (125 Mz, CDCl3) of 22-deoxyminiolutelide B (3)...20 Figure S19. Key MBC (left structure, single-headed arrows), CSY (bold lines) and NESY (right structure, double-headed arrows) correlations of 22-deoxyminiolutelide B (3)...21 Figure S20. CSY spectrum (500 Mz, CDCl3) of 22-deoxyminiolutelide B (3)...22 Figure S21. SQC spectrum (500 Mz, CDCl3) of 22-deoxyminiolutelide B (3)...23 Figure S22. MBC spectrum (500 Mz, CDCl3) of 22-deoxyminiolutelide B (3)...24 Figure S23. NESY spectrum (500 Mz, CDCl3) of 22-deoxyminiolutelide B (3)...25 Figure S24. Experimental ECD spectrum of 22-deoxyminiolutelide B (3) (Me)...26 Figure S25. 1 NMR spectrum (500 Mz, CDCl 3 ) of miniolutelide A (4)...27 Figure S26. 13 C NMR spectrum (125 Mz, CDCl 3 ) of miniolutelide A (4)...28 Figure S27. Experimental ECD spectrum of miniolutelide A (4) (Me)...29 Figure S28. 1 NMR spectrum (400 Mz, CDCl 3 ) of miniolutelide B (5)...30 3
Figure S29. 13 C NMR spectrum (100 Mz, CDCl 3 ) of miniolutelide B (5)...31 Figure S30. Experimental ECD spectrum of miniolutelide B (5) (Me)...32 Figure S31. 1 NMR spectrum (500 Mz, CDCl 3 ) of berkeleyacetal A (6)...33 Figure S32. 13 C NMR spectrum (125 Mz, CDCl 3 ) of berkeleyacetal A (6)...34 Figure S33. 1 NMR spectrum (600 Mz, CDCl 3 ) of miniolutelide C (7)...35 Figure S34. 13 C NMR spectrum (150 Mz, CDCl 3 ) of miniolutelide C (7)...36 Figure S35. 1 NMR spectrum (500 Mz, DMS) of 22-deoxy-10-oxominiolutelide B (8)...37 Figure S36. 13 C NMR spectrum (125 Mz, DMS) of 22-deoxy-10-oxominiolutelide B (8)...38 Figure S37. 1 NMR spectrum (600 Mz, CDCl 3 ) of eupeniacetal A (9)...39 Figure S38. 13 C NMR spectrum (150 Mz, CDCl 3 ) of eupeniacetal A (9)...40 Figure S39. Experimental ECD spectrum of eupeniacetal A (9) (Me)...41 Figure S40. Proposed biosynthetic pathway for isolated meroterpenoids based on Gu et al. (2018) and Iida et al. (2007), and observed production profiles clustered by Pearson correlation coefficient (r)...42 Figure S41. Comparison of PLC-RMS total ion chromatograms (TIC) together with extracted ion chromatograms for m/z 489.2119 of one replicate of MMS330 CYA-SW extract analyzed at different times (at 0, 10 and 40 days after extraction). Compounds 8 and 1 are highlighted...43 Figure S42. PLC-UV chromatogram (at 254 nm) of purified compound 8 after 2 h in C 3 CN/ 2 (40:60, v/v) shows conversion into compounds 1 and 3...44 Figure S43. (+)RESIMS/MS spectra of five ions from the molecular network...45 4
Peak area citrinin Media Figure S1. Amounts of citrinin figured by their respective peak areas on the different culture media. Peak areas were obtained by integration of the corresponding peak (at t R = 11.6 min) on extracted ion chromatograms for m/z = 251.0915 from PLC-(+)ESIMS data. Independent t-test analysis was performed in order to examine the significance of the differential production of citrinin between the two groups SW and DW on these three media (number of replicates = 3). A p-value of 0.0047 was obtained between CYA-DW and CYA-SW. 5
