Puspita Lisdiyanti’s research while affiliated with National Research and Innovation Agency and other places

Aims This study aims to assess the antimicrobial activity of sponges from Lembeh Strait, explore their prokaryotic diversity, and investigate correlations between sponge antimicrobial activities and their associated prokaryotic communities. Methods and Results Secondary metabolites were extracted from sixty two sponge tissues using MeOH:EtOAc as solvent (1:1). Antimicrobial activity of the sponge crude extracts was tested against Escherichia coli, Aeromonas salmonicida, Bacillus subtilis, Staphylococcus simulans, Saprolegnia parasitica, and Candida oleophila. The strongest antimicrobial activity was observed for sponge extracts from Theonella sp. against S. parasitica. Microbial community profiling was performed for sponges and environmental samples (seawater and sediment) using 16S rRNA gene amplicon sequencing. Distinct prokaryotic community compositions were observed in sponges compared to their environments, with a strong influence of host-identity. A significant positive correlation between the phylum Actinobacteriota and sponge antimicrobial activity suggests importance of Actinobacteriota for sponge's bioactive properties. In addition, six amplicon sequence variants (ASVs) belonging to the phyla Actinobacteriota, Chloroflexota, Rhodothermota, and candidate phylum PAUC34f showed significant positive correlations with particular sponge antimicrobial activities, indicates these taxa as interesting targets for future bioprospecting endeavours. Conclusions This study highlights sponges from the Lembeh Strait, Indonesia, as producers of antimicrobial compounds and the importance of host identity on sponge-prokaryotic community composition. Moreover, poorly studied taxa such as candidate phylum PAUC34f and Chloroflexota members might represent interesting targets for future bioprospecting endeavours.

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.

The World Health Organization (WHO) has determined a list of pathogens that require the development of new antimicrobials due to resistance problems; these include Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus. In addition, Mycobacterium smegmatis has been used for antimycobacterial discovery to address the increasing burden of tuberculosis. In this study, optimization of antimicrobial activity, secondary metabolite profiling, and strain identification was conducted on Actinobacteria InaCC A759. Intracellular and extracellular extracts of Actinobacteria InaCC A759 were found to have different antimicrobial activities. The minimum inhibitory concentration (MIC) values of the extract to inhibit the growth of M. smegmatis, E. coli, and P. aeruginosa were 50, 25, and 100 µg/mL (intracellular), and 25, 25, and 100 µg/mL (extracellular), respectively. However, neither extract was able to inhibit the growth of S. aureus. Metabolite profiling using High resolution‐mass spectrometry (HR‐MS) resulted in differences in the major compound between the two extracts of Actinobacteria InaCC A759, namely n‐acetyltyramine (C10H13NO2/179.0945) (24.24%) (intracellular) and palmitic acid (C16H32O2/273.27034) (86.92%) (extracellular). Based on molecular analysis of the 16S rRNA gene, Actinobacteria InaCC A759 is identical to the Streptomyces olivaceus strain FoRh46. The antimicrobial activity and secondary metabolites profiles of Streptomyces olivaceus InaCC A759 have not been previously reported.

Candida albicans and Staphylococcus aureus can coexist to form a biofilm, leading to infections associated with biofilms. Actinomycetes produce secondary metabolites known as antibiotics, antifungals, antibiofilm, anticancer, and antimalarials. This study was aimed to explore the antibiofilm activity of secondary metabolites of Actinomycetes InaCC A758 extracts (InaCC A758) against dual-species biofilms, i.e., C. albicans and S. aureus. Ethyl acetate and chloroform were used as solvents in a maceration extraction technique to isolate the compound designated InaCC A758. The fractionation of the InaCC A758 extracts was carried out using semi-preparative HPLC. Identification of the compounds from the InaCC A758 extracts was performed using gas chromatography-mass spectrometry analysis. Antimicrobial and antibiofilm testing of the InaCC A758 extracts was done utilizing the micro broth dilution method. The morphology of the biofilms following treatment with the InaCC A758 extracts was visualized using scanning electron microscopy (SEM). The minimum inhibitory concentration of InaCC A758 against dual-species biofilms was 400 μg/ml. The concentrations of InaCC A758 required to inhibit 50% and 80% of dual-species biofilm formation (BIC50 and BIC80) ranged from 3.57–6.66 and 29.25–30.50 μg/ml, respectively. The concentrations needed to reduce formed dual-species biofilms by 50% and 80% (BRC50 and BRC80) ranged from 100.62–131.85 and 596.4–849.6 μg/ml, respectively. SEM observation showed a reduced number of cells and disruption of the cell membranes when exposed to the InaCC A758 extracts. The InaCC A758 contains seven compounds, i.e., dodecanoic acid 3-hydroxy-; 1,3,5-pentanetriol,3-methyl-; 1,3-dioxolane-4-methanol, 2-ethyl-; 2-cyclopropylcarbonyloxytridecane; 2-hepten-1- ol,(E); methyl 6-methyl heptanoate; and 11-octadecenoic acid, methyl ester. The InaCC A758 extract altered cell morphology and exhibited potential as an anti-biofilm agent, particularly against dual-species biofilms.

