[
ABSTRAKPenelitian bertujuan memperoleh identitas isolat-isolat actinomycetes dari Raja
Ampat, Papua penghasil antimikroba dan antikanker dengan aktivitas tertinggi.
Penapisan aktivitas antimikroba dilakukan menggunakan metode difusi agar
terhadap bakteri (Escherichia coli, Bacillus subtilis, Staphylococcus aureus,
Micrococcus luteus) dan khamir (Candida albicans dan Saccharomyces
cerevisiae). Metabolit dari isolat terpilih yang memiliki aktivitas antimikroba
tertinggi dianalisis lanjut. Nilai Minimum inhibitor concentration (MIC) dari
senyawa murni yang diproduksi isolat terpilih terhadap mikroba diukur
menggunakan metode mikrodilusi. Kebocoran asam nukleat dan protein dari
mikroba uji dideteksi dengan menggunakan metode spektrofotometri pada
panjang gelombang 260 dan 280 nm. Kebocoran urasil dianalisis menggunakan
HPLC. Perubahan morfologi diamati menggunakan scanning electron microscope
(SEM). Uji sitotoksik dilakukan terhadap 6 sel kanker (MCM-B2, Leukemia,
T47D, HeLa, A549, dan WiDr) dan satu sel normal (Vero) menggunakan metode
trypan blue dan MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide). Sebanyak sembilan isolat potensial penghasil antimikroba tertinggi
(BL-36-1, BL-20-2, BL-14-2, RC-SS-37-4, RC-SS-37-16, BL-22-1, BL-06-5, BL-
22-3, dan BL-22-5) diidentifikasi berdasarkan data sekuen penuh gen 16S rRNA
(± 1.500 PB). Hasil penelitian menunjukkan 44 % isolat actinomycetes memiliki
aktivitas antimikroba. Senyawa F.5.1 merupakan senyawa murni yang diproduksi
oleh isolat terpilih BL-22-5. Nilai MIC senyawa tersebut terhadap mikroba uji
berkisar 16 -- 64 µg/ml. Aktivitas antifungi senyawa F.5.1 lebih tinggi
dibandingkan antibakteri. Senyawa F.5.1 menyebabkan kebocoran protein, asam
nukleat, dan urasil pada mikroba uji. Nilai IC50 senyawa F.5.1 berkisar 0,02 --
3,81 µg/ml terhadap sel kanker dan ≥ 30.000 µg/ml untuk sel normal. Isolat RCSS-
37-4, RC-SS-37-16, BL-22-3, dan BL-22-5 teridentifikasi sebagai
Streptomyces costaricanus (100 %), Streptomyces costaricanus (99,8 %),
Streptomyces parvulus (98,6 %), dan Streptomyces badius (98,9 %). Lima isolat
teridentifikasi sebagai Streptomyces spp. (BL-36-1, BL-20-2, BL-14-2, BL-22-1
dan BL-06-5) dan menjadi kandidat spesies baru karena memiliki nilai homologi
yang rendah terhadap spesies terdekatnya (93,9% -- 97,4 %).
ABSTRACTThe objective of this research was to obtain the identity of actinomycetes isolates
from Raja Ampat, Papua-producing antimicrobial and anticancer with the highest
activity. Antimicrobial screening was conducted based on agar diffusion method
against bacteria (Escherichia coli, Bacillus subtilis, Staphylococcus aureus,
Micrococcus luteus) and yeast (Candida albicans and Saccharomyces cerevisiae).
Metabolite from selected isolates with the highest antimicrobial activity was
chosen for subsequent analysis. Minimum inhibitor concentration (MIC) value
against microbial tested of pure bioactive compound was determined using
microdilution method. Leakages of nucleic acids and proteins from microbial
tested were detected using UV-vis spectrophotometer method on 260 and 280 nm
wavelengths. Uracil Leakage was analyzed using HPLC. Morphological changes
were observed using a scanning electron microscope (SEM). Determination of
cytotoxic activity was conducted in 6 cell lines (MCM-B2, Leukemia, T47D,
HeLa, A549, and WiDr) and normal cell (Vero) using trypan blue and MTT (3-
(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) methods. Nine
selected isolates with the highest antimicrobial activity (BL-36-1, BL-20-2, BL-
14-2, RC-SS-37-4, RC-SS-37-16, BL-22-1, BL- 06-5, 22-3-BL and BL-22-5)
were identified based on full sequence of 16S rRNA gene (1,500 bp). The results
showed 44% of isolates of actinomycetes have antimicrobial activity. F.5.1 is a
compound produced by the selected isolate (BL-22-5). MIC values of this
compound against tested microbes in range of 16 -- 64 µg/ml. Antifungal activity
from F.5.1 compound was higher than antibacterial activity. F.5.1 compound
caused leakage of proteins, nucleic acids, and uracil on tested microbes. Inhibitor
concentration 50% (IC50) value in the range of 0.02 -- 3.81 µg/ml against cancer
cells and ≥ 30,000 µg/ml for normal cells. Isolate RC-SS-37-4, RC-SS-37-16, BL-
22-3, and BL-22-5 have been identified as Streptomyces costaricanus (100%),
Streptomyces costaricanus (99.8%), Streptomyces parvulus ( 98.6%), and
Streptomyces badius (98.9%), respectively. Five isolates (BL-36-1, BL-20-2, BL-
14-2, BL-22-1 and BL-06-5) were identified as Streptomyces spp. and can be
presumed as candidates for new species because of the low homology value to
their closest related species (93.9 % -- 97.4%).;The objective of this research was to obtain the identity of actinomycetes isolates
from Raja Ampat, Papua-producing antimicrobial and anticancer with the highest
activity. Antimicrobial screening was conducted based on agar diffusion method
against bacteria (Escherichia coli, Bacillus subtilis, Staphylococcus aureus,
Micrococcus luteus) and yeast (Candida albicans and Saccharomyces cerevisiae).
