Hyun Park

Low-temperature growth of layered molybdenum disulphide with controlled clusters

Layered molybdenum disulphide was grown at a low-temperature of 350u2009°C using chemical vapour deposition by elaborately controlling the cluster size. The molybdenum disulphide grown under various sulphur-reaction-gas to molybdenum-precursor partial-pressure ratios were examined. Using spectroscopy and microscopy, the effect of the cluster size on the layered growth was investigated in terms of the morphology, grain size, and impurity incorporation. Triangular single-crystal domains were grown at an optimized sulphur-reaction-gas to molybdenum-precursor partial-pressure ratio. Furthermore, it is proved that the nucleation sites on the silicon-dioxide substrate were related with the grain size. A polycrystalline monolayer with the 100-nm grain size was grown on a nucleation site confined substrate by high-vacuum annealing. In addition, a field-effect transistor was fabricated with a MoS2 monolayer and exhibited a mobility and on/off ratio of 0.15u2009cm2 V−1 s−1 and 105, respectively.

Proteomic and transcriptomic investigations on cold-responsive properties of the psychrophilic Antarctic bacterium Psychrobacter sp. PAMC 21119 at subzero temperatures.

Psychrobacter sp. PAMC 21119, isolated from Antarctic permafrost soil, grows and proliferates at subzero temperatures. However, its major mechanism of cold adaptation regulation remains poorly understood. We investigated the transcriptomic and proteomic responses of this species to cold temperatures by comparing profiles at -5°C and 20°C to understand how extreme microorganisms survive under subzero conditions. We found a total of 2,906 transcripts and 584 differentially expressed genes (≥ twofold, P <0.005) by RNA-seq. Genes for translation, ribosomal structure and biogenesis were upregulated, and lipid transport and metabolism was downregulated at low temperatures. A total of 60 protein spots (≥ 1.8 fold, Pu2009<u20090.005) showed differential expression on two-dimensional gel electrophoresis and the proteins were identified by mass spectrometry. The most prominent upregulated proteins in response to cold were involved in metabolite transport, protein folding and membrane fluidity. Proteins involved in energy production and conversion, and heme protein synthesis were downregulated. Moreover, isoform exchange of cold-shock proteins was detected at both temperatures. Interestingly, pathways for acetyl-CoA metabolism, putrescine synthesis and amino acid metabolism were upregulated. This study highlights some of the strategies and different physiological states that Psychrobacter sp. PAMC 21119 has developed to adapt to the cold environment in Antarctica.

A transcriptomic analysis of the response of the arctic pteropod Limacina helicina to carbon dioxide-driven seawater acidification

Ocean acidification from the uptake of anthropogenic carbon dioxide (CO2) is regarded as a critical threat particularly to marine calcifying organisms. The arctic pteropod Limacina helicina may be one of the first polar organisms that are expected to display early sensitivity to ocean acidification, but a molecular approach as a foundation for understanding the effect of ocean acidification on this pteropod has rarely been reported. In this study, we examined the sublethal effects of CO2-driven seawater acidification at the transcriptome level in L. helicina. cDNAs, treated under control (pH 8.2), high-CO2 (pH 7.5), and extreme-CO2 (pH 6.5) conditions, generated a total of 31,999,474 reads, comprising a total of 2,271,962,654xa0bp, using the Illumina platform. De novo assembly yielded 53,121 transcripts comprising 31.79xa0Mbp. Among the upregulated genes, 346 (0.7xa0%) and 655 (1.2xa0%) genes responded to extreme-level CO2 (pH 6.5) and high-level CO2 (pH 7.5), respectively. Also, 76 (0.1xa0%) transcripts were commonly upregulated in both conditions. Among the downregulated genes, 690 (1.3xa0%) and 739 (1.4xa0%) genes were in response to extreme-level CO2 and high-level CO2, respectively. Also, 270 downregulated genes (0.5xa0%) were affected in both acidic stress conditions. Moreover, 504 transcripts (1xa0%) of biomineralization-related genes were identified; 16 of these genes showed differential expression in response to acidified seawater. The dataset provides the first comprehensive overview of changes in transcript levels in the arctic pteropod L. helicina in response to increased CO2, emphasizing the potential impact of future environmental change and ocean acidification on Arctic species with external calcified structures.