Figure S2. PCA 2D score plot figuring all samples and including QC (grey stars). The Quality Control (QC), which corresponded to a mix of all samples, was injected several times during the sequence of analyses. The plot shows that these profiles cluster together at the center enhancing the consistency of the results presented on the manuscript because very low deviation of the QC is observed. 6
1.06 1.07 1.04 1.03 1.09 3.01 1.05 1.10 1.11 1.40 0.82 3.01 5.66 3.09 3.23 3.16 1.08 3.04 6.28 6.05 6.04 5.88 5.75 4.27 4.25 4.24 4.23 3.78 3.27 3.26 3.25 3.23 3.09 3.07 2.91 2.90 2.89 2.24 2.21 1.62 1.52 1.44 1.43 1.31 1.23 1.21 1.17 1.14 0.75 2 1 16 17 18 26 3 4 5 15 14 25 19 20 12 13 24 6 7 8 11 10 22 23 9 21 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0 Chemical Shift (ppm) Figure S3. 1 NMR spectrum (500 Mz, CDCl3) of 22-deoxyminiolutelide A (1) 7
209.44 176.46 163.93 155.16 141.01 131.65 116.19 99.47 83.01 78.90 77.42 77.16 55.37 49.62 48.44 46.49 42.88 39.16 37.88 28.54 26.93 26.76 25.08 18.84 15.63 15.17 2 1 16 17 18 26 3 4 5 15 14 25 19 20 12 13 24 6 7 8 11 10 22 23 9 21 200 180 160 140 120 100 80 60 40 20 0 Chemical Shift (ppm) Figure S4. 13 C NMR spectrum (125 Mz, CDCl 3 ) of 22-deoxyminiolutelide A (1) 8
0 1 2 3 4 5 6 F1 Chemical Shift (ppm) 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0 F2 Chemical Shift (ppm) Figure S5. CSY spectrum (500 Mz, CDCl 3 ) of 22-deoxyminiolutelide A (1) 9
50 100 150 F1 Chemical Shift (ppm) 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0 F2 Chemical Shift (ppm) Figure S6. SQC spectrum (500 Mz, CDCl 3 ) of 22-deoxyminiolutelide A (1) 10
50 100 150 F1 Chemical Shift (ppm) 200 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0 F2 Chemical Shift (ppm) Figure S7. MBC spectrum (500 Mz, CDCl 3 ) of 22-deoxyminiolutelide A (1) 11
0 1 2 3 4 5 6 F1 Chemical Shift (ppm) 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0 F2 Chemical Shift (ppm) Figure S8. NESY spectrum (500 Mz, CDCl 3 ) of 22-deoxyminiolutelide A (1) 12
Figure S9. Experimental ECD spectrum of 22-deoxyminiolutelide A (1) (Me) 13
1.04 0.96 0.96 0.97 0.99 2.86 1.36 0.99 0.70 3.67 5.08 3.12 2.88 4.38 2.77 6.57 6.41 6.15 6.15 5.55 5.54 4.60 4.59 3.85 3.66 3.06 3.04 2.44 2.43 2.42 2.41 1.65 1.53 1.52 1.49 1.39 1.38 1.29 1.02 6 8 7 1 2 3 4 5 22 23 19 16 12 11 15 13 17 14 10 9 18 26 25 24 20 21 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 Chemical Shift (ppm) Figure S10. 1 NMR spectrum (600 Mz, CDCl3) of 4-hydroxy-22-deoxyminiolutelide B (2) 14
