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Population and Diversity of Endophytic Bacteria Associated with Medicinal Plant Curcuma zedoaria TRI RATNA SULISTIYANI; PUSPITA LISDIYANTI; YULIN LESTARI
Microbiology Indonesia Vol. 8 No. 2 (2014): June 2014
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (218.819 KB) | DOI: 10.5454/mi.8.2.4

Abstract

Traditionally Curcuma zedoaria (white turmeric) known as herbal medicine which possessing many biological activities. Many endophytic bacteria live in association with their host and may play an important biological roles. The main interest of this study was to investigate the endophytic bacterial diversity associated with white turmeric. White turmerics were collected from three locations in Bogor, West Java, Indonesia. The isolation of endophytic bacteria was carried out using 4 kinds media (Nutrient Agar (NA), NA contained white turmeric extract (NAT), Water Yeast Extract Agar (WYEA), WYEA contained white turmeric extract (WYEAT)), and 2 methods of spread plate and plant piece methods. The identification of selected isolates was conducted by molecular analysis based on 16S DNA. The suitable media and method of isolation endophytic bacteria were NA and spread plate method. A total of 207 bacterial colonies were isolated from rhizomes, stems, and leaves and 73 endophytic bacteria were selected based on morphological characteristics. From them, 32% isolates from Bojong Gede, 22% isolates from Cibinong and 46% isolates from Dramaga were obtained. Endophytic bacteria were predominated 38% in the rhizomes, 32% of stems, and 30% of leaves. Based on 16S rDNA sequence analysis, the isolates were belonging to the cluster Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria, with twenty three different genera includes Stenothropomonas, Pseudomonas, Enterobacter, Providencia, Klebsiella, Dickeya, Pantoea, Bacillus, Acinetobacter, Citrobacter, Mycobacterium, Cellulomonas, Microbacterium, Methylobacterium, Penylobacterium, Roseomonas, Agrobacterium, Bosea, Xanthobacter, Rhizobium, Burkholderia, Ralstonia, and Alcaligenes. The plant location, age, part of plant, media and method of isolation seem to influence the endophytic bacterial communities.
Cloning of a Transglutaminase Gene from Streptomyces thioluteus TTA 02 SDS 14 Seprianto Seprianto; Dewi Seswita Zilda; Yusro Nuri Fawzya; Suharsono Suharsono; Puspita Lisdiyanti; Agustinus Robert Uria
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 11, No 1 (2016): May 2016
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v11i1.189

Abstract

Microbial Transglutaminase (MTGase, EC 2.3.2.13) is an enzyme that catalyzes the transfer of acyl group. Many microbial strains that produce MTGase belong to Streptomyces members. This research was aimed at cloning of a MTGase gene. PCR–based screening of ten MTGase-producing streptomyces isolates from soil in West Nusa Tenggara led to detection of one potential isolate, designated as TTA 02 SDS 14. The partial  MTGase-encoding gene (470 bp)  was amplified by PCR and sequenced. The sequence result indicate its similarity of 93 % with that of Streptomyces cinnamoneus. The 16S rRNA gene analysis showed its identity as Streptomyces thioleteus. Fosmid-based construction of a genomic library from the isolate  and subsequent screening led to the isolation of  a ~40-Kb fosmid harboring a MTGase gene.
Screening of Marine Bacteria Capable of Degrading Various Polyaromatic Hydrocarbons Elvi Yetti; Ahmad Thontowi; Yopi Y; Puspita Lisdiyanti
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 10, No 3 (2015): December 2015
Publisher : Research and Development Center for Marine and Fisheries Product Processing and Biotechnol