Effect of the Antifreeze Protein from the Arctic Yeast Leucosporidium sp. AY30 on Cryopreservation of the Marine Diatom Phaeodactylum tricornutum

Antifreeze proteins are a group of proteins that allow organisms to survive in subzero environments. These proteins possess thermal hysteresis and ice recrystallization inhibition activities. In the present study, we demonstrated the efficiency of a recombinant antifreeze protein from the Arctic yeast Leucosporidium sp. AY30, LeIBP, in cryopreservation of the marine diatom Phaeodactylum tricornutum, which is one of the classical model diatoms and has most widely been studied with regard to its ecology, physiology, biochemistry, and molecular biology. P. tricornutum cells were frozen by either a fast or two-step freezing method in freezing medium containing 10xa0% dimethyl sulfoxide, glycerol, propylene glycol, and ethylene glycol, respectively, with or without LeIBP supplement. When cells were frozen using the two-step freezing method, cell survival was significantly increased and statistically the same as that of unfrozen native cells in the presence of 0.1xa0mg/ml LeIBP in 10xa0% propylene glycol or 10xa0% ethylene glycol at day 11 of post-thaw culture. In the presence of LeIBP, the concentration of chlorophyll a was dramatically increased to 14-, 48-, 1.6-, and 8.8-fold when cells were frozen in freezing medium containing dimethyl sulfoxide (DMSO), glycerol, propylene glycol (PG), and ethylene glycol (EG), respectively. Scanning electron microscopy observations demonstrated that the cells were also successfully preserved and epitheca or hypotheca were not deformed. These results demonstrate that LeIBP was successfully applied to improve cryopreservation of the marine diatom P. tricornutum.

One-step large-scale synthesis of micrometer-sized silver nanosheets by a template-free electrochemical method

We have synthesized micrometer-sized Ag nanosheets via a facile, one-step, template-free electrochemical deposition in an ultra-dilute silver nitrate aqueous electrolyte. The nanosheet growth was revealed to occur in three stages: (1) formation of polygonal Ag nuclei on a substrate, (2) growth of {112}-faceted nanowire from the nuclei, and (3) anisotropic growth of (111)-planar nanosheets, approximately 20 to 50 nm in thickness and 10 μm in width, in the <112>−direction. The vertical growth of the facet nanowire was induced by the strong interface anisotropy between the deposit and electrolyte due to the ultra-dilute concentration of electrolyte and high reduction potential. The thickness of Ag nanosheets was controllable by the adjustment of the reduction/oxidation potential and frequency of the reverse-pulse potentiodynamic mode.

Structural basis for the ligand-binding specificity of fatty acid-binding proteins (pFABP4 and pFABP5) in gentoo penguin

Fatty acid-binding proteins (FABPs) are involved in transporting hydrophobic fatty acids between various aqueous compartments of the cell by directly binding ligands inside their β-barrel cavities. Here, we report the crystal structures of ligand-unbound pFABP4, linoleate-bound pFABP4, and palmitate-bound pFABP5, obtained from gentoo penguin (Pygoscelis papua), at a resolution of 2.1xa0Å, 2.2xa0Å, and 2.3xa0Å, respectively. The pFABP4 and pFABP5 proteins have a canonical β-barrel structure with two short α-helices that form a cap region and fatty acid ligand binding sites in the hydrophobic cavity within the β-barrel structure. Linoleate-bound pFABP4 and palmitate-bound pFABP5 possess different ligand-binding modes and a unique ligand-binding pocket due to several sequence dissimilarities (A76/L78, T30/M32, underlining indicates pFABP4 residues) between the two proteins. Structural comparison revealed significantly different conformational changes in the β3-β4 loop region (residues 57-62) as well as the flipped Phe60 residue of pFABP5 than that in pFABP4 (the corresponding residue is Phe58). A ligand-binding study using fluorophore displacement assays shows that pFABP4 has a relatively strong affinity for linoleate as compared to pFABP5. In contrast, pFABP5 exhibits higher affinity for palmitate than that for pFABP4. In conclusion, our high-resolution structures and ligand-binding studies provide useful insights into the ligand-binding preferences of pFABPs based on key protein-ligand interactions.

Crystal Structure and Functional Characterization of an Esterase (EaEST) from Exiguobacterium antarcticum.

