<p> Supporting Information for</p><p>Terpenoids Produced by Actinomycetes: Isolation, Structural Elucidation and </p><p>Biosynthesis of New Diterpenes, Gifhornenolone A and B from Verrucosispora gifhornensis</p><p>Masato Shirai1, Masaaki Okuda1, Keiichiro Motohashi1, Masaya Imoto2, Kazuo Furihata3, Yoshihide Matsuo,4 Atsuko Katsuta5, Yoshikazu Shizuri5 and Haruo Seto1*</p><p>1Faculty of Applied Bioscience, Tokyo University of Agriculture, Setagaya-ku, Tokyo 156-8502, Japan 2Science and Technology, Keio University, Yokohama-shi, Kanagawa, 223-8522, Japan 3Graduate School of Agricultural and Life Sciences, University of Tokyo, Bunkyo-ku, Tokyo, 113-8657, Japan 4Marine Biotechnology Institute Co. Ltd., Heita, Kamaishi-shi, Iwate, 026-0001, Japan (Current address: Suntory Holdings Limited, R&D Planning Division, Wakayamadai, Shimamoto-cho, Mishima-gun, Osaka, 618-8503, Japan) 5Marine Biosciences, Kamaishi Research Laboratory, Kitasato University, Heita, Kamaishi, Iwate, 026-0001, Japan </p><p>┴</p><p>1 Table of Contents: gifhornenolone A (1) P2-12, gifhornenolone B (2) P13-20, cyperusol C (3) P21-31</p><p>Spectral Data for gifhornenolone A (1)</p><p>1 Figure S1. H-NMR spectrum (400 MHz) of gifhornenolone A (1) in CDCl3 3</p><p>13 Figure S2. C-NMR spectrum (100 MHz) of gifhornenolone A (1) in CDCl3 4</p><p>Figure S3. HSQC spectrum (500 MHz) of gifhornenolone A (1) in CDCl3 5</p><p>Figure S4. HMBC spectrum (500 MHz) of gifhornenolone A (1) in CDCl3 6</p><p>Figure S5. COSY spectrum (500 MHz) of gifhornenolone A (1) in CDCl3 7</p><p>Figure S6. NOESY spectrum (500 MHz) of gifhornenolone A (1) in CDCl3 8</p><p>Figure S7. HR-MS spectrum of gifhornenolone A (1) 9</p><p>Figure S8. IR spectrum of gifhornenolone A (1) 9</p><p>13 13 Figure S9. C-NMR spectrum (150 MHz) of gifhornenolone A (1) labeled from [1- C] glucose in CDCl3 10</p><p>13 13 Figure S10. C-NMR spectrum (150 MHz) of gifhornenolone A (1) labeled from [U- C6] glucose in CDCl3 11</p><p>2 1 Figure S1. H-NMR spectrum (400 MHz) of Compound A (1) in CDCl3</p><p>1 Figure S1. H-NMR spectrum (400 MHz) of gifhornenolone A (1) in CDCl3</p><p>17</p><p>12 CH3 15 18 11 13 16 H3C H 14 O 1 9 2 H 10 8 3 7 5 6 4</p><p>HO 19</p><p>3 4 13 Figure S2. C-NMR spectrum (100 MHz) of gifhornenolone A (1) in CDCl3</p><p>17</p><p>12 CH3 15 18 11 13 16 H3C H 14 O 1 9 2 H 10 8 3 7 5 6 4</p><p>HO 19</p><p>5 Figure S3. HSQC spectrum (500 MHz) of gifhornenolone A (1) in CDCl3</p><p>17</p><p>12 CH3 15 18 11 13 16 H3C H 14 O 1 9 2 H 10 8 3 7 5 6 4</p><p>HO 19</p><p> methylene</p><p>6 Figure