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.v10i3.123

Abstract

This research aims to screen and to identify marine bacteria able to degrade Polyaromatic Hydrocarbons (PAHs). 7 bacterial isolates were screened for the ability to degrade 6 PAHs (phenanthrene, naphthalene, dibenzothiophene, fluorene, phenothiazine, and pyrene). This screening step was conducted by sublimation method, in which the color change or clear zone appearance was used as an qualitative indicator to show the capability of the selected isolate to degrade PAHs. Two bacterial isolates, designated as LBF-1-0102 and LBF1-0103, showed capability in degradating the PAHs tested. In particular, the isolate LBF-1-102 was potential in degrading phenanthrene, naphthalene, dibenzothiphene, fluorene, phenothiazine, and pyrene.The isolate LBF-1-0103  showed the ability in degrading 2 (two) PAHs, i.e. dibenzothiophene and  fluorene. Based on partial 16S rDNA analysis, LBF-1-0102 and LBF-1-0103 were identified as Pseudomonas balerica and Brachybacterium sp., respectively.
Antimicrobial Activity of Selenium Nanoparticles Synthesized by Actinomycetes Isolated from Lombok Island Soil Samples Shanti Ratnakomala; Nurul Fitri Sari; Fahrurrozi Fahrurrozi; Puspita Lisdiyanti
Jurnal Kimia Terapan Indonesia Vol 20, No 1 (2018)
Publisher : Research Center for Chemistry - LIPI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (546.255 KB) | DOI: 10.14203/jkti.v20i1.374

Abstract

AbstractA total of 98 actinomycetes were isolated from the soil and litter samples collected from the cacao and coffee plantation in Lombok Island, West Nusa Tenggara, Indonesia. These isolates were screened for their antimicrobial activity. Among 98 isolated strains, only 24 isolates showed antimicrobial activity against test microorganisms of which 20.4% were active against Bacillus subtilis BTCC B-612, 14.3% against Staphylococcus aureus BTCC B-611, and 5.1% against Escherichia coli BTCC B-609. Out of these 24 isolates, 3 were found to be able to grow in medium containing 3 mM Selenium oxide of which the culture were changed color to red. Two of the best strains, L-155 and L-156, were selected for assessing production of Selenium nanoparticles. Bioreduction of selenium nanoparticles was confirmed by UV–visible spectrophotometer which showed peak between 300 and 320 nm. Biosynthesized selenium nanoparticle from isolate actinomycetes L-155 and L-156 were found to have a broad spectrum of activity against the tested microorganisms: Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Micrococcus luteus, and Candida albicans. This study showed rapid and eco-friendly synthesis of selenium nanoparticles from soil actinomycetes. Most of these active isolates revealed to possess antibacterial property.
Purifikasi Inhibitor Atpase/Rna Helikase Virus Japanese Encephalitis Dari Streptomyces chartreusis Elfita, Lina; Ratnakomala, Shanti; Suryadi, Herman; Lisdiyanti, Puspita; Utama, Andi
Majalah Ilmu Kefarmasian Vol. 6, No. 2
Publisher : UI Scholars Hub

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Abstract

Japanese encephalitis virus (JEV) is a neuropathogenic virus commonly caused cen-tral nervous diseases such as meningitis and severe encephalitis. Although vaccine has been developed, no specific and effective drug is available so far. We previously carried out a screening of inhibitor of JEV RNA helicase, an enzyme that essential for virus replication, from Actinomycetes and found that Streptomyces chartreusis produce the inhibitor of JEV RNA helicase. In this study, an extracellular protein which has inhibition activity on ATPase activity of JEV RNA helicase was purified from supernatant of Streptomyces chartreusis culture by ammonium sulfate pre-cipitation and size exclusion chromatography. SDS-PAGE analysis showed a single band with aproximate molecular mass of 11,4 kDa, suggesting that the inhibitor was successfully purified into a single protein.
Enzymatic Hydrolysis of Mannan from Konjac (Amorphophallus sp.) Using Mannanase from Streptomyces lipmanii to Produce Manno-oligosaccharides Sasongko, Ashadi; Yopi,; Rahmani, Nanik; Lisdiyanti, Puspita; Saepudin, Endang
Makara Journal of Science Vol. 19, No. 3
Publisher : UI Scholars Hub

