HAYATI Journal of Biosciences
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Identification, Characterization and Antioxidant Activity of Yellowish-Orange Pigments Actinobacteria
Get PDFActinobacteria are Gram-positive bacteria that can produce various secondary metabolites, including pigments that have potential as antioxidants. This study assessed the characteristics of yellowish-orange pigmented actinobacteria. The antioxidant activity of their extract pigment was tested by DPPH and analyzed by LC-MS. The identification results based on the 16S rRNA gene showed that the HRA isolate had 99.91% similarity value to Streptomyces gramineus JR-43, the HVA isolate had 99.22% similarities value to S. tendae ATCC 19812 and the HVB isolate had 81.18% similarities value to Rhodococcus ruber DSM 43338. Meanwhile, AGM 2.2 and 2.3 isolates had similarities to S. xanthophaeus NRRL B-5414 with similarity values of 98.95% and 99.82%, respectively. The Inhibitory Concentration (IC50) of actinobacterial crude extract pigments ranges from 53.38 µg/ml to 184.38 µg/ml. The HVB isolates with the major compound luteolin, have the highest antioxidant activity. The crude extract pigment of HVB isolates may consider rich in luteolin and has potency as an antioxidant
Diversity of Harvested Gastropods in Guang-Guang, Mati City, Davao Oriental, Philippines
Get PDFGastropods are a highly abundant group of mollusks in the marine environment. Marine gastropods primarily function as prey for other animals, grazers which help recycle nutrients and increase bioturbation activities in the marine ecosystem. Edible gastropods are significant to the economy of coastal communities. However, overexploitation leads to harming the population of marine gastropods. The study aimed to determine the diversity of gastropods harvested at Guang-guang, Mati City, Davao Oriental. It also assessed the abundance of the identified gastropods in the area. This study used purposive sampling to enroll gleaners (N = 30) who collected shells and monitored their gastropod harvest. The harvested gastropods were segregated, counted, and identified with the help of gleaners. A total of ten gastropod species were identified from the family of Strombidae, Conidae, Neritidae, Cypraeidae, and Turbinidae. The most abundant group after harvest was Canarium urceus, with a relative abundance of 67%, followed by Ilyanassa obsoleta, with 22%. However, the least harvested species was Nerita, with a relative abundance of 0.18%. The findings revealed that the harvested gastropods in Guang-guang have a diversity index of 0.99, indicating a low diversity of gastropods in the area. This low diversity could mean the area experienced overexploitation through time as gleaners have heavily harvested gastropods. With that, it is highly recommended that conservation should be prioritized to preserve the seashells
The Toxicity Test of Synthetic Insecticides on Tetragonula laeviceps (Apidae: Meliponini)
Get PDFIntense insecticide application is thought to cause a decline in bee colonies worldwide. Bees are effective pollinators in increasing the production of agricultural commodities. The stingless bee Tetragonula laeviceps (Apidae: Meliponini) is widely cultivated and found around plantations. The insecticides used in the toxicity test were imidacloprid 200 g/L, fipronil 50 g/L, lambda-cyhalothrin 25 g/L, profenofos 500 g/L, and chlorantraniliprole 50 g/L followed by a semi-field test using imidacloprid, fipronil, and profenofos insecticides on the cucumber plant. The LC50 value showed that exposure to imidacloprid, fipronil, and profenofos insecticides caused toxic effects on T. laeviceps by contact and orally. Lambda-cyhalothrin was found harmful on contact exposure, in contrast, chlorantraniliprole was harmful through oral. Classification of insecticide toxicity based on LD50 contact for imidacloprid, fipronil, lambda-cyhalothrin, and profenofos were very toxic and needed a risk assessment. However, chlorantraniliprole was classified as moderately toxic and low risk. In semi-field test results, imidacloprid and fipronil insecticides caused a significant decrease in the leave-return and bee-visiting activity on cucumber flowers. These insecticides also produced a low average yield of fruit weight. Insecticide application can affect the role of T. laeviceps as the pollinator which impacts the production of agricultural commodities
Cloning and Extracellular Expression of Glargine in Pichia pastoris