129.88 119.26 103.04 99.03 80.28 77.78 77.37 77.16 76.95 53.98 52.94 49.12 47.91 45.23 30.81 26.97 26.70 26.61 26.12 16.72 13.80 Me 6 8 7 1 2 3 4 5 22 23 19 16 12 11 15 13 17 14 10 9 18 25 24 20 21 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 Chemical Shift (ppm) Figure S11. 13 C NMR spectrum (150 Mz, CDCl3) of 4-hydroxy-22-deoxyminiolutelide B (2) 15
1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 F1 Chemical Shift (ppm) 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 F2 Chemical Shift (ppm) Figure S12. CSY spectrum (600 Mz, CDCl3) of 4-hydroxy-22-deoxyminiolutelide B (2) 16
0 20 40 60 80 100 F1 Chemical Shift (ppm) 120 140 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 F2 Chemical Shift (ppm) Figure S13. SQC spectrum (600 Mz, CDCl3) of 4-hydroxy-22-deoxyminiolutelide B (2) 17
20 40 60 80 100 120 140 160 180 200 F1 Chemical Shift (ppm) 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 F2 Chemical Shift (ppm) Figure S14. MBC spectrum (600 Mz, CDCl3) of 4-hydroxy-22-deoxyminiolutelide B (2) 18
1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 F1 Chemical Shift (ppm) 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 F2 Chemical Shift (ppm) Figure S15. RESY spectrum (600 Mz, CDCl3) of 4-hydroxy-22-deoxyminiolutelide B (2) 19
Figure S16. Experimental ECD spectrum of 4-hydroxy-22-deoxyminiolutelide B (2) (Me) 20
1.09 1.15 1.36 1.13 1.13 3.31 1.35 1.11 1.20 1.25 1.52 0.79 3.40 6.59 3.57 3.62 3.37 6.49 6.13 6.13 6.12 6.12 5.88 5.54 5.53 4.56 4.55 3.84 3.66 3.65 3.64 3.62 3.03 3.02 2.83 2.82 2.81 2.80 2.79 2.78 1.84 1.81 1.73 1.72 1.67 1.64 1.50 1.49 1.39 1.38 1.17 1.16 1.00 2 1 16 17 18 26 25 24 6 7 8 3 4 5 19 22 23 13 12 11 15 14 10 9 20 21 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0 Chemical Shift (ppm) Figure S17. 1 NMR spectrum (500 Mz, CDCl3) of 22-deoxyminiolutelide B (3) 21
180.33 175.41 163.46 157.60 134.89 129.53 120.68 102.85 99.09 82.35 80.84 77.81 77.42 77.16 76.91 61.11 52.83 49.27 46.72 45.20 43.83 43.52 34.82 27.32 26.76 26.21 18.79 17.51 13.79 2 1 16 17 18 26 25 24 6 7 8 3 4 5 19 22 23 13 12 11 15 14 10 9 20 21 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 Chemical Shift (ppm) Figure S18. 13 C NMR spectrum (125 Mz, CDCl3) of 22-deoxyminiolutelide B (3) 22
1 2 26 14 25 24 19 22 9 10 21 23 25 4 13 5 21 6β 19 6α 8 7 22 9 10 24 23 26 Figure S19. Key MBC (left structure, single-headed arrows), CSY (bold lines) and NESY (right structure, double-headed arrows) correlations of 22-deoxyminiolutelide B (3) 23
1 2 3 4 5 F1 Chemical Shift (ppm) 6 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 F2 Chemical Shift (ppm) Figure S20. CSY spectrum (500 Mz, CDCl3) of 22-deoxyminiolutelide B (3) 24
