Rehman et al. (2008) identified an endophyte (Neurospora crassa), and designated it as ZP5SE from N. foetida seeds, and was able to produce CPT, when
cultured in Sabouraud Dextrose Liquid Medium under the shake flask culturing
environments. This endophyte was capable to propagate in Sabouraud broth
(100 ml) poured in a 500 ml Erlenmeyer flask, and grown on a shaking incubator,
agitated at 220 rpm for a duration of 10 days at a temperature of 28 ± 2 °C. From
the growth kinetics study, it was evidenced that this fungal species, showing the
exponential phase of growth up to 7 days of culturing. The produced CPT content
by this endophyte was analyzed using High Performance Liquid Chromatograhy
(HPLC) followed by Liquid chromatography–mass spectrometry (LC/MS) and
Tandem mass spectrometry (MS/MS). The obtained CPT was tested against the
growth of human lung cancer (A-549) and ovarian cancer (OVCAR-5) cells. The
results were positive with effective inhibition of the growth of cancer cells, and the
observed data were comparable to the data obtained for the standard drug, CPT.
Another endophyte, Nodulisporium sp. was isolated from N. nimmoniana inner
barks by Rehman et al. (2009). This fungal species possessed thin hyphae, which
was found to extend up to 6.4 lm in diameter, and profuse conidiophores that were
branched, verticillately. The fungus, when grown on liquid culture (Sabouraud
broth) media produced CPT up to 5.5 lg/g DW of mycelia after one week. Further,
the authors have explored the possibility of its culturing in an airlift bioreactor with
5 to 18 l volume. They found that mycelium growth initiated within 5 h of inoculation, and the maximum growing was documented on Sixth day of culturing in a
bioreactor with the yield of 45 lg/g DW of CPT.
From the plant, Apodytes dimidiata, 2 strains of F. solani (MTCC 9667 and
MTCC 9668) were isolated, and these strains were shown to produce both CPT in
their mycelia. Interestingly, MTCC 9668 strain was also shown to produce
10-hydroxycamptothecin, however in low quantities in mycelia. These endophytes,
MTCC 9668 and MTCC 9667 after culturing on a liquid broth for 4 days could able
to produce 53 and 37 lg CPT/100 g DW of mycelia, respectively (Shweta et al.
2010). Likewise, the produced quantity of 9-methoxycamptothecin and
10-hydroxycamptothecin by MTCC 9668 was observed to be 44.9 and 8.2 lg/
100 g DW, correspondingly.
A study carried out by Musavi et al. (2015) identified the occurrence of an
endophyte, F. oxysporum NFX06 associated with N. nimmoniana. They evaluated
the production of cell-allied CPT from this endophytic fungus, and proposed its
growth kinetics. To determine the consequence of substrate levels, a model was
constructed by applying response surface methodology on the basis of central
composite design. They used dextrose, magnesium sulfate, and peptone as independent variables to evaluate the production of CPT under sub-merged fermentation conditions. The highest CPT yield noticed from the central composite design
was found to be 598 ng/g DW of mycelia. Also, the model-endorsed experimental
yield of CPT and the optimal expected yield of CPT from dried mycelia were
observed to be 610 ng/g and 628 ng/g DW at the substrate levels of 9.2 g/l peptone,
42.6 g/l dextrose, and 0.26 g/l MgSO 4 , correspondingly.
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M. K. Swamy et al.
cultured in Sabouraud Dextrose Liquid Medium under the shake flask culturing
environments. This endophyte was capable to propagate in Sabouraud broth
(100 ml) poured in a 500 ml Erlenmeyer flask, and grown on a shaking incubator,
agitated at 220 rpm for a duration of 10 days at a temperature of 28 ± 2 °C. From
the growth kinetics study, it was evidenced that this fungal species, showing the
exponential phase of growth up to 7 days of culturing. The produced CPT content
by this endophyte was analyzed using High Performance Liquid Chromatograhy
(HPLC) followed by Liquid chromatography–mass spectrometry (LC/MS) and
Tandem mass spectrometry (MS/MS). The obtained CPT was tested against the
growth of human lung cancer (A-549) and ovarian cancer (OVCAR-5) cells. The
results were positive with effective inhibition of the growth of cancer cells, and the
observed data were comparable to the data obtained for the standard drug, CPT.
Another endophyte, Nodulisporium sp. was isolated from N. nimmoniana inner
barks by Rehman et al. (2009). This fungal species possessed thin hyphae, which
was found to extend up to 6.4 lm in diameter, and profuse conidiophores that were
branched, verticillately. The fungus, when grown on liquid culture (Sabouraud
broth) media produced CPT up to 5.5 lg/g DW of mycelia after one week. Further,
the authors have explored the possibility of its culturing in an airlift bioreactor with
5 to 18 l volume. They found that mycelium growth initiated within 5 h of inoculation, and the maximum growing was documented on Sixth day of culturing in a
bioreactor with the yield of 45 lg/g DW of CPT.
From the plant, Apodytes dimidiata, 2 strains of F. solani (MTCC 9667 and
MTCC 9668) were isolated, and these strains were shown to produce both CPT in
their mycelia. Interestingly, MTCC 9668 strain was also shown to produce
10-hydroxycamptothecin, however in low quantities in mycelia. These endophytes,
MTCC 9668 and MTCC 9667 after culturing on a liquid broth for 4 days could able
to produce 53 and 37 lg CPT/100 g DW of mycelia, respectively (Shweta et al.
2010). Likewise, the produced quantity of 9-methoxycamptothecin and
10-hydroxycamptothecin by MTCC 9668 was observed to be 44.9 and 8.2 lg/
100 g DW, correspondingly.
A study carried out by Musavi et al. (2015) identified the occurrence of an
endophyte, F. oxysporum NFX06 associated with N. nimmoniana. They evaluated
the production of cell-allied CPT from this endophytic fungus, and proposed its
growth kinetics. To determine the consequence of substrate levels, a model was
constructed by applying response surface methodology on the basis of central
composite design. They used dextrose, magnesium sulfate, and peptone as independent variables to evaluate the production of CPT under sub-merged fermentation conditions. The highest CPT yield noticed from the central composite design
was found to be 598 ng/g DW of mycelia. Also, the model-endorsed experimental
yield of CPT and the optimal expected yield of CPT from dried mycelia were
observed to be 610 ng/g and 628 ng/g DW at the substrate levels of 9.2 g/l peptone,
42.6 g/l dextrose, and 0.26 g/l MgSO 4 , correspondingly.
318
M. K. Swamy et al.