The Versatile Manipulations of Self-Assembled Proteins in Vaccine Design

Protein assemblies provide unique structural features which make them useful as carrier molecules in biomedical and chemical science. Protein assemblies can accommodate a variety of organic, inorganic and biological molecules such as small proteins and peptides and have been used in development of subunit vaccines via display parts of viral pathogens or antigens. Such subunit vaccines are much safer than traditional vaccines based on inactivated pathogens which are more likely to produce side-effects. Therefore, to tackle a pandemic and rapidly produce safer and more effective subunit vaccines based on protein assemblies, it is necessary to understand the basic structural features which drive protein self-assembly and functionalization of portions of pathogens. This review highlights recent developments and future perspectives in production of non-viral protein assemblies with essential structural features of subunit vaccines.     

Engineering of Lipid Nanoparticles by the Multifunctionalization of the Surface with Amino Acid Derivatives for the Neutralization of a Target Toxic Peptide

Protein affinity reagents (e.g., antibodies) are often used for basic research, diagnostics, separations, and disease therapy. Although a lot of “synthetic” protein affinity reagents have been developed as a cost-effective alternative to antibodies, their low biocompatibility is a considerable problem for clinical application. Lipid nanoparticles (LNP) represent a highly biocompatible drug delivery agent. However, little has been reported that LNP itself works as a protein affinity reagent in living animals. Here, LNP is engineered for binding to and neutralizing a target toxic peptide in living animals by multifunctionalization with amino acid derivatives. Multifunctionalized LNP (MF-LNP) is prepared using amino acid derivative-conjugated lipids. Optimized MF-LNP exhibits nanomolar affinity to the target toxic peptide and inhibits toxic peptide-dependent hemolysis and cytotoxicity. In addition, MF-LNP captures and neutralizes the toxic peptide after intravenous injection in the bloodstream; in addition, MF-LNP does not release the toxic peptide in the accumulated organ. These results reveal the potential of using LNP as a highly biocompatible protein affinity reagent such as an antidote.     

Identification of enzyme responsible for erythritol utilization and reaction product in yeast Lipomyces starkeyi

We have identified the enzyme responsible for erythritol utilization and its reaction product in the yeast Lipomyces starkeyi CBS 1807. The enzyme, a polyol dehydrogenase requiring NAD+ as a coenzyme, was induced by erythritol in this yeast. We confirmed that the enzyme product was L-erythrulose by MS, NMR, and polarimeter analyses, meaning that we clarified the first step of erythritol utilization in yeasts for the first time. In the case of the oxidative reaction, D-threitol, (2R,3R)-2,3-butanediol, and erythritol were much better substrates than 21 other polyols tested. These three substrates are tetroses and have an R configuration at C-3, and whose third carbon results in easiest oxidation in this enzyme. The research of the substrate specificity in the reductive reaction demonstrated that L-erythrulose and dihydroxyacetone were better substrates, that D-acetoin was inactive and L-erythrose (aldose) was slightly active.     

Detection of pathogenic bacteria by using zinc finger protein fused with firefly luciferase

We constructed a novel bacterial genome detection system using zinc finger protein (ZF) fused with firefly luciferase (ZF-luciferase). Taking advantage of the direct recognition of double-stranded DNA (dsDNA) by ZF, we previously constructed bacterial genome detection systems that did not require dehybridization processes. To detect polymerase chain reaction (PCR) products rapidly and with a high sensitivity, we constructed two kinds of ZF-luciferase, Sp1-fused luciferase (Sp1-luciferase), and Zif268-fused luciferase (Zif268-luciferase). ZF-luciferase not only maintains luciferase activity but also shows dsDNA-binding ability and specificity. Furthermore, we succeeded in the detection of 10 copies of the genome of Legionella pneumophila and Escherichia coli O157. ZF-luciferase would be a useful tool for highly sensitive detection of pathogenic bacterial genome.     

