full issue - Association of Biotechnology and Pharmacy

Current Trends in Biotechnology and Pharmacy
Vol. 5 (3) 1282-1297 July 2011, ISSN 0973-8916 (Print), 2230-7303 (Online)
1293
H-8). The presence of two peptide rings in the
actinomycin structure was established by the
presence of 3 amide signals at δ 8.19, 8.01 and
7.73. Also, the methyl groups of valine and
sarcosine amino acids gave the signals at δ 2.9
(one methyl) and 2.86 (two methyl groups).
Fig. 7. Growth and anticandidal activity of N.luteus as
affected by the agitation rate.
Isolation, purification and identification of the
anticandidal component (s) : The TLC
chromatogram for ethyl acetate extract of the
culture filtrate of Nocardioides luteus developed
as 6 major bands. By testing their activity against
C.albicans using paper disc assay method, they
all showed considerable activity as shown in
Table 3.
The compound was subjected several times
to chromatographic purification but it could not
be obtained as a pure sample. The semi-pure
compound was identified from its UV, IR, 1 H-
NMR and 13 C-NMR spectra and comparing these
spectra with published data (41) as derivative of
the actinomycins class of compounds. Its UV
spectrum showed maximum absorption (ë max
in
CH 3
OH) at 224, 427 and 444 nm.
Its IR spectrum showed the characteristic
peaks of NH group at 3433 cm -1 , aliphatic
hydrogens (CH, CH 2
and CH 3
) at 2966, 2924 and
2855 cm -1 . The amide group (CONH) signal
appeared at 1648 cm -1 .
The 1 H-NMR measurement showed the
characteristic signals of the phenoxazone
chromophore at δ 2.21 (CH 3
at position 4), 2.52
(CH 3
at position 6), 7.34 (d, H-7) and 7.61 (d,
The 13 C-NMR measurement showed the
characteristic signals of the phenoxazone
chromophore at δ 7.8 (CH 3
at position 4) and δ
15.11 (CH 3
at position 6), in addition to the
carbonyl group at δ 173.39 (C-3). The carbonyl
signals in between δ110-140 were attributed to
the other carbons of the phenoxazone group. The
signals corresponding to the amide and lactone
groups of the peptide system were detected in
between δ 160-172
The actinomycins are a family of
chromopeptide antibiotics, differing only in the
amino acids present in the pentapeptide chains,
which are elaborated by certain members of order
Actinomycetales. Actinomycin was first isolated
from a culture of Streptomyces antibioticus by
Waksman and Woodruff (40,41). It was the first
antibiotic to be produced after penicillin. The
various products have been designated A, B, C,
D, I, J, and X(42,43,44). It has been shown,
however, that these actinomycins were not
homogeneous substances but consisted of a
number of closely related biologically active
components. Evidence also suggested that the
A, B, D, and X complexes might consist of
identical components but differ in the relative
amount of each component present. Structural
studies have revealed that the actinomycin
molecule is composed of two polypeptides linked
to a chromophoric quinonoid moiety which is
the same for all components. Differences in the
components of an actinomycin complex or of
different complexes are due to variations in the
number, arrangement, and kinds of amino acids
present in the peptides.
El-Refai et al