A novel self-cleaving phasin tag for purification of recombinant proteins based on hydrophobic polyhydroxyalkanoate nanoparticles

A novel protein purification method was developed using microbial polyhydroxyalkanoates (PHA) granule-associated protein phasin, a pH-inducible self-cleaving intein and PHA nanoparticles. Genes for the target proteins to be produced and purified were fused to genes of intein and phasin, the genes were jointly over-expressed in vivo, such as in E. coli cells in this study. The fused proteins containing target protein, intein and phasin produced by the recombinant E. coli were released together with all other E. coli proteins via a bacterial lysis process. They were then adsorbed in vitro to the surfaces of the hydrophobic polymer nanoparticles incubated with the cell lysates. The nanoparticles attached with the fused proteins were concentrated via centrifugation. Then, the reasonably purified target protein was released by self-cleavage of intein and separated with nanoparticles by a simple centrifugation process. Using this system, enhanced green fluorescent protein (EGFP), maltose binding protein (MBP) and beta-galactosidase were successfully purified in their active forms with reasonable yields, respectively, demonstrating the effectiveness and reliability of this purification system. This method allows the production and purification of high value added proteins in a continuous way with low cost.     

Experimental and Theoretical Study of Phosphine-Catalyzed Reaction Modes in the Reaction of α-Substituted Allenes with Aryl Imines

A new phosphine-catalyzed reaction of α-substituted allenes with aryl imines, in stark contrast to classic cycloaddition reactions, has been developed. This reaction delivers valuable highly functionalized itaconimides with excellent stereoselectivities by a new «un-cyclizing» reaction mode involving β′-carbon of α-substituted allenes. Moreover, the present «un-cyclizing» reaction can also be carried out in a one-pot fashion and scaled up to the gram scale by using aryl aldehydes, without the need to isolate the aryl imines. Mechanistic studies and control experiments reveal the crucial role of H₂CO₃ for the present reaction mode. In addition, density functional theory (DFT) calculations were performed to understand the possible mechanism.     

Chiral Iron Porphyrins Catalyze Enantioselective Intramolecular C(sp3)−H Bond Amination Upon Visible-Light Irradiation

Iron-catalyzed asymmetric amination of C(sp³)−H bonds is appealing for synthetic applications due to the biocompatibility and high earth abundance of iron, but examples of such reactions are sparse. Herein we describe chiral iron complexes of meso- and β-substituted-porphyrins that can catalyze asymmetric intramolecular C(sp³)−H amination of aryl and arylsulfonyl azides to afford chiral indolines (29 examples) and benzofused cyclic sulfonamides (17 examples), respectively, with up to 93 % ee (yield: up to 99 %) using 410 nm light under mild conditions. Mechanistic studies, including DFT calculations, for the reactions of arylsulfonyl azides reveal that the Fe(NSO₂Ar) intermediate generated in situ under photochemical conditions reacts with the C(sp³)−H bond through a stepwise hydrogen atom transfer/radical rebound mechanism, with enantioselectivity arising from cooperative noncovalent interactions between the Fe(NSO₂Ar) unit and the peripheral substituents of the chiral porphyrin scaffold.     

Solar PAR and UV radiation affects the physiology and morphology of the cyanobacterium Anabaena sp. PCC 7120

Solar UV radiation (280-400 nm) may affect morphology of cyanobacteria, however, little has been evidenced on this aspect while their physiological responses were examined. We investigated the impacts of solar PAR and UVR on the growth, photosynthetic performance and morphology of the cyanobacterium Anabaena sp. PCC7120 while it was grown under three different solar radiation treatments: exposures to (a) constant low PAR (photosynthetic active radiation, 400-700 nm), (b) natural levels of solar radiation with and (c) without UV radiation (290-400 nm). When the cells were exposed to solar PAR or PAR+UVR, the photochemical efficiency was reduced by about 40% and 90%, respectively, on day one and recovered faster under the treatment without UVR over the following days. Solar UVR inhibited the growth up to 40%, reduced trichome length by up to 49% and depressed the differentiation of heterocysts. Negligible concentrations of UV-absorbing compounds were found even in the presence of UVR. During the first 2 d of exposure to natural levels of PAR, carotenoid concentrations increased but no prolonged increase was evident. Heterocyst formation was enhanced under elevated PAR levels that stimulated quantum yield and growth after an initial inhibition. Higher concentrations of carotenoids and a twofold increase in the carotenoid to chlorophyll a ratio provided protection from the high levels of solar PAR. Under radiation treatments with UVR the relatively greater decrease in chlorophyll a concentrations compared with the increase in carotenoids was responsible for the higher carotenoid: chlorophyll a ratio. Heterocyst formation was disrupted in the presence of solar UVR. However, the longer term impact of heterocyst disruption to the survival of Anabaena sp. requires further study.     

