Covalent Linkage of an R-ω-Transaminase to a d-Amino Acid Oxidase through Protein Splicing to Enhance Enzymatic Catalysis of Transamination

R-ω-Transaminases (RTAs) catalyse the conversion of R-configured amines [e.g., (R)-1-phenylethylamine] into the corresponding ketones (e.g., acetophenone), by transferring an amino group from an amino donor [e.g., (R)-1-phenylethylamine] onto an amino acceptor (e.g., pyruvate), resulting in a co-product (e.g., d -alanine). d -Alanine can be deaminated back to pyruvate by d -amino acid oxidase (DAAOs). Here, through in vivo subunit splicing, the N terminus of an RTA subunit (RTA^S) was specifically ligated to the C terminus of a DAAO subunit (DAAO^S) through native peptide bonds (RTA&DAAO). RTA^S is in close proximity to DAAO^S, at a molecular-scale distance. Thus the transfer of pyruvate and d -alanine between RTA and DAAO can be directional and efficient. Pyruvate→d -alanine→pyruvate cycles are efficiently formed, thus promoting the forward transamination reaction. In a different, in vitro noncovalent approach, based on coiled-coil association, the RTA^S N terminus was specifically associated with the DAAO^S C terminus (RTA#DAAO). In addition, the two mixed individual enzymes (RTA+DAAO) were also studied. RTA&DAAO has a shorter distance between the paired subunits (RTA^S–DAAO^S) than RTA#DAAO, and the number of the paired subunits is higher than in the case of RTA#DAAO, whereas RTA+DAAO cannot form the paired subunits. RTA&DAAO exhibited a transamination catalysis efficiency higher than that of RTA#DAAO and much higher than that of RTA+DAAO.     

Effect of Operating Parameters on Gas‐Solid Hydrodynamics and Heat Transfer in a Spouted Bed

Through a combined computational fluid dynamics and discrete element method approach, the effect of the operating parameters on the hydrodynamics and heat‐transfer properties of gas‐solid two‐phase flows in a spouted bed are extensively investigated. Considering the high velocity in the fountain region, gas turbulence is resolved by employing the large‐eddy simulation. The rolling friction model is adopted for more precise predictions of solid behavior near the wall. Subsequently, the gas‐solid flow patterns, gas‐solid velocities, and temperature evolution are investigated. Moreover, different operating conditions and geometry configurations are evaluated with respect to heat‐transfer performance. The results provide a fundamental understanding of heat‐transfer mechanisms in spouted beds.     

Do IT freelancers increase their entrepreneurial behavior and performance by using IT self-efficacy and social capital? Evidence from Bangladesh

This study applies entrepreneurship theory and social capital perspective to IT freelancing to facilitate a better understanding of it. Our model hypothesizes that both IT self-efficacy and social capital obtained through social media interactions contribute to IT freelancers’ entrepreneurial behavior and then this entrepreneurial behavior influences their performance. We collected primary data from one of the leading online freelancing marketplaces, oDesk.com (currently Upwork.com). Our empirical findings confirm the core assertion of our research model. The implications of these findings for research and practice are discussed. Directions for future research are also provided.     

放射性气载流出物取样代表性分析

为获取核设施放射性气载流出物单点连续监测取样位置,提出基于随机轨道模型(DRW)的气-固多相湍流耦合方法求解代表性取样区域。借助k-epsilon湍流模型模拟连续相,并引入离散颗粒模型(DPM)模拟离散相,建立基于DRW模型的排风管道内流道气-固多相湍流耦合计算模型,计算了核设施气载流出物在管道内流道流场分布规律,分析了内流道流体气旋角、气流速度、示踪气体浓度、气溶胶粒子浓度与管道高度间的关联关系。分析结果表明,随着截面高度的增加,气旋角、气流速度变异系数(COV)、示踪气体浓度COV及示踪气体浓度最大值与平均值的偏差逐渐降低并趋于稳定,气溶胶粒子浓度COV在截面6与截面8满足取样代表性要求;基于计算流体动力学方法可快速地确定出代表性取样位置,为气载流出物单点取样现场试验提供了理论参考依据。     

基于超材料的无标记光学生物传感

超材料(metamaterials)因为能够在亚波长尺度范围内精细调控电磁波而受到人们广泛关注。超材料具有丰富的电磁模态,在表面支持高度局域场增强且对周围介电环境极其敏感,可应用于无标记光学生物传感领域。与传统光学生物传感器相比,超材料生物传感器具有小型化、集成化、高度灵敏、多功能可定制等突出优点。本文总结了近年来超材料生物传感器在可见光、近红外、中红外以及太赫兹波段的研究进展,包括折射率生物传感、表面增强拉曼散射、表面增强红外吸收和太赫兹生物传感等。     

小型无线场强探测传感器研究

近年来,微波加热因其高效性和清洁无污染等优点广泛应用于各个领域。然而,微波加热的不均匀性限制了微波作为高效加热能源的应用。通过测量和分析加热腔中的电场分布情况可以帮助设计人员改进微波加热腔体设计,提高微波加热的均匀性。现有的场强测量设备均为有线设备,应用场景极为有限。因此,本文提出了一种由探头、接收机和上位机三部分组成的无线场强探测传感器。介绍了无线场强探测传感器的结构和原理,采用横电磁波小室进行校准。通过一系列测量实验表明实测值与标准场强仪测量值一致性较好,可满足工程测量需求。     

凌庄子水厂保水堰流量系数试验

凌庄子水厂蓄水池进水口处有一保水堰,为非标准薄壁堰,不能使用已有堰流公式对其过流量进行准确计算。为了得出较为精确的过流流量,按照重力相似准则制作几何比尺为1∶5的模型进行试验研究。在已有自由出流公式的基础上,对自由出流流量系数进行修正并对淹没情况下流量变化过程进行研究。对该非标准堰自由出流流量系数的实测值与经验值进行分析比较,发现堰板槽降低了实际自由出流过流能力。淹没出流的流量系数主要与下游尾水位有关,试验中形成的淹没式堰流受实际堰型尺寸影响,下游尾水位和堰上水位近似相等,不完全适用已有淹没出流流量公式,通过试验给出了修正淹没系数随h/p的变化关系。结果表明利用堰前、堰后水位初步计算过流流量是可行的,可为该工程提供参考,也可为实际工程中非标准矩形堰的流量计算提供思路。     

水产品及其制品中大西洋鲑鱼实时荧光PCR检测方法建立

目的:三文鱼营养丰富,含有ω-3不饱和脂肪酸,深受大众喜爱,但存在产品标签标示错误、掺伪掺假、以次充好的现象。目前采用检测标准SN/T 3589.7—2013《出口食品中常见鱼类及其制品的鉴伪方法第七部》对鲑鱼成分检测,无法区分大西洋鲑鱼和其他种类的三文鱼。因此有必要建立一种用于区分上述几种三文鱼的快速检测方法。方法:研究建立了针对大西洋鲑鱼成分鉴定的实时荧光PCR法,并对该方法的特异性和检出限进行了验证。结果:该方法具有良好的特异性,检出限为1%(w/w)。结论:可应用于大西洋鲑鱼的日常检测,为监管部门提供技术支持。