Get PDFPatients with diabetes mellitus increase significantly every year. The increasing number of people with diabetes mellitus results in increased insulin requirements. There are two types of insulin used for diabetes mellitus treatment: human insulin and insulin analogues. Escherichia coli, Pichia pastoris, Saccharomyces cerevisiae, or Hansenula polymorpaha has been used to produce human insulin and insulin analogues. Pichia pastoris can produce glargine in large quantities, and the insulin protein produced will be secreted outside the cell to facilitate the purification process. The advantage of glargine has a long working time of up to 24 hours. Hence, glargine is more effective because patients with diabetes receive glargine injections only once daily. The research started with cloning the glargine gene, transforming pPICZαA-G plasmid into Pichia pastoris, and testing glargine production. 20 recombinant Pichia pastoris colonies were selected and regenerated. Eight recombinant Pichia pastoris colonies were tested for glargine production, and six colonies were detected producing glargine by electrophoresis SDS-PAGE gel stained with Coomassie blue. This study aims to produce glargine using Pichia pastoris as an expression system capable of producing glargine extracellularly, thus simplifying the purification process
Phytotoxic and Cytotoxic Polyketides Produced by Fungal Endophytes Isolated from Psidium guajava
Get PDFEndophytic fungi reside within their host plants with no obvious symptoms. They have been proven as abundant sources of new bioactive compounds with diverse structures. In our continuous search of the new compound from endophytic fungi, a new compound, 5,7-dihydroxy-3(R)-methylphthalide (1), was isolated from Xylaria brevipes PGR1, and three known compounds, (–)-altenuene (2), alternariol (3), and altertoxin I (4), were isolated from Alternaria alternata D-8. Both endophytic fungi were isolated from the branch of an Indonesian medicinal plant, Psidium guajava. Their structures were elucidated using extensive spectroscopy analyses, including 1D/2D NMR, HRESITOFMS, and data comparisons with the literature. Compounds 1–3 exhibited phytotoxicity at IC50 (µg/ml) values of 4.6, 23.07, and 77.28, respectively. Additionally, 1–4 showed significant cytotoxicity in the brine shrimp lethality test (BSLT), with mortality rates at 75%, 50%, 90%, and 80% at the concentration of 0.19 μg/ml, respectively
Expression of SARS-CoV-2 Nucleocapsid (N) Recombinant Protein Using Escherichia coli System
Get PDFOne of the main antigen that can be used for serological testing is the nucleocapsid (N) which is the most abundant viral-derived protein in SARS-CoV-2 where this virus can cause COVID19 disease. The aim of this study was to develop the SARS-CoV-2 N recombinant protein using Escherichia coli expression system. A total of 1,089 nucleotides encoding 362 amino acids of SARS-CoV-2 N was cloned to pET-14b vector. The plasmid then expressed in E. coli BL21 (DE3) and induced with 1.0 mM IPTG (Isopropyl-β-d-1-thiogalactopyranoside). The cell was harvested using denaturation lysis buffer due to inclusion body formation of SARS-CoV-2 N protein. Dialysis processed and concentrated using PEG-6000 resulted 0.992 mg/ml protein yield. Analysis of SARS-CoV-2 N recombinant protein using SDS-PAGE technique showed approximately 37.0 kDa specific band target protein. Application of this SARS-CoV-2 N recombinant protein to vaccinated and non-vaccinated antibody serum samples using ELISA technique indicated the significant result of optical density mean at 0.603 and 0.135, respectively. This study revealed that the production of SARS-COV-2 N recombinant protein could be carried out in E. coli expression system under denatured conditions, therefore the methods are more effective in producing the protein as a basic material in immuno-diagnostic assay
Phytoremediation of Acid Mine Drainage with Melaleuca cajuputi, Nauclea orientalis, and Vetiveria zizanioides in Floating Treatment Wetland
Get PDFThe formation of acid mine drainage (AMD) is a common environmental problem in the mining industry. Its passive management through wetland construction has gained more consideration in recent years. However, the application in the field is constrained by the large area and relatively shallow depth. Indonesia has no passive technology to neutralize AMD in deep water. One solution is to apply a floating treatment wetland (FTW) system. Therefore, this study aimed to determine the ability of several hyperaccumulator plants, such as Melaleuca cajuputi, Nauclea orientalis, and Vetiveria zizanioides, to neutralize AMD using a floating system by conducting FTW trials. The method used was a Completely Randomized Design (CRD) with 3 treatments and control/without plants. Each treatment had 3 replications, resulting in 12 experimental units. The results showed that the FTW with or without plants could increase pH and decrease dissolved Mn by 75.31-90.74%. Heavy metals were chelated by organic matter, absorbed by plants, and deposited in the form of metal sulfides. The results also indicated that besides having a positive effect on pH and heavy metal reduction, the organic-based floating wetland increased biological oxygen demand (BOD) from 61.08-79.71%