The influence of the organic/inorganic interface on the organic-inorganic hybrid solar cells

Organic-inorganic hybrid solar cells based on poly(3-hexylthiophene) (P3HT) and (6,6)-phenyl C61 butyric acid methyl ester (PCBM) hybridized with ZnO nanorods were fabricated by growing vertical ZnO nanorods on indium tin oxide (ITO) substrates and filling with bulk heterojunction polymers (P3HT:PCBM). The interface between the organic and inorganic nanostructures influences the performance of the organic-inorganic hybrid solar cells. In this paper, the influence of the state of the P3HT:PCBM/ZnO interface on the performance of organic-inorganic hybrid solar cells is examined. The solar cell performance was high when the P3HT:PCBM/ZnO junction area was large. The charge separation is effective when the active layer/electron transport layer junction area is large, resulting in increasing photocurrent and a high conversion efficiency. The bulk-heterojunction polymer concentration was kept low to infiltrate into the ZnO nanorods, resulting in a large active layer/electron transport layer junction area.     

The thermodynamic database malt

The objectives of the development of the thermodynamic database MALT are described together with its main characteristic features. The database, which contains about 5000 species, also composes software for calculation of chemical equilibria and construction of generalized chemical potential diagrams. Discussion is made on the quality of the thermodynamic data to be used in the advanced software for calculating complex chemical equilibria in multicomponent systems and the importance of the chemical potential diagrams for checking the interconsistency of data in a visual manner is stressed.     

Gasification of bean curd refuse with carbon supported noble metal catalysts in supercritical water

Gasification of bean curd refuse was examined in the presence of carbon supported noble metal catalysts in sub- and supercritical water from 473 K to 673 K. The activity of catalyst was in the order of Ru/C >> Rh/C > Pt/C > Pd/C. The effect of reaction condition with Ru/C catalyst was examined in detail. In the presence of catalyst and water, bean curd refuse decomposed to intermediates and then intermediates were gasified. The main gas products were methane and carbon dioxide. The sum of the yield of gases significantly increased with increase in water density at 673 K. The highest total gas yield in a carbon basis was 76% at 673 K and 0.5 g/cm³ of water density. The yield of gases increased with increasing temperature especially above 623 K at initial water density of 0.5 g/cm³. The experiments changing temperature profile revealed that rapid heating was effective for the gasification because rapid heating probably prevented the formation of heavier components in low temperature region.     

Constructing low emitting power systems through grid extension in Papua New Guinea (PNG) with rural electrification

The effective rural electrification method varies with economic status and geographical location, and the benefits of decentralized generation differ for each energy system depending on its characteristics. This paper evaluates the most effective generation strategies with rural electrification in an optimized power system of Papua New Guinea (PNG) using a linear programming model. The energy system model developed for the study includes decentralized generation, centralized generation, and grid systems of electricity and gas with consideration for the current energy system and infrastructure. Two methods of rural electrification, decentralized generation and grid extension, are compared with and without the Clean Development Mechanism (CDM). The results of simulations show that extending the power grid that allows economical generation such as coal-fired power and hydropower to supply rural electricity is a more cost effective way for rural electrification. Although grid extension is more capital intensive than decentralized generation, the former reduces the total system cost through reduction of the fossil fuel use. Extending the power grid is also effective at attracting CDM investments, since it makes the power system flexible and provides opportunities to advance low emitting energy such as hydropower.     

蒙药材止泻木子生物碱的分离与纯化

以夹竹桃科植物止泻木子的干燥种子为原材料,采用乙醇提取法对其生物碱类物质进行粗提取,通过硅胶柱层析和重结晶技术对生物碱粗提物进行分离和纯化。利用核磁共振波谱和质谱法对其一种生物碱单体进行化学结构和分子量鉴定。实验结果表明,用70%乙醇在60℃条件下静止浸取3次,每次提取6 h,得到止泻木子总生物碱提取物,其得率达到0.244%。经过硅胶柱分离和重结晶纯化,可获得3种生物碱单体。对HPLC检测纯度达到94.89%的2号生物碱单体进行核磁共振波谱和质谱检测,得知该生物碱单体是一个分子量为356.59的锥丝碱（conessine）。该结果有利于了解以止泻木子为主要成分的蒙药药剂作用机理。