Metabolite from selected isolates with the highest antimicrobial activity was
chosen for subsequent analysis. Minimum inhibitor concentration (MIC) value
against microbial tested of pure bioactive compound was determined using
microdilution method. Leakages of nucleic acids and proteins from microbial
tested were detected using UV-vis spectrophotometer method on 260 and 280 nm
wavelengths. Uracil Leakage was analyzed using HPLC. Morphological changes
were observed using a scanning electron microscope (SEM). Determination of
cytotoxic activity was conducted in 6 cell lines (MCM-B2, Leukemia, T47D,
HeLa, A549, and WiDr) and normal cell (Vero) using trypan blue and MTT (3-
(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) methods. Nine
selected isolates with the highest antimicrobial activity (BL-36-1, BL-20-2, BL-
14-2, RC-SS-37-4, RC-SS-37-16, BL-22-1, BL- 06-5, 22-3-BL and BL-22-5)
were identified based on full sequence of 16S rRNA gene (1,500 bp). The results
showed 44% of isolates of actinomycetes have antimicrobial activity. F.5.1 is a
compound produced by the selected isolate (BL-22-5). MIC values of this
compound against tested microbes in range of 16 -- 64 µg/ml. Antifungal activity
from F.5.1 compound was higher than antibacterial activity. F.5.1 compound
caused leakage of proteins, nucleic acids, and uracil on tested microbes. Inhibitor
concentration 50% (IC50) value in the range of 0.02 -- 3.81 µg/ml against cancer
cells and ≥ 30,000 µg/ml for normal cells. Isolate RC-SS-37-4, RC-SS-37-16, BL-
22-3, and BL-22-5 have been identified as Streptomyces costaricanus (100%),
Streptomyces costaricanus (99.8%), Streptomyces parvulus ( 98.6%), and
Streptomyces badius (98.9%), respectively. Five isolates (BL-36-1, BL-20-2, BL-
14-2, BL-22-1 and BL-06-5) were identified as Streptomyces spp. and can be
presumed as candidates for new species because of the low homology value to
their closest related species (93.9 % -- 97.4%)., The objective of this research was to obtain the identity of actinomycetes isolates
from Raja Ampat, Papua-producing antimicrobial and anticancer with the highest
activity. Antimicrobial screening was conducted based on agar diffusion method
against bacteria (Escherichia coli, Bacillus subtilis, Staphylococcus aureus,
Micrococcus luteus) and yeast (Candida albicans and Saccharomyces cerevisiae).
Metabolite from selected isolates with the highest antimicrobial activity was
chosen for subsequent analysis. Minimum inhibitor concentration (MIC) value
against microbial tested of pure bioactive compound was determined using
microdilution method. Leakages of nucleic acids and proteins from microbial
tested were detected using UV-vis spectrophotometer method on 260 and 280 nm
wavelengths. Uracil Leakage was analyzed using HPLC. Morphological changes
were observed using a scanning electron microscope (SEM). Determination of
cytotoxic activity was conducted in 6 cell lines (MCM-B2, Leukemia, T47D,
HeLa, A549, and WiDr) and normal cell (Vero) using trypan blue and MTT (3-
(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) methods. Nine
selected isolates with the highest antimicrobial activity (BL-36-1, BL-20-2, BL-
14-2, RC-SS-37-4, RC-SS-37-16, BL-22-1, BL- 06-5, 22-3-BL and BL-22-5)
were identified based on full sequence of 16S rRNA gene (1,500 bp). The results
showed 44% of isolates of actinomycetes have antimicrobial activity. F.5.1 is a
compound produced by the selected isolate (BL-22-5). MIC values of this
compound against tested microbes in range of 16 -- 64 µg/ml. Antifungal activity
from F.5.1 compound was higher than antibacterial activity. F.5.1 compound
caused leakage of proteins, nucleic acids, and uracil on tested microbes. Inhibitor
concentration 50% (IC50) value in the range of 0.02 -- 3.81 µg/ml against cancer
cells and ≥ 30,000 µg/ml for normal cells. Isolate RC-SS-37-4, RC-SS-37-16, BL-
22-3, and BL-22-5 have been identified as Streptomyces costaricanus (100%),
Streptomyces costaricanus (99.8%), Streptomyces parvulus ( 98.6%), and
Streptomyces badius (98.9%), respectively. Five isolates (BL-36-1, BL-20-2, BL-
14-2, BL-22-1 and BL-06-5) were identified as Streptomyces spp. and can be
presumed as candidates for new species because of the low homology value to
their closest related species (93.9 % -- 97.4%).]