0 20 40 60 80 100 120 F1 Chemical Shift (ppm) 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 F2 Chemical Shift (ppm) Figure S21. SQC spectrum (500 Mz, CDCl3) of 22-deoxyminiolutelide B (3) 25
20 40 60 80 100 120 140 160 F1 Chemical Shift (ppm) 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 F2 Chemical Shift (ppm) Figure S22. MBC spectrum (500 Mz, CDCl3) of 22-deoxyminiolutelide B (3) 26
1 2 3 4 5 F1 Chemical Shift (ppm) 6 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 F2 Chemical Shift (ppm) Figure S23. NESY spectrum (500 Mz, CDCl3) of 22-deoxyminiolutelide B (3) 27
Figure S24. Experimental ECD spectrum of 22-deoxyminiolutelide B (3) (Me) 28
1.00 1.10 1.07 1.06 1.10 3.24 1.11 1.15 1.17 1.13 3.35 3.40 3.29 3.82 1.15 3.14 3.13 6.23 5.87 5.73 5.73 5.72 5.63 4.29 4.27 4.26 4.25 3.76 3.09 2.90 2.88 2.87 2.86 2.84 2.22 2.20 2.19 2.18 2.15 2.14 2.12 2.12 1.61 1.51 1.39 1.38 1.28 1.24 1.22 1.21 0.95 2 1 16 17 18 26 3 4 5 15 14 25 19 20 12 13 24 6 7 8 22 23 11 10 9 21 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 Chemical Shift (ppm) Figure S25. 1 NMR spectrum (500 Mz, CDCl 3 ) of miniolutelide A (4) 29
207.24 175.02 164.39 155.83 155.08 140.59 131.55 115.83 103.55 83.32 83.20 79.37 77.42 77.16 76.91 76.70 58.22 55.39 50.50 43.04 39.91 37.85 28.49 26.68 26.27 20.43 18.55 15.98 15.76 2 1 16 17 18 26 3 4 5 15 14 25 19 20 12 13 24 6 7 8 22 23 11 10 9 21 200 180 160 140 120 100 80 60 40 20 0 Chemical Shift (ppm) Figure S26. 13 C NMR spectrum (125 Mz, CDCl 3 ) of miniolutelide A (4) 30
Figure S27. Experimental ECD spectrum of miniolutelide A (4) (Me) 31
0.88 1.20 1.13 1.18 0.90 4.48 0.60 1.25 1.09 1.61 1.73 3.61 3.14 6.41 3.25 2.74 7.33 7.26 6.13 6.12 5.87 5.31 4.57 4.56 3.87 2.77 2.06 1.90 1.78 1.64 1.49 1.43 1.41 1.16 1.14 1.05 17 2 1 16 18 26 3 4 5 25 19 13 12 15 14 6 7 8 10 24 11 22 23 9 20 21 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0 Chemical Shift (ppm) Figure S28. 1 NMR spectrum (400 Mz, CDCl 3 ) of miniolutelide B (5) 32
179.23 174.01 163.52 157.46 134.92 129.78 120.80 108.44 103.51 82.47 80.50 79.62 78.08 77.48 77.16 76.85 67.84 52.96 52.52 50.64 43.46 41.95 38.32 27.34 26.21 19.38 18.68 18.02 13.64 17 2 1 16 18 26 3 4 5 25 19 13 12 15 14 6 7 8 10 24 11 22 23 9 20 21 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 Chemical Shift (ppm) Figure S29. 13 C NMR spectrum (100 Mz, CDCl 3 ) of miniolutelide B (5) 33
Figure S30. Experimental ECD spectrum of miniolutelide B (5) (Me) 34
1.28 1.35 0.86 3.25 0.81 2.03 0.88 0.85 0.99 3.12 3.36 1.85 0.69 3.13 3.54 3.28 2.85 6.14 6.13 6.12 6.11 6.05 6.04 4.54 4.53 4.52 4.50 3.87 3.86 3.85 3.41 3.38 2.80 2.80 2.77 2.76 2.50 2.49 2.47 2.10 2.09 2.07 2.06 1.79 1.67 1.64 1.54 1.51 1.43 1.42 1.37 0.78 17 1 2 16 18 26 3 4 5 25 19 12 15 14 13 24 6 7 8 11 22 23 9 10 20 21 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 Chemical Shift (ppm) Figure S31. 1 NMR spectrum (500 Mz, CDCl 3 ) of berkeleyacetal A (6) 35
