| Summary: | The purpose of this research was to characteriz e of Actinobacteria isolate
and evaluate their diversity and potential in vitro with ability in controlling fungal
test growth. Analysis of sequences PKS and NRPS gene was conducted and
identify of the antifungal subtances producing of selected isolate. The diversity
analysis of cajuput isolates were using the fingerprinting was performed by rep-
PCR with the BOXA1R primer. Targeting genes encoding PKS and NRPS were
used to screen the antifungal biosynthetic potential of the selected isolates.
Among 17 of a total 43 of isolates showing activity were screened for producing
antifungi substances. The similarity of DNA fingerprints by BOXA1R repetitive
sequences of the isolates were revealed two clusters based on index similarity
70% and 90%, respectively. Therefore, could be predicted that the isolates have
high diversity relationships and show a positive correlation morphologically type
with other isolates and ARDRA analysis. Analysis of the conserved KS domain
sequences of the of PKS biosynthetic genes from the selected isolates was
displayed highest sequence identity (59.03%) to geldanamycin from S.
geldanamycininus AB430980. In the NRPS A-domain phylogeny, fragments of
targeted gene appeared to be most closely related to the sequenced fragments of
S.olivoverticillatus (AB432652). Phylogenetic analysis of selected isolate
(GMR22) was revealed showed 96.38% similarity of S.vayuensis.The effect of
various carbon and nitrogen sources of production antifungal was showed that
starch as good carbon sources and KNO3 has been shown to be an excellent
source of nitrogen for the growth and antifungal production of isolate,
respectively. Analysis of the antifungal compound was conducted by reagent
showed that the compound belong to the macrolide amide group. Meanwhile,
analysis by LC-MS gave molecular ions at m/z 741.32 was predicted as a
molecular weight of these antifungal compound. The subtances was including
ansamycin group and the molecular structure was suggested similar with
naphthomycin K. The MIC value of the compound were 62.5 μg/mL against C.
albicans.
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