Supporting information
Lorneic acids C and D, new trialkyl-substituted aromatic acids isolated from a terrestrial Streptomyces sp.
Ritesh Raju,a,b Oleksandr Gromykoc, Viktor Fedorenkoc, Andriy Luzhetskyya,b and Rolf Müller*a,b
a
Department of Microbial Natural Products, Helmholtz-Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research
(HZI), Saarland University, Campus C2 3, 66123 Saarbrücken, Germany
b
Department of Pharmaceutical Biotechnology, Saarland University, Campus C2 3, 66123 Saarbrucken, Germany.
c
Department of Genetics and Biotechnology of Ivan Franko National University of L’viv, Grushevskogo st. 4, L’viv 79005, Ukraine.
Figure S1a 1H NMR (500 MHz, methanol-d4) spectrum of lorneic acid C (1) ..................................................... 2
Figure S1b. COSY (500 MHz, methanol-d4) spectrum of lorneic acid C (1)........................................................ 3
Figure S1c. HSQC (500 MHz, methanol-d4) spectrum of lorneic acid C (1) ........................................................ 4
Figure S1d. HMBC (500 MHz, methanol-d4) spectrum of lorneic acid C (1) ...................................................... 5
Figure S2a. 1H NMR (500 MHz, methanol-d4) spectrum of lorneic acid D (2) .................................................... 6
Figure S2b. COSY (500 MHz, methanol-d4) spectrum of lorneic acid D (2) ....................................................... 7
Figure S2c. HSQC (500 MHz, methanol-d4) spectrum of lorneic acid D (2) ........................................................ 8
Figure S2d. HMBC (500 MHz, methanol-d4) spectrum of lorneic acid D (2) ...................................................... 9
Figure S3. HRMS spectrum of lorneic acid C (1)................................................................................................ 10
Figure S4. HRMS spectrum of lorneic acid D (1) ............................................................................................... 11
Taxonomic Identification ................................................................................................................................... 11
Biological assay.................................................................................................................................................... 13
1
Figure S1a. 1H NMR (500 MHz, methanol-d4) spectrum of lorneic acid C (1)
2
Figure S1b. COSY (500 MHz, methanol-d4) spectrum of lorneic acid C (1)
3
Figure S1c. HSQC (500 MHz, methanol-d4) spectrum of lorneic acid C (1)
4
Figure S1d. HMBC (500 MHz, methanol-d4) spectrum of lorneic acid C (1)
5
Figure S2a. 1H NMR (500 MHz, methanol-d4) spectrum of lorneic acid D (2)
6
Figure S2b. COSY (500 MHz, methanol-d4) spectrum of lorneic acid D (2)
7
Figure S2c. HSQC (500 MHz, methanol-d4) spectrum of lorneic acid D (2)
8
Figure S2d. HMBC (500 MHz, methanol-d4) spectum of lorneic acid D (2)
9
Figure S3. HRMS spectrum of lorneic acid C (1)
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Figure S4. HRMS spectrum of lorneic acid D (2)
Taxonomic identification of strain 4-15
16S rRNA gene sequence
GGACGTGGCGCTCTGCTACCATGCAGATCGACGATGAAGCCCTTCGGGGTGGATTAGTGGCGAACGGGTGAGTACACGTGGGCAATCTGCCCTTCACTCTGGGA
CAAGCCCTGGAAACGGGGTCTAATACCGGATACCACTCCTGCCTGCATGGGCGGGGGTTGAAAGCTCCGGCGGTGAAGGATGAGCCCGCGGCCTATCAGCTTG
TTGGTGGGGTAATGGCCCACCAAGGCGACGACGGGTAGCCGGCCTGAGAGGGCGACCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCA
GCAGTGGGGAATATTGCACAATGGGCGAAAGCCTGATGCACCGACGCCGCGTGAGGGATGACGGCCTTCGGGTTGTAAACCTCTTTCAGCAGGGAAGAAGCGA
AAGTGACGGTACCTGCAGAAGAAGCGCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGCGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGAGCT
CGTAGGCGGCTTGTCACGTCGGATGTGAAAGCCCGAGGCTTAACCTCGGGTCTGCATTCGATACGGGCTAGCTAGAGTGTGGTAGGGGAGATCGGAATTCCTG
GTGTAGCGGTGAAATGCGCAGATATCAGGAGGAACACCGGTGGCGAAGGCGGATCTCTGGGCCATTACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACA
11
GGATTAGATACCCTGGTAGTCCACGCCGTAAACGTTGGGAACTAGGTGTTGGCGACATTCCACGTCATCGGTGCCGCAGCTAACGCATTAAGTTCCCCGCCTGGG
GAGCTCGGCCGCCTGGCTAAAACCTCAATGGATTGATTGGGGCCCCGTACAATCGGTCTGAGCATGTGTCTTATATCGACGCCACCCTGAGAATCTTATTAGAGG
TTTGATATATCCTTGAAAGCTATTAATATTTGTTGCTCTCCTCTGTTGGTTCGTATATACAGTAGGTGGATGGTCTAT
LOCUS
DEFINITION
ACCESSION
VERSION
KEYWORDS
SOURCE
ORGANISM
REFERENCE
AUTHORS
TITLE
JOURNAL
REFERENCE
AUTHORS
TITLE
JOURNAL
EU285473
1454 bp
DNA
linear
BCT 12-DEC-2007
Streptomyces virginiae strain XSD-128 16S ribosomal RNA gene,
partial sequence.
EU285473
EU285473.1 GI:162136070
.
Streptomyces virginiae
Streptomyces virginiae
Bacteria; Actinobacteria; Actinobacteridae; Actinomycetales;
Streptomycineae; Streptomycetaceae; Streptomyces.
1 (bases 1 to 1454)
Jiang,J., Cao,X. and Chen,F.
The phylogenetic analysis of the genus Streptomyces by comparing
16S ribosomal RNA
Unpublished
2 (bases 1 to 1454)
Jiang,J., Chen,F. and Cao,X.
Direct Submission
Submitted (17-NOV-2007) Xuzhou Normal University, The Key
Laboratory of Biotechnology for Medicinal Plants of Jiangsu
Province, Shanghai Road 101, Xuzhou, Jiangsu 221116, China
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Biological Assay
Cytotoxic activity: Human HCT-116 colon carcinoma cells (DSMZ, ACC 581) were seeded at 1.2 x 104 cells per well of 96-well plates (Corning,
CellBind) in 180 μl complete medium and directly treated with 1 and 2 at 1 and 10 µg/ml to assess acute cytotoxicity. After 2 d incubation, 20 μl
of 5 mg/ml MTT (thiazolyl blue tetrazolium bromide) in PBS was added per well and it was further incubated for 2 h at 37°C. The medium was
then discarded and cells were washed with 100 μl PBS before adding 100 μl 2-propanol/10 N HCl (250:1) in order to dissolve formazan granules.
The absorbance at 570 nm was measured using a microplate reader (EL808, Bio-Tek Instruments Inc.), and cell viability was expressed as
percentage relative to the respective control. As a result, all tested derivatives are not cytotoxic up to a concentration of 10 µg/ml.
Microbial susceptibility was assessed by determination of the minimal inhibitory concentration (MIC). Therefore, microorganisms
were incubated in EBS or Myc medium over night at 30 °C on a shaker. Microorganisms were seeded into 96-well plated, treated with
different dilutions of the compound and incubated over night at 30°C on a shaker. The viability of microbial cells was analyzed by
measurement of the absorbance at 600 nm on a plate reader.
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