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Abstract

Mannan is an abundant polysaccharide that can be found in konjac (Amorphophallus sp.). Mannan can be enzymatically hydrolyzed using mannanase to produce manno-oligosaccharides which can be used as a prebiotic. The aims of this research are to determine the production time of mannanase from Streptomyces lipmanii, perform enzyme characterization, optimize the hydrolysis time, and characterize the hydrolysis product. A qualitative assay using the indicator Congo red showed that S. lipmanii generated a clear zone, indicating that S. lipmanii produced mannanase in konjac medium and possessed mannanolytic activity. Enzyme activity was determined through reducing sugar measurement using the dinitrosalycylic acid method, and optimum enzyme production was achieved at the second day of culture. Characterization of the enzyme showed that hydrolysis was optimum at pH 7 and at a temperature of 50 oC. The reducing sugar content was increased by an increasing the hydrolysis time, and reached an optimum time at 2 h. The degree of polymerization value of three was achieved after 2 h hydrolysis of mannan from konjac, indicating the formation of oligosaccharides. Analysis by thin layer chromatography using butanol, acetic acid, and water in a ratio of 2:1:1 as eluent showed the presence of compounds with a retention time between those of mannose and mannotetrose. Confirmation was also performed by HPLC, based on the retention time.
Beta-Glucosidase 1 (bgl1) Gene Analysis in Mutant and Wild-type of Penicillium sp. ID10-T065 Syafriana, Vilya; Nuswantara, Sukma; Mangunwardoyo, Wibowo; Lisdiyanti, Puspita
Jurnal Riset Biologi dan Aplikasinya Vol. 4 No. 1 (2022)
Publisher : Universitas Negeri Surabaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.26740/jrba.v4n1.p1-8

Abstract

In the previous study, Penicillium sp. ID10-T065 has been mutated using ultraviolet (UV), ethyl methyl sulfonate (EMS) and the combination of UV-EMS to increase Beta-glucosidase (bgl) activity. There were three mutants selected, UV13 (UV mutant), EM31 (EMS mutant), and UM23 (UV-EMS mutant). This study examined the mutations in the bgl gene encoding (bgl1) as well as sequence differences between mutants and wild-type of Penicillium sp. ID10-T065. The gene analysis was performed by PCR amplification and sequencing of the bgl1 gene. The results of bgl1 gene sequences (600 bp) from mutants were aligned with the wild-type, it was discovered that there were base alterations from position 2025 to 2050. Mutant UV13 showed the highest base alterations (7 bases) which occurred at position 2027 (T?C); 2036 (T?G); 2040 (T?G); 2047 (G?C); and 2048-2050 (TTG?GGA). Mutant EM31 showed alterations in five bases at positions 2034 (G?A), 2036 (T?G), 2037 (G?C), 2044 (G?C), and 2047 (G?T). Mutant UM23 showed two base alterations at position 2025 (T?A) and 2037 (G?C). UV irradiation and EMS mutation resulted in transition and transversion DNA, whereas the combination of UV-EMS mutation resulted in transversion mutations. Base alterations in UV13 and EM31 mutants, causing missense and silent mutation, while in UM23 mutant only silent mutations occur. The bgl1 gene analysis showed that mutation using UV light was more effective than using EMS or a combination of UV-EMS in Penicillium sp. ID10-T065.
The Marine Actinobacterium Streptomyces sp. BTA 1-131 as a Potential Producer of Anti-Nontuberculous Mycobacterial (Anti-NTM) Paramita, Ni Luh Putu Vidya; Atikana, Akhirta; Sukmarini, Linda; Untari, Febriana; Prayoga, Muhammad Bagas; Maulidia, Faiqoh Nur; Ratnakomala, Shanti; Lisdiyanti, Puspita; Nuryastuti, Titik; Hertiani, Triana
Squalen, Buletin Pascapanen dan Bioteknologi Kelautan dan Perikanan Vol 19, No 3 (2024): December 2024
Publisher : :Agency for Marine and Fisheries Research and Human Resources, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/squalen.882