Low-intensity ultrasound enhances the antimicrobial activity of neutral peptide TGH2 against Escherichia coli

In recent years, foodborne diseases caused by Escherichia coli are a major threat to the food industry and consumers. Antimicrobial peptides (AMPs) and ultrasound both have good inhibitory effects on E. coli. In this work, the mechanism of action and synergistic effect of an in silico predicted AMP, designated as TGH2 (AEFLREKLGDKCTDRHV), from the C-terminal sequence of Tegillarca granosa hemoglobin, combined with low-intensity ultrasound was explored. The minimal inhibitory concentration (MIC) of TGH2 on E. coli decreased by 4-fold to 31.25 μg/mL under 0.3 W/cm² ultrasound treatment, while the time kill curve analysis showed that low-intensity ultrasound combined with peptide TGH2 had an enhanced synergistic bactericidal effect after 0.5 h. The permeability on E. coli cell membrane increased progressively during combined treatment with peptide TGH2 and low-intensity ultrasound, resulting in the leakage of intracellular solutes, as shown by transmission electron microscopy (TEM). Structural analysis using circular dichroism (CD) revealed that peptide TGH2 has an α-helical structure, showing a slight untwisting effect under 0.3 W/cm² ultrasound treatment for 0.5 h. The findings here provide new insight into the potential application of ultrasound and AMPs combination in food preservation.     

A pair of spaces of upper semi-continuous maps and continuous maps

For a Tychonoff space X, we use ↓USC(X) and ↓C(X) to denote the families of the regions below all upper semi-continuous maps and of the regions below all continuous maps from X to I=[0,1], respectively. In this paper, we consider the spaces ↓USC(X) and ↓C(X) topologized as subspaces of the hyperspace Cld(X×I) consisting of all non-empty closed sets in X×I endowed with the Vietoris topology. We shall prove that ↓USC(X) is homeomorphic (≈) to the Hilbert cube Q=ω[−1,1] if and only if X is an infinite compact metric space. And we shall prove that (↓USC(X),↓C(X))≈(Q,c₀), where , if and only if ↓C(X)≈c₀ if and only if X is a compact metric space and the set of isolated points is not dense in X.     

Metabolic engineering for microbial production and applications of copolyesters consisting of 3-hydroxybutyrate and medium-chain-length 3-hydroxyalkanoates

Poly(hydroxyalkanoate)s (PHAs) are a class of microbially synthesized polyesters that combine biological properties, such as biocompatibility and biodegradability, and non-bioproperties such as thermoprocessability, piezoelectricity, and nonlinear optical activity. PHA monomer structures and their contents strongly affect the PHA properties. Using metabolic engineering approaches, PHA structures and contents can be manipulated to achieve controllable monomer and PHA cellular contents. This paper focuses on metabolic engineering methods to produce PHA consisting of 3-hydroxybutyrate (3HB) and medium-chain-length 3-hydroxyalkanoates (3HA) in recombinant microbial systems. This type of copolyester has mechanical and thermal properties similar to conventional plastics such as poly(propylene) and poly(ethylene terephthalate) (PET). In addition, pathways containing engineered PHA synthases have proven to be useful for enhanced PHA production with adjustable PHA monomers and contents. The applications of PHA as implant biomaterials are briefly discussed here. In the very near term, metabolic engineering will help solve many problems in promoting PHA as a new type of plastic material for many applications.     

一种含噻吩环酰腙及其Cu(Ⅱ)配合物的合成、晶体结构和抗氧化活性及量化计算

合成了一种酰腙类含硫Schiff碱4-(二乙胺基)水杨醛缩2-噻吩甲酰腙一水合物(H₂L·H₂O,1),采用扩散法制备了它的铜配合物[Cu(phen)L](2,phen=菲咯啉),通过元素分析、红外光谱、紫外光谱和X射线单晶衍射对它们进行了表征。1的晶体属正交晶系,P2₁2₁2空间群;2的晶体属单斜晶系,P2₁/c空间群。在2中H₂L以烯醇式脱除羟基质子与铜配位,中心离子的配位环境为畸变的四方锥构型。应用Gaussian09程序,采用密度泛函方法(DFT),分别在B3LYP/6-31G(d)和UB3LYP/6-31G(d)水平对1和2进行了量化计算,对分子的前线分子轨道的能量和组成以及电子光谱进行了分析讨论。测试、分析了化合物的抗氧化活性。     

Boron Nitride Nanomaterials for Thermal Management Applications

Hexagonal boron nitride nanosheets (BNNs) are analogous to their two‐dimensional carbon counterparts in many materials properties, in particular, ultrahigh thermal conductivity, but also offer some unique attributes, including being electrically insulating, high thermal stability, chemical and oxidation resistance, low color, and high mechanical strength. Significant recent advances in the production of BNNs, understanding of their properties, and the development of polymeric nanocomposites with BNNs for thermally conductive yet electrically insulating materials and systems are highlighted herein. Major opportunities and challenges for further studies in this rapidly advancing field are also discussed.     

Minimal sets in compact connected subspaces

Let X be a topological space, f:X→X be a continuous map, and Y be a compact, connected and closed subset of X. In this paper we show that, if the boundary X_∂Y contains exactly one point v and f(v)∈Y, then Y contains a minimal set of f.