A novel microbial esterase, EaEST, from a psychrophilic bacterium Exiguobacterium antarcticum B7, was identified and characterized. To our knowledge, this is the first report describing structural analysis and biochemical characterization of an esterase isolated from the genus Exiguobacterium. Crystal structure of EaEST, determined at a resolution of 1.9 Å, showed that the enzyme has a canonical α/β hydrolase fold with an α-helical cap domain and a catalytic triad consisting of Ser96, Asp220, and His248. Interestingly, the active site of the structure of EaEST is occupied by a peracetate molecule, which is the product of perhydrolysis of acetate. This result suggests that EaEST may have perhydrolase activity. The activity assay showed that EaEST has significant perhydrolase and esterase activity with respect to short-chain p-nitrophenyl esters (≤C8), naphthyl derivatives, phenyl acetate, and glyceryl tributyrate. However, the S96A single mutant had low esterase and perhydrolase activity. Moreover, the L27A mutant showed low levels of protein expression and solubility as well as preference for different substrates. On conducting an enantioselectivity analysis using R- and S-methyl-3-hydroxy-2-methylpropionate, a preference for R-enantiomers was observed. Surprisingly, immobilized EaEST was found to not only retain 200% of its initial activity after incubation for 1 h at 80°C, but also retained more than 60% of its initial activity after 20 cycles of reutilization. This research will serve as basis for future engineering of this esterase for biotechnological and industrial applications.

The complete chloroplast genome of Antarctic pearlwort, Colobanthus quitensis (Kunth) Bartl. (Caryophyllaceae)

Abstract We determined the complete chloroplast DNA sequence of an extremophile plant, Colobanthus quitensis (Antarctic pearlwort), by de novo assembly based on the sequencing results from Illumina MiSeq platform (Illumina Inc., San Diego, CA). The chloroplast genome of C. quitensis (NCBI accession no. KT737383) is a sequence of 151 276u2009bp long with a typical quadripartite structure composed of a large single copy region, a small single copy region and a pair of inverted repeats. The overall GC content of C. quitensis genome is 36.7% and it has 66 simple sequence repeats. It contains a total 112 genes including 78 protein coding genes, 30 tRNA genes, and four rRNA genes.

Complete mitochondrial genome of the Antarctic bullhead notothen, Notothenia coriiceps (Perciformes, Nototheniidae)

Abstract The complete sequence of the mitochondrial genome of Notothenia coriiceps was obtained by genome assembly. The complete sequence was determined to be 18,347 base pairs in length and to contain 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes, and 2 control regions. Of the thirteen protein-coding genes, two genes (cox1 and atp6) had GTG start codons, and six genes (nad2, cox2, cox3, nad3, nad4, and cytb) had incomplete stop codons that require the post-transcriptional addition of A bases. The base composition of the mitogenome was 26.3% A, 27.6% T, 17.5% G, and 28.5% C.

Crystal structure and modeling of the tetrahedral intermediate state of methylmalonate-semialdehyde dehydrogenase (MMSDH) from Oceanimonas doudoroffii

The gene product of dddC (Uniprot code G5CZI2), from the Gram-negative marine bacterium Oceanimonas doudoroffii, is a methylmalonate-semialdehyde dehydrogenase (OdoMMSDH) enzyme. MMSDH is a member of the aldehyde dehydrogenase superfamily, and it catalyzes the NADdependent decarboxylation of methylmalonate semialdehyde to propionyl-CoA. We determined the crystal structure of OdoMMSDH at 2.9 Å resolution. Among the twelve molecules in the asymmetric unit, six subunits complexed with NAD, which was carried along the protein purification steps. OdoMMSDH exists as a stable homodimer in solution; each subunit consists of three distinct domains: an NAD-binding domain, a catalytic domain, and an oligomerization domain. Computational modeling studies of the OdoMMSDH structure revealed key residues important for substrate recognition and tetrahedral intermediate stabilization. Two basic residues (Arg103 and Arg279) and six hydrophobic residues (Phe150, Met153, Val154, Trp157, Met281, and Phe449) were found to be important for tetrahedral intermediate binding. Modeling data also suggested that the backbone amide of Cys280 and the side chain amine of Asn149 function as the oxyanion hole during the enzymatic reaction. Our results provide useful insights into the substrate recognition site residues and catalytic mechanism of OdoMMSDH.