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Supplementary Files
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Chlorinated azaphilone pigments with antimicrobial and cytotoxic activities isolated from the deep sea derived Fungus Chaetomium sp. NA-S01-R1
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Weiyi Wang 1,*, Yanyan Liao 2, Ruixuan Chen 1, Yanping Hou 1, Wenqian Ke 1, Beibei Zhang 1, Maolin Gao 1, Zongze Shao 1, Jianming Chen 3,* and Fang Li 1,*
3
7
Content
8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40
Figure S1. 1H NMR spectrum of compound 1 Figure S2. 13C NMR spectrum of compound 1 Figure S3. HSQC spectrum of compound 1 Figure S4. HMBC spectrum of compound 1 Figure S5. HRESIMS spectrum of compound 1 Figure S6. 13C/DEPT spectrum of compound 1 Figure S7. 1H-1H COSY spectrum of compound 1 Figure S8. Selected NOESY spectrum of compound 1 Figure S9. Selected NOESY spectrum of compound 1 Figure S10. 1H NMR spectrum of compound 2 Figure S11. 13C NMR spectrum of compound 2 Figure S12. HSQC spectrum of compound 2 Figure S13. HMBC spectrum of compound 2 Figure S14. HRESIMS spectrum of compound 2 Figure S15. 13C/DEPT spectrum of compound 2 Figure S16. 1H-1H COSY spectrum of compound 2 Figure S17. NOESY spectrum of compound 2 Figure S18. 1H NMR spectrum of compound 3 Figure S19. 13C NMR spectrum of compound 3 Figure S20. HSQC spectrum of compound 3 Figure S21. HMBC spectrum of compound 3 Figure S22. HRESIMS spectrum of compound 3 Figure S23. 13C/DEPT spectrum of compound 3 Figure S24. 1H-1H COSY spectrum of compound 3 Figure S25. 1H NMR spectrum of compound 4 Figure S26. 13C NMR spectrum of compound 4 Figure S27. HSQC spectrum of compound 4 Figure S28. HMBC spectrum of compound 4 Figure S29. HRESIMS spectrum of compound 4 Figure S30. 13C/DEPT spectrum of compound 4 Figure S31. 1H-1H COSY spectrum of compound 4 Table S1; Table S2; Text S1
41 1.0
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
42 43 44
8.5
8.0
7.5
7.0
6.5
6.0
5.5
5.0
4.5
4.0
3.5
3.0
2.5
2.0
Chemical Shif t (ppm)
Figure S1. 1H NMR spectrum of compound 1
1.0 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0 -0.1
45 46 47
180
160
140
120
100
80
60
40
Chemical Shif t (ppm)
Figure S2. 13C NMR spectrum of compound 1
50
100
150
48 49 50
F2 Chemical Shif t (ppm)
6
5
4
3
Figure S3. HSQC spectrum of compound 1
2
1
50
100
150
51 52 53
54 55 56
F2 Chemical Shif t (ppm)
6
5
4
3
Figure S4. HMBC spectrum of compound 1
Figure S5. HRESIMS spectrum of compound 1
57 58 59
Figure S6. 13C/DEPT spectrum of compound 1
2
1
60 61 62
63 64 65
Figure S7. 1H-1H COSY spectrum of compound 1
Figure S8. Selected NOESY spectrum of compound 1
66 67 68
Figure S9. Selected NOESY spectrum of compound 1
0.20
0.15
0.10
0.05
69
0 13
12
11
70 71
10
9
8
7
6
5
4
3
2
Chemic al Shif t (ppm)
Figure S10. 1H NMR spectrum of compound 2 2.5
2.0
1.5
1.0
0.5
0
72 73 74
220
200
180
160
140
120
100
80
60
40
Figure S11. C NMR spectrum of compound 2 13
20
Chemic al Shif t (ppm)
0 20 40 60 80 100 120 140 160 180 200
75 76 77
F2 Chemical Shif t (ppm)
9
8
7
6
5
4
3
2
1
0
Figure S12. HSQC spectrum of compound 2
0
50
100
150
200
78 79 80
81 82 83
F2 Chemical Shif t (ppm)
9
8
7
6
5
4
3
Figure S13. HMBC spectrum of compound 2
Figure S14. HRESIMS spectrum of compound 2
2
1
0
84 85 86
Figure S15. 13C/DEPT spectrum of compound 2
87 88 89
90 91
Figure S16. 1H-1H COSY spectrum of compound 2
Figure S17. NOESY spectrum of compound 2
1.0
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
92
9
8
93 94
7
6
5
4
3
2
Chemical Shif t (ppm)
Figure S18. 1H NMR spectrum of compound 3
2.0
1.5
1.0
0.5
0
-0.5
95 96 97
200
180
160
140
120
100
80
60
40
Chemical Shif t (ppm)
Figure S19. 13C NMR spectrum of compound 3
50
100
150
200
98 99 100
F2 Chemical Shif t (ppm)
7
6
5
4
3
Figure S20. HSQC spectrum of compound 3
2
1
0
50
100
150
200
101 102 103
104 105 106
F2 Chemical Shif t (ppm)
7
6
5
4
3
2
Figure S21. HMBC spectrum of compound 3
Figure S22. HRESIMS spectrum of compound 3
107 108 109
Figure S23. 13C/DEPT spectrum of compound 3
1
0