The phylum Actinobacteria recognized as the most promising producers of many bioactive compounds. Among actinobacteria, the genus Streptomyces are widely known as prolific producers of many antibiotics, for example Streptomycin. The present study investigates potential bioactivity of actinobacteria BTA 1-131 isolated from an Indonesian marine sponge Melophlus sarassinorum. Molecular identification using the 16S rRNA gene sequencing showed that the isolate is similar to Streptomyces kunmingensis (98.4%). The isolate was cultivated on to three different fermentation media (ISP2, YS, and SYP) to select the best media for bioactive compounds production. The compounds were extracted from both solid and liquid cultures using methanol as a solvent. The crude methanol extracts were tested for antibacterial and anticancer activity. The bioactivity screening indicates active potential of the extracts against bacterial pathogen Staphylococcus aureus ATCC 13420, invasive breast cancer MDA-MB-231 and human colorectal adenocarcinoma cells CaCo-2. The extracts profiling using LC-MS/MS method indicates probable influence among different fermentation media to selectively induce production of target bioactive compounds. Although further compounds dereplication and fractionation are important for specific identification, the present study demonstrates the potential of Streptomyces BTA 1-131 as producer of tetracycline and/or staurosporine. In addition to that, further genome-mining approach and analysis using a whole genome sequencing data also important to comprehensively explore the bioactive potential of the actinobacteria BTA 1-131.

Currently, the study and the development of plastic degradation are receiving great attention since it could potentially be applied to plastic degradation. Owing to its environmentally benign nature, the enzymatic process of plastic degradation is highly desired. Polyethylene terephthalate (PET) is one of the most common plastic materials. The degradation of PET was possible by the treatment of the three hydrolases, namely cutinase, lipase, and esterase. On the other hand, marine actinomycetes, which are commonly found to associate with other organisms such as a sponge or residing in a unique environment such as marine sediment, could be a great source of valuable metabolites and enzymes. For that reason, in this study, six isolates of marine actinomycetes showing lipase activity were tested for PET-film degradation. The PET-film degradation was performed by incubating the isolates in a liquid media supplemented with PET films for 14 days. The resulting PET films were then subjected to XRD, FTIR, and SEM analysis. Despite of the decrease in crystallinity of PET-film which was only observed in treatment with isolate BLC 17-1, the FTIR analysis revealed that all absorbances, which are related to PET functionalities, were reduced. Further analysis by SEM showed that the surface of PET films treated with isolates was damaged, indicating degradation of the films. According to the results, it is concluded that the six isolates of marine actinomycetes have the potential to be applied to PET-film degradation.

Pahira DSJ, Damayanti E, Lisdiyanti P, Mustofa, Hertiani T. 2023. Exploring the potency of Streptomyces koyangensis strain SHP 9-3 isolated from the soil of Enggano Island (Indonesia) as an antibacterial source. Biodiversitas 24: 1460-1466. Streptomyces is a Gram-positive bacterium with the potential a produce beneficial antimicrobials. This study aimed to determine the antimicrobial activity, morphological characteristics, cytotoxicity, and chemical profile of S. koyangensis SHP 9-3. This bacterium was isolated from Enggano Island, Indonesia, and the morphological characteristics were determined using a Scanning Electron Microscope (SEM). Cultivation was conducted in the International Streptomyces Project (ISP-2) media using baffled flasks and a shaker incubator. The secondary metabolites were conducted on the extract against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans by a twofold dilution method. Concurrently, cytotoxicity testing was done on Vero cells. The extract's chemical profile was analyzed using LC-MS/MS. The results showed a prominent antibacterial activity as the extract inhibited the growth of S. aureus and C. albicans at 7.81 µg/mL. However, a growth inhibition against E. coli was observed at 250 µg/mL and P. aeruginosa at 31.25 µg/mL. The cytotoxic test showed the IC50 value on Vero cells was 5,015 µg/mL, indicating that the extract was nontoxic against Vero cells. The chemical profile of the extract by LC-MS/MS revealed the presence of five major components, i.e., cyclo (Pro-Val) (C10H16N2O2), kuraramine (C12H18N2O2), 1,4-diphenyl-1-pentanone (C17H18O), (3S,4S)-4-amino-3-hydroxy-6-methyl heptanoic acid (C21H42N2O7), and 2-ethyl-8-methyl-2,8-diazaspiro (4,5) decane-1,3-dione (C11H18N2O2).

Cacao is attractive for its flavor and taste and its beneficial effects. Therefore, this commodity is promising to be developed as a functional food. Many studies reported the bioactive compounds in cacao beans and their health benefits. However, to attain desirable flavor and taste, the curing process of cacao beans is a common practice, in which fermentation is one of the processes. Despite its significance, fermentation also alters the bioactive components in cacao beans. To develop a functional food rich in bioactive compounds, measuring the bioactive contents of treated and untreated cacao beans is desired. For that reason, in this study, the analysis of total polyphenol, flavonoid, catechin, and epicatechin, as well as the radical scavenging activity of fermented cacao beans, was performed. The study revealed that fermentation negatively affected all parameters. Cacao beans fermentation up to 96 h resulted in the loss of 54% of total polyphenol, 77% of total flavonoid, and 56% of radical scavenging activity. Determination of the flavan-3-ol components, catechin, and epicatechin, revealed that they were reduced as the fermentation occurred.