Evaluation of the Community Structure Leafminer Fly, Liriomyza spp. (Diptera: Agromyzidae) and Their Parasitoids on Various Host Plant Families in Bali Province
Get PDFLeaf miners, Liriomyza spp., have developed as a severe pest in recent years, reducing the production of several ornamental and crops in Indonesia. As a result, monitoring and control mechanisms have been proposed. This research aimed to investigate community structure and identify Liriomyza spp., which impacts crops and natural parasitoids in Bali Province. Leaf samples of vegetables, ornamental plants, and related species were collected in all regencies/cities of Bali Province in 2019 and 2020 and preserved in the laboratory for observation and counting of leaf miners and related parasitoids. The findings revealed that four Liriomyza spp. were detected in host plants, namely Liriomyza sativae Blanchard (Diptera: Agromyzidae), Liriomyza huidobrensis Blanchard (Diptera: Agromyzidae), Liriomyza chinensis Kato (Diptera: Agromyzidae), and Liriomyza trifolii Burgess (Diptera: Agromyzidae). Asteraceae species were the most common host plants attacked by leaf miners in the field. The same occurrence was discovered at both heights (high and lowlands). The dry season is the most crucial component in the abundance of Liriomyza spp. In addition, parasitoids of the type Opius cromatomiyae Belokobylskij and Wharton (Hymenoptera: Braconidae) and Hemiptarsenun varicornis Girault (Hymenoptera: Eulophidae) were identified in abundance in Asteraceae plants. Similarly, the parasitization rate of the two parasitoids was found to be high in plants of the Asteraceae and Brassicae families. This approach highlights the decision-making process for controlling Liriomyza spp. on diverse host plant families by developing ecologically beneficial and sustainable parasitoids
An Early Reference to DNA Barcode for the Anacardiaceae Family
Get PDFAnacardiaceae is well-known for its edible fruits and economically important species in Indonesia. Approximately 3% of Indonesia\u27s endangered and vulnerable species belong to this family. Fast and accurate species identification is crucial to support the conservation efforts for this family, such as employing DNA sequences. Species identification using DNA sequences, known as DNA barcoding, has been widely used in many applied fields. So far, the application of DNA barcoding for Anacardiaceae plant species is limited to several genera only, such as Mangifera and Spondias. This research aimed to enrich the DNA barcode references of Anacardiaceae and to evaluate the most suitable and effective genetic marker as DNA barcodes to identify species of 35 samples representing sixteen species of the Anacardiaceae family using chloroplast markers matK and rbcL as barcode regions. When comparing the morphological identification with the molecular assignments, the barcode accuracy was 62.50% (matK), 60.61% (rbcL), and 73.33% (matK+rbcL) at the genus level. All the markers failed to show a barcoding gap, even though the t-test showed that the intraspecific and interspecific genetic distances were significantly different for matK and rbcL+matK. Among others, Gluta walichii (Hook.f.) Ding Hou and Melanochyla caesia Jack were the only species successfully resolved by all markers. Nevertheless, new DNA barcodes of six Anacardiaceae species were made available by this study, enriching the genetic references of tropical flora diversity
Red Betel Leaf Bioactive Compounds as ERα Receptor Inhibitors In Silico and MCF-7 Cell Anticancer In Vitro
Get PDFCancer is one of the leading causes of death in the world. Excess endogenous estrogen is a risk factor for breast cancer. Red betel leaf herbal plants have been used as an alternative cervical and colon cancer treatment. This study aimed to obtain the active compounds that play a role in ERα receptor inhibitors in silico and to determine the anti-breast cancer cytotoxic activity of the extract and fraction of red betel leaf against MCF-7 cells. We use in silico research method using the YASARA Structure software with anti-breast cancer receptors, namely 3ERT, and in vitro using the MTT test on the anti-breast cancer cytotoxic activity of MCF-7 cells. There are 38 compounds that were obtained from the research. The results of the in silico test showed that the bioactive compound that played a role in inhibiting the ERα was 2-(4-Hydro xyphenyl)-2-phenyl-N(3,3-diphenylpropyl)-acetamide (44578655), a compound from water fraction, with an inhibition constant of 2.82 × 10-8 µM and Gibbs free energy of –10.2880 Kcal/mol. In vitro results showed that the best cell growth inhibition value was obtained from the n-hexane fraction at a concentration of 500 ppm of 73.42%. The conclusions of this study indicate that the bioactive compound of red betel leaf is water fraction is the best fraction inhibition. However, the hexane fraction proved to have cytotoxic activity against breast cancer MCF-7 cells