Day :
- Applied Microbiology
Chair
Dewinta Enggar Pramesthi
Airlangga University, Indonesia
Co-Chair
Manik R Wahyunitisari
Airlangga University, Indonesia
Session Introduction
Learn-Han Lee
Monash University, Malaysia.
Title: Discovery of anti-MRSA protein from mangrove-derived Streptomyces pluripotens MUSC 135T
Time : 12:10-12:50
Biography:
The emergence of multidrug resistant bacteria or “super bug” has called on the scientific community to search for more potent, effective drugs to keep these pathogens under control. At the forefront of the production of bioactive metabolites, microorganisms have been recognized as “mini-factories” which are capable of synthesizing interesting bioactive natural compounds with reasonable cost. Under the family of Actinobacteria, the genus Streptomyces stands out in terms of manufacturing bioactive metabolites reserves, contributing over 10,000 bioactive compounds with high pharmaceutical values. In fact, the discovery of antibiotic streptomycin from soil bacterium, Streptomyces griseus has bestowed Professor Waksman and his team with the award of the Nobel Prize in Physiology or Medicine in 1952. In continuing the fight against deadly infections, the search novel Streptomyces-derived bioactive compound is further bolstered with the numerous beneficial implications behind. The mangrove forest is termed as the intertidal region located between the land and the sea. Despite the constant exposure to harsh conditions, some Actinobacteria isolated from these ecosystems have exhibited interesting bioactivities, including anti-MRSA, antifungal and so on. Streptomyces pluripotens MUSC 135T was isolated as novel species from the poorly explored mangrove sediment (East Coast, Peninsular Malaysia). As an attempt to study the bioactive potential of this strain, MUSC 135T was subjected to fermentation before conducting antibacterial assays using traditional agar well diffusion method and high throughput screening method with 96-well microplates. Intriguely, the inhibition zone of MUSC 135T crude extract against MRSA (10.5 mm) was larger than that of the positive control, a Vancomycin disc (30 µg), which exhibited an inhibition zone of 9 mm. A deeper investigation using next generation sequencing has revealed some fascinating bioactive potential of strain MUSC 135T (NCBI accession: CP021080.1); a total of 4 biosynthetic gene clusters related to production of bacteriocin has been identified from MUSC 135T with genome size of 7.34 Mbps using bioinformatics tool BAGEL4. Among these cluster, one of them was predicted to be responsible for the production of Linear Azol(in)e-containing Peptides (LAPs), a group of bioactive metabolites which have been reported for anti-MRSA activities. Subsequently, one of the predicted genes was selected for expression study using E. coli model and the purified protein (Protein135_#1) was then used for anti-MRSA screening. Preliminary anti-MRSA screening has revealed significant reduction of MRSA after the treatment of Protein135_#1. In conclusion, these findings highlight the importance in novel strain from underexplored area, like mangrove forest, particularly in the search of useful bioactive compounds.
Abstract:
Learn-Han Lee has completed his PhD in Molecular Biology and currently working as the Head of Microbiome and Bio-resource Research Strength, Principle Investigator of Novel Bacteria and Drug Discovery Research Group, Monash University Malaysia. He is the Lifetime Member of Bergey’s International Society for Microbial Systematics (BISMiS) and a Member of Royal Society of Biology (MRSB, UK).
Manik R Wahyunitisari
Airlangga University, Indonesia
Title: Crosstalk between cholecalciferol, retinyl palmitate and tuberculosis
Time : 13:50-14:30
Biography:
Manik R Wahyunitisari has completed her Medical Doctor from Airlangga University Faculty of Medicine, Indonesia
Abstract:
Vitamin A and vitamin D contribute on Tuberculosis (TB) pathogenesis. The purpose of this study is to analyze are the combination of cholecalciferol and retinyl palmitate could induce the effectiveness of 2nd line anti-TB drugs. Mus musculus C3HeB/FeJ was infected by multidrug-resistant strain Mtb. The first group (G1) was euthanized two weeks after infected to evaluate lung TB forming. (G2) is control group without any therapy. (G3) was given 2nd line anti-TB drugs. (G4) was given retinyl palmitate and 2nd line anti-TB drugs. (G5) was given 2nd line anti-TB drugs and cholecalciferol. (G6) was given 2nd line anti-TB drugs, retinyl palmitate and cholecalciferol. Immunohistochemistry was used for measuring nuclear receptor expression of vitamin D (VDR) and vitamin A (RARγ2); apoptosis caspase-3 marker; autophagy markers CRAMP and LC3B; necrosis marker RIPK3; and interstitial collagenase MMP1. Bacteria viability was counted in Colony Forming Units (CFU). Partial least square structural equation modeling (PLS-SEM) was used to analyze structural model within variable. VDR and RARγ2 were increased by each ligand (p=0.026 and p=0.019). Cholecalciferol increases autophagy which is characterized by an increase in CRAMP and LC3B (p=0.000 and p=0.001) and decreases MMP1 (p=0.010). Cholecalciferol or retinyl palmitate both increased casp3 expression (p=0.035 and p=0.027) and both reduced CFU (p=0.000 and p=0.000). The combination of cholecalciferol and retinyl palmitate reduced RIPK3 (p=0.002). Our study proves that the combination of cholecalciferol and retinyl palmitate supplementation on the 2nd line anti-TB drugs reduces cell necrosis directly.
Dewinta Enggar Pramesthi
Airlangga University, Indonesia
Title: Crosstalk between cholecalciferol, retinyl palmitate and tuberculosis
Time : 15:10-15:50
Biography:
Dewinta Enggar Pramesthi has completed her Medical Doctor from Airlangga University Faculty of Medicine, Indonesia.
Abstract:
Vitamin A and vitamin D contribute on Tuberculosis (TB) pathogenesis. The purpose of this study is to analyze are the combination of cholecalciferol and retinyl palmitate could induce the effectiveness of 2nd line anti-TB drugs. Mus musculus C3HeB/FeJ was infected by multidrug-resistant strain Mtb. The first group (G1) was euthanized two weeks after infected to evaluate lung TB forming. (G2) is control group without any therapy. (G3) was given 2nd line anti-TB drugs. (G4) was given retinyl palmitate and 2nd line anti-TB drugs. (G5) was given 2nd line anti-TB drugs and cholecalciferol. (G6) was given 2nd line anti-TB drugs, retinyl palmitate and cholecalciferol. Immunohistochemistry was used for measuring nuclear receptor expression of vitamin D (VDR) and vitamin A (RARγ2); apoptosis caspase-3 marker; autophagy markers CRAMP and LC3B; necrosis marker RIPK3; and interstitial collagenase MMP1. Bacteria viability was counted in Colony Forming Units (CFU). Partial least square structural equation modeling (PLS-SEM) was used to analyze structural model within variable. VDR and RARγ2 were increased by each ligand (p=0.026 and p=0.019). Cholecalciferol increases autophagy which is characterized by an increase in CRAMP and LC3B (p=0.000 and p=0.001) and decreases MMP1 (p=0.010). Cholecalciferol or retinyl palmitate both increased casp3 expression (p=0.035 and p=0.027) and both reduced CFU (p=0.000 and p=0.000). The combination of cholecalciferol and retinyl palmitate reduced RIPK3 (p=0.002). Our study proves that the combination of cholecalciferol and retinyl palmitate supplementation on the 2nd line anti-TB drugs reduces cell necrosis directly.