A High-Throughput Screening System Based on Fluorescence-Activated Cell Sorting for the Directed Evolution of Chitinase A

Chitinases catalyze the degradation of chitin, a polymer of N-acetylglucosamine found in crustacean shells, insect cuticles, and fungal cell walls. There is great interest in the development of improved chitinases to address the environmental burden of chitin waste from the food processing industry as well as the potential medical, agricultural, and industrial uses of partially deacetylated chitin (chitosan) and its products (chito-oligosaccharides). The depolymerization of chitin can be achieved using chemical and physical treatments, but an enzymatic process would be more environmentally friendly and more sustainable. However, chitinases are slow-acting enzymes, limiting their biotechnological exploitation, although this can be overcome by molecular evolution approaches to enhance the features required for specific applications. The two main goals of this study were the development of a high-throughput screening system for chitinase activity (which could be extrapolated to other hydrolytic enzymes), and the deployment of this new method to select improved chitinase variants. We therefore cloned and expressed the Bacillus licheniformis DSM8785 chitinase A (chiA) gene in Escherichia coli BL21 (DE3) cells and generated a mutant library by error-prone PCR. We then developed a screening method based on fluorescence-activated cell sorting (FACS) using the model substrate 4-methylumbelliferyl β-d-N,N′,N″-triacetyl chitotrioside to identify improved enzymes. We prevented cross-talk between emulsion compartments caused by the hydrophobicity of 4-methylumbelliferone, the fluorescent product of the enzymatic reaction, by incorporating cyclodextrins into the aqueous phases. We also addressed the toxicity of long-term chiA expression in E. coli by limiting the reaction time. We identified 12 mutants containing 2–8 mutations per gene resulting in up to twofold higher activity than wild-type ChiA.     

纳米级超声造影剂的理论研究进展

为了克服超声造影剂中微米级气泡尺寸较大的局限性,大量研究人员对超声应用的替代造影剂(纳米级造影剂)进行了研究。随着生物纳米技术的飞速发展,纳米级超声造影剂在诊断与治疗领域有着广阔的发展前景。与超声造影剂中的微米级气泡相比,纳米级造影剂粒径较小,渗透能力极强,可以通过血管内皮间隙,进而可以实现血管外病变部位的显影。文中详细论述了超声造影剂在超声作用下的行为以及2种主要的纳米级造影剂:纳米气泡和纳米液滴造影剂,对其理论研究进展进行了总结,并提出了目前仍存在的一些问题及其未来的研究方向。     

Hydrodynamics study of a fast liquid–liquid oxidation process with in situ gas production in microreactors

Hydrodynamics characteristics of a fast and highly exothermic liquid–liquid oxidation process with in situ gas production in microreactors were studied using a newly developed experimental method. In the adipic acid synthesis through the K/A oil (the mixture of cyclohexanol and cyclohexanone) oxidation with nitric acid, bubble generation modes were divided into four categories. The gas production became more intensive, unstable, even explosive with increasing the oil phase feed rate and the temperature. A novel automatic image processing method was developed to monitor the instantaneous velocity online by tracking the gas–liquid interface. The axial velocity at the same location was unstable due to the changing gas production rate. Furthermore, the actual residence time was obtained easily with being only 36% of the space–time minimally, beneficial for establishing accurate kinetics and mass transfer models with time participation. Finally, an empirical correlation was developed to predict the actual residence time under different conditions.     

Taylor Bubble Study of the Influence of Fluid Dynamics on Yield and Selectivity in Fast Gas-Liquid Reactions

A consecutive competitive gas-liquid reaction is investigated using a Taylor bubble setup regarding the influence of fluid mixing in the bubble wake on yield and selectivity. The concentration fields behind a Taylor bubble are visualized and measured quantitatively with a novel time-resolved absorption imaging technique based on Beer Lamberts law and an integral selectivity is derived. In addition, the calculation of the local selectivity, often used in numerical approaches, is discussed and the existing experimental limits for its derivation are pointed out. Finally, an increase in selectivity of a competitive consecutive reaction for enhanced mixing is experimentally confirmed.     

Formation process of zirconia nanotubes and porous structures and model of oxygen bubble growth

Preparation and growth mechanism of anodization of Ti and Al has been widely concerned for two decades, but the research on anodic ZrO₂ is relatively lacking. In this paper, anodic TiO₂ and ZrO₂ nanotubes were prepared in glycerol electrolyte containing 0.35 M NH₄F and 4 vol% H₂O under different anodizing voltages. We had successfully prepared the anodic ZrO₂ nanotubes (AZNTs) with a complete top and a “bulb” at the bottom under 60 V, and with the increase of the applied anodizing voltage, the “bulb” cavity also increased. However, under the same anodizing conditions, the surface of anodic TiO₂ nanotubes (ATNTs) is a cluster of nano-tip morphology, and the bottom of the ATNTs is a conventional hemisphere shape. In addition, both AZNTs and porous anodic zirconia (PAZ) were found to coexist in the anodic ZrO₂ layer prepared at 60 V. Here, we used the oxygen bubble model and ionic current and electronic current theories to analyze the reason of the special morphology. It is confirmed that the porous anodic oxides are actually evolved from nanotubes. In other words, the structure is essentially the same.     