S4. HMBC spectrum (500 MHz) of gifhornenolone A (1) in CDCl3</p><p>17</p><p>12 CH3 15 18 11 13 16 H3C H 14 O 1 9 2 H 10 8 3 7 5 6 4</p><p>HO 19</p><p>7 Figure S5. COSY spectrum (500 MHz) of gifhornenolone A (1) in CDCl3</p><p>17</p><p>12 CH3 15 18 11 13 16 H3C H 14 O 1 9 2 H 10 8 3 7 5 6 4</p><p>HO 19</p><p>8 Figure S6. NOESY spectrum (500 MHz) of gifhornenolone A (1) in CDCl3</p><p>17</p><p>12 CH3 15 18 11 13 16 H3C H 14 O 1 9 2 H 10 8 3 7 5 6 4</p><p>HO 19</p><p>9 Figure S7. HR-MS spectrum of gifhornenolone A (1) Figure S8. IR spectrum of gifhornenolone A (1)</p><p>17</p><p>12 CH3 15 18 11 13 16 H3C H 14 O 1 9 2 H 10 8 3 7 5 6 4</p><p>HO 19</p><p>10 13 13 Figure S9. C-NMR spectrum (150 MHz) of gifhornenolone A (1) labeled from [1- C]glucose in CDCl3</p><p>17</p><p>12 CH3 15 18 11 13 16 H3C H 14 O 1 9 2 H 10 8 3 7 5 6 4</p><p>HO 19</p><p>11 13 13 Figure S10. C-NMR spectrum (150 MHz) of gifhornenolone A (1) labeled from [U- C6]glucose in CDCl3</p><p>17</p><p>12 CH3 15 18 11 13 16 H3C H 14 O 1 9 2 H 10 8 3 7 5 6 4</p><p>HO 19</p><p>12 Spectral Data for gifhornenolone B (2)</p><p>1 Figure S11. H-NMR spectrum (500 MHz) of gifhornenolone B (2) in CDCl3 13</p><p>13 Figure S12. C-NMR spectrum (150 MHz) of gifhornenolone B (2) in CDCl3 14</p><p>Figure S13. HSQC spectrum (400 MHz) of gifhornenolone B (2) in CDCl3 15</p><p>Figure S14. HMBC spectrum (500 MHz) of gifhornenolone B (2) in CDCl3 16</p><p>Figure S15. COSY spectrum (500 MHz) of gifhornenolone B (2) in CDCl3 17</p><p>Figure S16. NOESY spectrum (500 MHz) of gifhornenolone B (2) in CDCl3 18</p><p>Figure S17. HR-MS spectrum of gifhornenolone B (2) 19</p><p>13 1 Figure S11. H-NMR spectrum (400 MHz) of gifhornenolone B (2) in CDCl3</p><p>17</p><p>12 CH3 15 18 11 13 16 H3C H 14 O 1 9 2 H 10 8 3 7 5 6 HO 4 20 19 H3C OH</p><p>14 13 Figure S12. C-NMR spectrum (100 MHz) of gifhornenolone B (2) in CDCl3</p><p>17</p><p>12 CH3 15 18 11 13 16 H3C H 14 O 1 9 2 H 10 8 3 7 5 6 HO 4 20 19 H3C OH</p><p>15 Figure S13. HSQC spectrum (400 MHz) of gifhornenolone B (2) in CDCl3</p><p>17</p><p>12 CH3 15 18 11 13 16 H3C H 14 O 1 9 2 H 10 8 3 7 5 6 HO 4 20 19 H3C OH</p><p>16 Figure S14. HMBC spectrum (500 MHz) of gifhornenolone B (2) in CDCl3</p><p>17</p><p>12 CH3 15 18 11 13 16 H3C H 14 O 1 9 2 H 10 8 3 7 5 6 HO 4 20 19 H3C OH</p><p>17 Figure S15. COSY spectrum (500 MHz) of gifhornenolone B (2) in CDCl3</p><p>17</p><p>12 CH3 15 18 11 13 16 H3C H 14 O 1 9 2 H 10 8 3 7 5 6 HO 4 20 19 H3C OH</p><p>18 Figure S16. NOESY spectrum (500 MHz) of gifhornenolone B (2) in CDCl3</p><p>17</p><p>12 