204.46 175.99 171.13 167.85 136.54 135.71 129.17 127.41 99.55 83.10 77.41 77.16 76.90 76.11 62.48 56.73 53.49 52.36 43.35 38.58 35.82 35.02 30.37 29.19 27.67 26.26 21.32 19.36 16.23 17 1 2 16 18 26 3 4 5 25 19 12 15 14 13 24 6 7 8 11 22 23 9 10 20 21 200 180 160 140 120 100 80 60 40 20 0 Chemical Shift (ppm) Figure S32. 13 C NMR spectrum (125 Mz, CDCl 3 ) of berkeleyacetal A (6) 36
5.90 5.88 5.86 4.24 4.23 4.22 4.21 3.88 3.76 3.75 3.36 3.36 3.35 3.33 3.32 3.32 3.05 3.04 3.03 3.02 2.99 2.96 2.81 2.43 2.41 2.40 2.39 1.89 1.50 1.48 1.45 1.41 1.40 1.13 24 25 6 8 7 1 2 3 4 5 22 23 19 16 12 11 15 13 17 14 10 9 18 26 20 21 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 Chemical Shift (ppm) Figure S33. 1 NMR spectrum (600 Mz, CDCl 3 ) of miniolutelide C (7) 37
177.50 175.12 170.74 134.86 132.69 128.48 126.25 100.81 98.48 84.45 79.53 77.37 77.16 76.95 73.33 62.21 53.45 50.67 46.35 44.88 35.12 34.21 34.09 26.94 26.42 19.30 15.11 13.86 24 25 6 8 7 1 2 3 4 5 22 23 19 16 12 11 15 13 17 14 10 9 18 26 20 21 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 Chemical Shift (ppm) Figure S34. 13 C NMR spectrum (150 Mz, CDCl 3 ) of miniolutelide C (7) 38
6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0 Chemical Shift (ppm) Figure S35. 1 NMR spectrum (500 Mz, DMS) of 22-deoxy-10-oxominiolutelide B (8) 39
210.96 181.21 177.51 175.46 171.36 163.71 163.39 158.53 155.91 154.96 140.80 139.83 138.04 134.66 132.19 129.94 120.54 116.01 103.21 100.07 99.35 83.04 81.18 78.71 77.26 76.48 71.34 55.59 53.08 49.13 48.27 46.64 44.23 42.87 42.81 40.53 40.45 27.04 26.37 24.53 19.23 15.89 15.15 14.09 220 200 180 160 140 120 100 80 60 40 20 0 Chemical Shift (ppm) Figure S36. 13 C NMR spectrum (125 Mz, DMS) of 22-deoxy-10-oxominiolutelide B (8) 40
1.06 0.92 3.03 1.23 1.17 1.19 1.77 1.85 0.93 3.11 1.64 3.38 3.22 3.14 3.35 3.16 6.39 6.39 6.38 6.37 6.07 6.06 3.71 3.47 3.43 3.35 3.31 3.15 3.14 3.13 3.12 2.53 2.51 2.51 2.49 2.47 2.45 1.80 1.77 1.76 1.75 1.74 1.63 1.58 1.43 1.39 1.02 17 1 2 3 4 5 25 19 16 15 13 14 18 24 6 7 12 11 22 23 10 21 20 8 9 26 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0 Chemical Shift (ppm) Figure S37. 1 NMR spectrum (600 Mz, CDCl 3 ) of eupeniacetal A (9) 41
207.91 170.53 170.47 170.22 138.87 136.40 128.73 126.18 96.85 82.95 79.73 77.37 77.16 76.95 54.19 53.36 50.31 45.72 43.26 42.08 38.14 34.44 31.08 30.30 29.43 26.38 25.36 25.21 15.38 17 1 2 3 4 5 25 19 16 15 13 14 18 24 6 7 12 11 22 23 10 21 20 8 9 26 200 180 160 140 120 100 80 60 40 20 0 Chemical Shift (ppm) Figure S38. 13 C NMR spectrum (150 Mz, CDCl 3 ) of eupeniacetal A (9) 42
