12^th Annual Meet on Bacteriology & Applied Microbiology September 23-24, 2019 Tokyo, Japan

Hara Jang has her Expertise in Protein Purification and Crystallization. She has completed her Bachelor’s degree in Biochemistry at Chonbuk National University and also PhD from University of Maryland.

Concanavalin A (ConA) interacts with carbohydrates as a lectin, and recent reports proposed its application for detecting a diversity of viruses and pathogens. Structural studies have detailed the interaction between ConA and carbohydrates and the metal coordination environment with manganese and calcium ions (Mn-Ca-ConA). ConA was crystallized with a cadmium-containing precipitant, and the refined structure indicates that Mn was replaced by Cd (Cd-Ca-ConA). The structural comparison with ConA demonstrates that the metal-coordinated residues of Cd-Ca-ConA, that is Glu8, Asp10, Asn14, Asp19, and His24, do not have conformational shifts, but residues for sugar binding, including Arg228, Tyr100, and Leu99, reorient their side chains, slightly. Previous studies demonstrated that excess cadmium ions can coordinate with other residues, including Glu87 and Glu183, which were not coordinated with Cd in this study. The trimeric ConA in this study coordinated Cd with other residues, including Asp80 and Asp82, for complex generation. The monomer does not have specific interaction near interface regions with the other monomer, but secondary cadmium coordinated with two aspartates (Asp80 and Asp82) from monomer 1 and one aspartate (Asp16) from monomer 2. This study demonstrated that complex generation was induced via coordination with secondary Cd and showed the application potential regarding the design of complex formation for specific interactions with target saccharides.

Heeseon Yoo has her Expertise in Characterization of Metallo-Proteins. She has completed her BA in Chemistry and Biological Chemistry Lab at Chonbuk National University and also PhD from University of Maryland

Hydroxylation through Soluble Methane Monooxygenase (sMMO) has been studied extensively to understand the biological conversion from methane to methanol in ambient conditions, although more detailed mechanisms of this biocatalyst are yet undiscovered. In this study, sMMO components, including the hydroxylase (MMOH), regulatory (MMOB), and reductase (MMOR), were expressed and purified from a type-II methanotroph, Methylosinus sporium strain 5 (M. sporium 5), to characterize its hydroxylation mechanism. Brown–black pigments were detected in Ms. sporium 5 during cell growth, with the optical density value of 8.0 at 600 nm (OD600), and this change was dependent on the ratio of cell: iron. The cell extract showed that MMOH, MMOB, and MMOR were expressed by M. sporium 5, and these enzymes were extracted with high purity. The enzyme activity confirmed that 2.0 mol equivalents of MMOB is necessary to achieve catalytic activities, and sMMO from M. sporium 5 oxidized a broad range of substrates, including alkanes, alkenes, halogens and aromatics. Optimal activities were observed at pH 7.5 in most substrates, possibly because of the electron transfer environment in MMOR, and this was explained by the optical spectra from FAD-containing domains

Ka Young Son has her Expertise in purification and characterization of redox enzymes. She has completed her BA and MS at Chonbuk National University and also PhD from University of Maryland.

Methane hydroxylation through Methane Monooxygenases (MMOs) is a key aspect due to their control of the carbon cycle in the ecology system and recent applications of methane gas in the field of bioenergy and bioremediation. Methanotropic bacteria perform a specific microbial conversion from methane, one of the most stable carbon compounds, to methanol through elaborate mechanisms. MMOs express Particulate Methane Monooxygenase (pMMO) in most strains and Soluble Methane Monooxygenase (sMMO) under copper-limited conditions. The mechanisms of MMO have been widely studied from sMMO belonging to the Bacterial Multicomponent Monooxygenase (BMM) superfamily. This enzyme has diiron active sites where different types of hydrocarbons are oxidized through orchestrated hydroxylase, regulatory and reductase components for precise control of hydrocarbons, oxygen, protons and electrons. Recent advances in biophysical studies, including structural and enzymatic achievements for sMMO, have explained component interactions, substrate pathways, and intermediates of sMMO. In this account, oxidation of methane in sMMO is discussed with recent progress that is critical for understanding the microbial applications of C-H activation in one-carbon substrates.

Dewinta Enggar Pramesthi has completed her Medical Doctor from Airlangga University Faculty of Medicine, Indonesia

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