CH3 15 18 11 13 16 H3C H 14 O 1 9 2 H 10 8 3 7 5 6 HO 4 20 19 H3C OH</p><p>19 Figure S17. HR-MS spectrum of gifhornenolone B (2)</p><p>17</p><p>12 CH3 15 18 11 13 16 H3C H 14 O 1 9 2 H 10 8 3 7 5 6 HO 4 20 19 H3C OH</p><p>20 Figure S9. HR-MS spectrum of OxaloterpinFigure AS10. (1) HR-MS spectrum of Oxaloterpin A (1)</p><p>Spectral Data for cyperusol C (3)</p><p>1 Figure S18. H-NMR spectrum (400 MHz) of cyperusol C (3) in CDCl3 21</p><p>13 Figure S19. C-NMR spectrum (100 MHz) of cyperusol C (3) in CDCl3 22</p><p>Figure S20. HSQC spectrum (500 MHz) of cyperusol C (3) in CDCl3 23</p><p>Figure S21. HMBC spectrum (500 MHz) of cyperusol C (3) in CDCl3 24</p><p>Figure S22. COSY spectrum (500 MHz) of cyperusol C (3) in CDCl3 25</p><p>Figure S23. NOESY spectrum (500 MHz) of cyperusol C (3) in CDCl3 26</p><p>Figure S24. HR-MS spectrum of cyperusol C (3) 27</p><p>13 13 Figure S25. C-NMR spectrum (100 MHz) of cyperusol C (3) labeled from [1- C] glucose in CDCl3 29</p><p>13 13 Figure S26. C-NMR spectrum (100 MHz) of cyperusol C (3) labeled from [U- C6] glucose in CDCl3 30</p><p>21 1 Figure S18. H-NMR spectrum (400 MHz) of cyperusol C (3) in CDCl3</p><p>HO 14 1 CH3 9 2 10 8 H 7 11 3 4 6 5 12 H3C OH 15 CH3 13</p><p>22 13 Figure S19. C-NMR spectrum (100 MHz) of cyperusol C (3) in CDCl3</p><p>HO 14 1 CH3 9 2 10 8 H 7 11 3 4 6 5 12 H3C OH 15 CH3 13</p><p>23 Figure S20. HSQC spectrum (500 MHz) of cyperusol C (3) in CDCl3</p><p>HO 14 1 CH3 9 2 10 8 H 7 11 3 4 6 5 12 H3C OH 15 CH3 13</p><p>24</p><p>Figure S21. HMBC spectrum (500 MHz) of cyperusol C (3) in CDCl3</p><p>HO 14 1 CH3 9 2 10 8 H 7 11 3 4 6 5 12 H3C OH 15 CH3 13</p><p>25 FigureFigure S1 4S1. COSY2. COSY spectrum spectrum (500 (500 MHz) MHz) of (3ofR cypersol)-3-chloro-6-hydroxy-8-methoxy- C (2) in CDCl3 α - lapachone (8) in CDCl3</p><p>Figure S22. COSY spectrum (500 MHz) of cyperusol C (3) in CDCl3</p><p>HO 14 1 CH3 9 2 10 8 H 7 11 3 4 6 5 12 H3C OH 15 CH3 13</p><p>26 Figure S23. NOESY spectrum (500 MHz) of cyperusol C (3) in CDCl3</p><p>HO 14 1 CH3 9 2 10 8 H 7 11 3 4 6 5 12 H3C OH 15 CH3 13</p><p>27 Figure S24. HR-MS spectrum of cyperusol C (3)</p><p>HO 14 1 CH3 9 2 10 8 H 7 11 3 4 6 5 12 H3C OH 15 CH3 13</p><p>28 Figure S25. 13C-NMR spectrum (100 MHz) of cyperusol C (3)</p><p>13 labeled from [1- C]glucose in CDCl3</p><p>HO 14 1 CH3 9 2 10 8 H 7 11 3 4 6 5 12 H3C OH 15 CH3 13</p><p>29 Figure S26. 13C-NMR spectrum (100 MHz) of cyperusol C (3)</p><p>13 labeled from [U- C6]glucose in CDCl3</p><p>HO 14 1 CH3 9 2 10 8 H 7 11 3 4 6 5 12 H3C OH 15 CH3 13</p><p>30</p>