Figure S39. Experimental ECD spectrum of eupeniacetal A (9) (Me) 43
miniolutelide C (7) PP farnesyl pyrophosphate D E berkeleyacetal A (6) C 3,5-dimethyl orsellinic acid protoaustinoid A [ 2 ] + 2 [o] berkeleydione 22-epoxyberkeleydione [ 2 ] 22-deoxy-10-oxo miniolutelide B (8) D E D E eupeniacetal A (9) 22-deoxy miniolutelide A (1) 22-deoxy miniolutelide B (3) miniolutelide A (4) miniolutelide B (5) berkeleyacetal A (6) miniolutelide C (7) miniolutelide B (5) 22-deoxy miniolutelide B (3) 4-hydroxy-22-deoxy miniolutelide B (2) 22-deoxy-10-oxo miniolutelide B (8) miniolutelide A (4) eupeniacetal A (9) Legend CYA-DW CYA-SW DCA-DW DCA-SW MES-DW MES-SW Figure S40. Proposed biosynthetic pathway for isolated meroterpenoids based on Gu et al. (2018) and Iida et al. (2007), and observed production profiles clustered by Pearson correlation coefficient (r) 44
(x10,000,000) 1:TIC (1.00) 1:489.2119 (10.00) 5.0 4.0 8 T0 3.0 2.0 1.0 3 0.0 7.0 8.0 9.0 10.0 11.0 12.0 13.0 14.0 15.0 min (x10,000,000) 1:TIC (1.00) 1:489.2119 (10.00) 5.0 T0 + 10 days 4.0 3.0 8 2.0 1.0 3 1 0.0 7.0 8.0 9.0 10.0 11.0 12.0 13.0 14.0 15.0 min (x10,000,000) 1:TIC (1.00) 1:489.2119 (10.00) 5.0 4.0 3.0 8 T0 + 40 days 2.0 1.0 3 1 0.0 7.0 8.0 9.0 10.0 11.0 12.0 13.0 14.0 15.0 min Figure S41. Comparison of PLC-RMS total ion chromatograms (TIC) together with extracted ion chromatograms for m/z 489.2119 of one replicate of MMS330 CYA-SW extract analyzed at different times (at 0, 10 and 40 days after extraction). Compounds 8 and 1 are highlighted 45
mau 2500 8 2000 1500 1000 500 3 1 0 5 10 15 20 25 30 35 min Figure S42. PLC-UV chromatogram (at 254 nm) of purified compound 8 after 2 h in C 3 CN/ 2 (40:60, v/v) shows conversion into compounds 1 and 3 46
Inten. (x100,000) Berkeleyacetal A (6) C 26 32 8 t R 12.9 min 4.0 3.0 2.0 413.1908 427.2081 455.2077 395.1803 1.0 349.1634 441.1893 371.1507 198.9036223.1164 269.1043 0.0 150 200 250 300 350 400 450 m/z Miniolutelide C (7) C 26 32 9 t R 10.0 min Inten. (x100,000) 5.0 471.1959 4.0 3.0 429.1957 2.0 399.1809 1.0 457.1883 357.1817383.1789 411.1744 0.0 150 200 250 300 350 400 450 m/z Eupeniacetal A (9) C 26 32 8 t R 11.6 min Inten. (x100,000) 2.0 427.2144 455.2080 1.0 413.2025 224.1155 395.1736 0.0 150 200 250 300 350 400 450 m/z Inten. (x1,000,000) 7.5 5.0 487.1971 Miniolutelide B (5) C 26 32 10 t R 10.8 min 2.5 0.0 459.2033 441.1920 469.1885 506.2081 250 300 350 400 450 500 m/z 22-deoxyminiolutelide B (3) C 26 32 9 t R 10.5 min 1.5 1.0 0.5 Inten. (x1,000,000) 471.2045 453.2014 427.2204 0.0 150 200 250 300 350 400 450 500 m/z Figure S43. (+)RESIMS/MS spectra of five ions from the molecular network 47