Supplementary Information
Penicillimide, an open-chain hemisuccinimide from Okinawan marine-derived
Penicillium copticola
Ying-Yue Bu, Hiroyuki Yamazaki, Kazuyo Ukai, and Michio Namikoshi
Contents
Materials and Methods
General Experimental Procedure
S2
Material
S2
Antimicrobial assay
S3
Table S1
S3
S1
Materials and Methods
General Experimental Procedure
1
H and
13
C NMR spectra were recorded on a JNM-AL-400 NMR spectrometer
(JEOL) at 400 MHz for 1H and 100 MHz for
C in CD3OD (δH 3.31, δC 49.15) and
13
acetone-d6 (δH 2.05, δC 29.92 and 206.68). Mass spectra were obtained by a JEOL
JMS-MS 700 mass spectrometer (EI or FAB mode with m-nitrobenzyl alcohol or
glycerol as the matrix). UV spectra were measured on a Hitachi U-3310 UV-Visible
spectrophotometer (Tokyo, Japan) and IR spectra on a PerkinElmer Spectrum One
Fourier transform infrared spectrometer (Waltham, MA, USA). Optical rotations were
determined with a JASCO P-2300 digital polarimeter (Tokyo, Japan). Preparative HPLC
was carried out using the Toso CCPU system equipped with a Tosoh UV-8010 detector.
Penicillimide (1): pale yellow solids. UV (MeOH) λmax nm (log ε) 202 (3.69), 280 (2.80).
IR (KBr) νmax 3419, 1739, 1734, 1635, 1501, 1424, 1364, 1287, 1059, 1048 cm–1.
HREIMS (m/z) 299.0555 ([M]+; calcd for C13H14NO5Cl, 299.0561). 1H and
13
C NMR
(CD3OD and acetone-d6), see Tables 1.
Material
Potato dextrose (PD) broth and malt extract were purchased from BD (Franklin Lakes,
NJ, USA). Ebios was purchased from Asahi Food & Healthcare Co. Ltd. (Tokyo, Japan).
6 mm filter disc for paper disc assay was purchased from Advantec (Tokyo, Japan). All
other chemicals including organic solvent were purchased from Wako Pure Chemical
S2
Industries Ltd. (Osaka, Japan).
Antimicrobial assay
Antimicrobial activity was determined using Mucor hiemalis IAM 6088 and
Aspergillus fumigatus IAM 13869 with the paper disc method. Each sample in MeOH
was adsorbed to a sterile filter disc, and, after the evaporation of MeOH, the disc was
placed on an agar plate and incubated for 2 days at 25 ˚C. Amphotericin B was used as
positive control.
Table S1
Antifungal activities (inhibition zone, mm) against Mucor hiemalis and
Aspergillus fumigatus.
M. hiemalis
A. fumigatus
compound
40 μg/disc
80 μg/disc
40 μg/disc
80 μg/disc
1
—a
—
—
—
2
7
9
7
10
3
—
9
—
9
4
11
15
9
16
5
—
—
—
—
6
8
10
11
13
Amphotericin B (10 μg/disc)
a
12
An inhibition zone was not detected.
S3
13