110 111 112
Figure S24. 1H-1H COSY spectrum of compound 3
1.4 1.3 1.2 1.1 1.0 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1
113
0 10
9
8
114 115
7
6
5
4
3
2
Chemical Shif t (ppm)
Figure S25. 1H NMR spectrum of compound 4
2.0
1.5
1.0
0.5
0
116 117 118
220
200
180
160
140
120
100
80
60
40
Figure S26. 13C NMR spectrum of compound 4
20
Chemical Shif t (ppm)
0
50
100
150
200
119 120 121
F2 Chemical Shif t (ppm) 8
7
6
5
4
3
2
1
0
Figure S27. HSQC spectrum of compound 4
0
50
100
150
200
122 123 124
125 126 127
F2 Chemical Shif t (ppm)
7
6
5
4
3
2
Figure S28. HMBC spectrum of compound 4
Figure S29. HRESIMS spectrum of compound 4
1
0
128 129 130
Figure S30. 13C/DEPT spectrum of compound 4
131 132 133 134
Figure S31. 1H-1H COSY spectrum of compound 4 Table S1. Energies of the dominative conformers at MMFF94 force field of compound 1 Configuration
(1R, 7R, 8S, 8aR, 11S, 4’R, 5’S)-1
(1S, 7S, 8R, 8aS, 11S, 4’S, 5’R)-1
Conformer
Energy (kcal/mol)
1
131.80
2
133.62
3
133.64
4
133.71
1
134.38
2
135.20
3
135.64
4
135.94
5
135.95
6
136.04
7
135 136
137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155
136.16
Table S2. Energies of the conformers at B3LYP/6-311G** of compound 1 in methanol. Configuration
Conformer
Structure
E (Hartree) E (kcal/mol)Population (%)
(1R, 7R, 8S, 8aR, 11S, 4’R, 5’S)-1
3
-1879.82988243 -1179611.05
100.00
(1S, 7S, 8R, 8aS, 11S, 4’S, 5’R)-1
1
-1879.82899701 -1179610.50
26.34
(1S, 7S, 8R, 8aS, 11S, 4’S, 5’R)-1
2
-1879.82633158 -1179608.82
1.57
(1S, 7S, 8R, 8aS, 11S, 4’S, 5’R)-1
4
-1879.82994775 -1179611.09
72.10
Text S1: ITS1-5.8S-ITS2 rDNA sequence (MG786198) of strain Chaetomium sp. NA-S01-R1 LOCUS
Seq
568 bp
DNA
linear
PLN 11-JAN-2018
DEFINITION 1 Chaetomium sp. NA-S01-R1 18S ribosomal RNA gene, partial sequence; internal transcribed spacer 1, 5.8S ribosomal RNA gene, and internal transcribed spacer 2, complete sequence; and 28S ribosomal RNA gene, partial sequence. ACCESSION
Seq
VERSION KEYWORDS SOURCE
. Chaetomium sp.
ORGANISM Chaetomium sp. Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Sordariomycetes; Sordariomycetidae; Sordariales; Chaetomiaceae; Chaetomium. REFERENCE AUTHORS TITLE
1 (bases 1 to 568) Wang, W. Chlorinated azaphilone pigments with antimicrobial and cytotoxic
156 157 158 159 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199
activities isolated from the deep sea derived Fungus Chaetomium sp. NA-S01-R1 JOURNAL
Unpublished
REFERENCE
2 (bases 1 to 568)
AUTHORS TITLE
Wang, W. Direct Submission
JOURNAL
Submitted (11-JAN-2018) Key Laboratory of Marine Biogenetic
Resources, Third Institute of Oceanography, State Oceanic Administration, 178 Daxue Road, Xiamen, Fujian 361005, China COMMENT
Bankit Comment: ALT EMAIL:
[email protected].
Bankit Comment: TOTAL # OF SEQS:1.
##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES source
Location/Qualifiers 1..568 /organism="Chaetomium sp." /mol_type="genomic DNA" /isolate="NA-S01-R1" /isolation_source="seawater" /bio_material="Chaetomium sp." /db_xref="taxon:1769349" /collection_date="July 2017" /collected_by="Yanping Hou" /identified_by="Weiyi Wang"
misc_RNA
1..568 /note="contains 18S ribosomal RNA gene, partial sequence; internal transcribed spacer 1, 5.8S ribosomal RNA gene, and internal transcribed spacer 2, complete sequence; and 28S ribosomal RNA gene, partial sequence"
BASE COUNT
125 a
150 c
163 g
130 t
ORIGIN 1 ctggcggtct ctacttttcc gaggtcacct tgggttaaaa ggtggtttaa cggccggaac 61 ccgcggcgcg accagagcga gatgtatgct actacgctcg gtgcgacagc gagcccgcca 121 ctgcttttca gggcctgcgg cagccgcagg tccccaacac aagcccgggg gcttgatggt 181 tgaaatgacg ctcgaatagg catgcccgcc agaatgctgg cgggcgcaat gtgcgttcaa 241 agattcgatg attcactgaa ttaagcaatt cacattactt atcgcatttc gctgcgttct 301 tcatcgatgc cagaaccaag agatccgttg ttgaaagttt tgacttattc agtacagaag 361 actcagagag gccagaaatt atcaagagtt tggtgaccgc cggcgggcgc ccgcggtggg 421 gcccaggggc gcccggaggg taaaccccgg ggccgcccgc cgaagctacg gtataggtaa 481 cgttcacaat ggtttaggga gttttgcaac tctgtaatga tccctccgct ggttcaccaa 541 cggagacctt gttacgattt tttacttc //