Abstract

Nontuberculous mycobacteria (NTM) are environmental microorganisms, also known as opportunistic pathogens, found in patients with pulmonary tuberculosis. The emergence of antibiotic resistance is increased by prolonged antibiotic treatment for NTM infections. Therefore, alternative sources of new antibiotics are essential for the treatment of NTM infections. A marine actinobacterium, Streptomyces sp. BTA 1-131, isolated from a marine sponge, Melophlus sarasinorum, has been reported as a potential source of antibacterials and anticancer agents. The present study aimed to investigate the potential of Streptomyces sp. BTA 1-131 against two NTMs: Mycobacterium smegmatis and Mycobacterium fortuitum. Streptomyces sp. BTA 1-131 was fermented in three cultivation media (SYP, ISP2, and YS), and the secondary metabolites were extracted using methanol. The bioactivity screening showed inhibition of all methanolic extracts against the growth of M. smegmatis and M. fortuitum. The methanolic extract, which could inhibit both mycobacteria, was a crude extract derived from SYP liquid medium fermentation. The isolated compounds in this study were preliminarily identified using thin-layer chromatography (TLC). The TLC results showed different potential compounds in the crude extracts of Streptomyces sp. BTA 1-131 also highlighted the impact of the fermentation medium on the production of metabolites from Streptomyces. This study also added knowledge about the importance of the Indonesian marine actinobacterium Streptomyces sp. BTA 1-131 as a promising producer of anti-NTM compounds.
UTILIZATION OF INDONESIAN STRAIN ACTINOBACTERIA TO FACE THE THREAT OF BIOFILM AS THE MAIN CAUSE OF HEALTH CARE ASSOCIATED INFECTION Cahyo, Bian Dwi; Retnowati, Dwi; Lisdiyanti, Puspita
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 12 No. 2 (2025)
Publisher : BRIN - Badan Riset dan Inovasi Nasional

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.55981/jbbi.2025.13111

Abstract

Health Care Associated Infection (HCAI) is one of the global infectious diseases that is expected to cause around 10 million deaths by 2050. One of the main causes is biofilm, an exopolysaccharide layer formed by bacteria, often found on medical equipment such as catheters, and has high resistance to antibiotics. Prevention efforts can be made through the search for antibiofilm compounds. Actinobacteria are known to produce potential bioactive compounds. This study utilized Actinobacteria strains from the waters of Bitung, North Sulawesi (code BT-023-026) to explore their antibiofilm potential through isolation, laboratory tests, and genetic analysis, followed by expert confirmation in the fields of intelligence, BPOM, and microbiology. The results show the great potential of BT-023-026 as an antibiofilm agent to prevent antibiotic resistance due to HCAI, while playing a strategic role in biodefense and strengthening health security through early warning and early detection.
UTILIZATION OF INDONESIAN STRAIN ACTINOBACTERIA TO FACE THE THREAT OF BIOFILM AS THE MAIN CAUSE OF HEALTH CARE ASSOCIATED INFECTION Cahyo, Bian Dwi; Retnowati, Dwi; Lisdiyanti, Puspita
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 12 No. 2 (2025)
Publisher : BRIN - Badan Riset dan Inovasi Nasional

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.55981/jbbi.2025.13111

Abstract

Health Care Associated Infection (HCAI) is one of the global infectious diseases that is expected to cause around 10 million deaths by 2050. One of the main causes is biofilm, an exopolysaccharide layer formed by bacteria, often found on medical equipment such as catheters, and has high resistance to antibiotics. Prevention efforts can be made through the search for antibiofilm compounds. Actinobacteria are known to produce potential bioactive compounds. This study utilized Actinobacteria strains from the waters of Bitung, North Sulawesi (code BT-023-026) to explore their antibiofilm potential through isolation, laboratory tests, and genetic analysis, followed by expert confirmation in the fields of intelligence, BPOM, and microbiology. The results show the great potential of BT-023-026 as an antibiofilm agent to prevent antibiotic resistance due to HCAI, while playing a strategic role in biodefense and strengthening health security through early warning and early detection.