建筑结构控制爆破拆除的力学分析与应用

应用运动学、动力学理论建立建(构)筑物控爆坍落过程的力学模型,进而研究在非定常约束条件下坍落结构的力学特征。首次提出了采用延时起爆技术时,非自由质点系及其广义坐标应是建立在每一段位上的瞬间虚位移。在此基础上,分析求解运动过程中时控单元的质点自由度,为精确选定爆破部位及时序提供了设计理论和计算依据。同时,结合振动理论,将控爆时差、结构载荷分布调制为振动系统,形成坍落结构梁板单元的强迫振动,充分实现在运动中进一步解体、破碎其自身结构,并形成具有明显缓冲效果的下落过程。应用实践结果证明力学分析是正确的,为工程结构爆破拆除计算机辅助设计及动态模拟技术的有效应用奠定了基础。     

A novel pre-dilution,swirling jet diffuser to enhance effluent mixing: Hydrodynamics and dilution performance

Diffusers are widely-used to quickly dilute effluents in receiving water bodies. This study proposed a novel diffuser that pre-mixes effluent with ambient water before discharging and that uses the swirling jet to further enhance near-field dilution. The nozzle of the diffuser was examined in two ambient flow conditions: co-flow and counter-flow that are commonly-met in the environment such as oceans due to tidal effect. Physical experiments were first conducted in co-flow on its dilution performance and hydrodynamics, using heated water as the effluent. A 3-D CFD model was developed and calibrated the co-flow scenarios, and then used to investigate the diffuser in counter-flow. The results showed that the nozzle can effectively reduce the maximum temperature rise of the effluent by about 50 % before discharging. The swirling jet from the outlet has a larger shear area, half-width and entrainment rate, enabling the effluent to be rapidly diluted to a minimum of around 10 times at x/D = 6 in co-flow, whereas the dilution for conventional nozzles is about 1 because of the potential core. The flow amplification ratio (α) decreases gradually with increasing velocity ratio in co-flow but increases with increasing velocity ratio in counter-flow. The counter-flow reduces the water drawn into the device; however, the pre-dilution effect at the outlet remains stable. The near-field dilution in counter-flow was significantly enhanced than that in co-flow. Environmental regulations at outfalls and mixing zones can be more easily met using this novel diffuser.     

以新发展理念引领高校专业基础课教学改革

以“创新、协调、绿色、开放、共享”为内核的新发展理念，是对马克思主义发展理念的继承和发扬，极具时代精神，富含问题意识，为高校思想政治教育发展、教育教学改革实践提供了强大的理论支撑。本文以“通信原理”为例，阐述了新发展理念在课程改革中的思路和方法，实现了思想政治教育与专业基础课程有机融合，为深化高校教学改革、创新人才培养模式提供了思路。     

Simulation and exploration of cavitation process during microalgae oil extracting with ultrasonic-assisted for hydrogen production

Microalgae is promising to be used as feedstock resources for hydrogen production due to its high oil and grease contents. This promotes the development of extraction technology of microalgae oil. In this study, based on the Rayleigh-Plesset equation, the effects of temperature, pressure, ultrasonic power and frequency on the bubble motion of ethanol ultrasound cavitation are investigated. Subsequently, the effects of different process parameters on the extraction rate are studied using Schizochytrium sp. as raw material by stirring or ultrasonic-assisted extraction. And the composition of algae extraction oil is analyzed. The results show that the amplitude of cavitation bubbles increases with the increase of ultrasonic power and decrease of ultrasonic frequency. The extraction rate of algae oil reaches 93.76 ± 0.48% when the ultrasonic power is 150 W, the reaction time is 30 min, the temperature is 50 °C and the liquid-solid ratio is 10:1.     

Controlled galvanic decoration boosting catalysis: Enhanced glycerol electro-oxidation at Cu/Ni modified macroporous films

We report on glycerol electro-oxidation in alkaline medium at macroporous Ni electrodes decorated with Cu particles. Macroporous Ni film is electrodeposited, using hydrogen bubbles as dynamic templates, atop of a Cu substrate. This film shows good electrocatalytic activity towards glycerol oxidation reaction (GOR). The Ni film is further decorated with Cu via spontaneous deposition from CuSO₄ solution. This is done to enhance the catalytic activity of the film towards GOR. The morphology of the Cu-decorated Ni film is controlled using various additives such as KCl and (NH₄)₂SO₄ which are added to the Cu deposition bath. The as-prepared Cu-decorated Ni films are characterized by electrochemical measurements, scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS). It is found that these additives have tremendous effects on the morphology and the electrocatalytic activity of the decorating Cu particles.The decorated Ni foam showed superior electrocatalytic activity towards the GOR, as confirmed by the negative shift in the onset oxidation potential (ca. 100 mV) together with an increase in oxidation current that is up to 1.5-fold during the cyclic voltammetry (CV) measurements, compared to the undecorated Ni foam.