Exploration of GH94 Sequence Space for Enzyme Discovery Reveals a Novel Glucosylgalactose Phosphorylase Specificity

The substantial increase in DNA sequencing efforts has led to a rapid expansion of available sequences in glycoside hydrolase families. The ever-increasing sequence space presents considerable opportunities for the search for enzymes with novel functionalities. In this work, the sequence-function space of glycoside hydrolase family 94 (GH94) was explored in detail, using a combined approach of phylogenetic analysis and sequence similarity networks. The identification and experimental screening of unknown clusters led to the discovery of an enzyme from the soil bacterium Paenibacillus polymyxa that acts as a 4-O-β-d -glucosyl-d -galactose phosphorylase (GGalP), a specificity that has not been reported to date. Detailed characterization of GGalP revealed that its kinetic parameters were consistent with those of other known phosphorylases. Furthermore, the enzyme could be used for production of the rare disaccharides 4-O-β-d -glucosyl-d -galactose and 4-O-β-d -glucosyl-l -arabinose. Our current work highlights the power of rational sequence space exploration in the search for novel enzyme specificities, as well as the potential of phosphorylases for rare disaccharide synthesis.     

激光在线测厚振动分析与精度优化

激光测厚具有安全可靠、测量精度高、测量范围大等优点,广泛应用于纸张、电池极片等薄膜类材料厚度的在线测量。带材宽幅方向扫描测厚时由于扫描架往复运动会产生机械振动,影响在线测厚精度。针对该问题,以锂离子电池极片厚度测量为例,使用双激光差动式测厚平台对电池极片和铜箔分别进行厚度测量,然后对测厚数据进行频谱分析,探究其振动规律的相似性,并基于频谱分析结果采用滑动带阻滤波方式对测厚数据进行处理,滤波后极片和铜箔的厚度极差分别降低了33.4%和73.8%,有效过滤了机械振动导致的测量误差,可满足极片和铜箔厚度测量的精度要求。     

Rational Design of Potent Inhibitors of a Metallohydrolase Using a Fragment-Based Approach

Metallohydrolases form a large group of enzymes that have fundamental importance in a broad range of biological functions. Among them, the purple acid phosphatases (PAPs) have gained attention due to their crucial role in the acquisition and use of phosphate by plants and also as a promising target for novel treatments of bone-related disorders and cancer. To date, no crystal structure of a mammalian PAP with drug-like molecules bound near the active site is available. Herein, we used a fragment-based design approach using structures of a mammalian PAP in complex with the Maybridge^TM fragment CC063346, the amino acid L-glutamine and the buffer molecule HEPES, as well as various solvent molecules to guide the design of highly potent and efficient mammalian PAP inhibitors. These inhibitors have improved aqueous solubility when compared to the clinically most promising PAP inhibitors available to date. Furthermore, drug-like fragments bound in newly discovered binding sites mapped out additional scaffolds for further inhibitor discovery, as well as scaffolds for the design of inhibitors with novel modes of action.     

Effects of Bacillus velezensis zk1 on the physiology and metabolism of peaches

Bacillus velezensis zk1 is the dominant bacterium that causing rot in peaches. However, the mechanisms through that this bacterium causes rot have not been elucidated. Here, we explored the mechanisms of peach decay caused by B. velezensis zk1. The invasion of B. velezensis zk1 in peaches resulted in an increase in glucose and arabinose contents in fruit tissues. Moreover, the relative conductivity of the fruit reached 84% after 4 days of culture with bacterial invasion. With the destruction of cells, the malondialdehyde content increased, whereas the vitamin C, dialdehyde, flavonoid and total phenol contents decreased. Polyphenol oxidase, superoxide dismutase, catalase, peroxidase, ascorbate peroxidase and ammonia lyase activities also decreased. Overall, these findings demonstrated that B. velezensis zk1 infection damaged peach chloroplasts, mitochondria, respiratory chain activity and related free radical scavenging enzyme systems, thereby disrupting the normal physiological metabolism of peaches and causing rot.     

微波辅助孵育法富集麦胚多肽的工艺优化

目的 麦胚孵育过程中蛋白酶将蛋白质水解成肽和氨基酸，多肽具有明确的生理活性和营养调节作用。本研究以麦胚为试验原料，采用微波辅助预处理的方法，研究孵育的温度、时间、pH及料液比4种因素对孵育后的蛋白酶活力及多肽含量的影响。 方法 通过单因素和响应曲面试验进行工艺条件优化。 结果 与未进行微波辅助预处理的样品相比，微波辅助处理（600 W, 10 s）能显著提高孵育后样品中的蛋白酶酶活力（约9.4倍）和肽含量（约3.1倍）。经过微波辅助处理后，孵育温度为51.5℃、pH为4.0、时间为6.33 h、料液比为1:7时，蛋白酶活力达最高为3826.24 U/g；孵育温度为45.0 ℃、pH为4.8、时间为8 h、料液比为1:7时，肽含量达最高为262.63 mg/g。 结论 微波辅助处理能有效的激活麦胚孵育液中的内源性蛋白酶活性，促进蛋白水解反应，显著提高孵育后的肽含量。该研究结果为麦胚多肽的制备新工艺开发提供了研究基础。     

Identification and U-control of a state-space system with time-delay

This article presents a state-space model with time-delay to map the relationship between known input-output data for discrete systems. For the given input-output data, a model identification algorithm combining parameter estimation and state estimation is proposed in line with the causality constraints. Consequently, this article proposes a least squares parameter estimation algorithm, and analyzes its convergence for the studied systems to prove that the parameter estimation errors converge to zero under the persistent excitation conditions. In control system design, the U-model based control is introduced to provide a unilateral platform to improve the design efficiency and generality. A simulation portfolio from modeling to control is provided with computational experiments to validate the derived results.     

基于光谱尺度空间与管道滤波的红外目标检测

为了准确地从复杂干扰背景下检测出真实弱小目标，本文引入视觉显著性，设计了基于快速光谱尺度空间与动态管道滤波的红外目标检测算法。基于真实目标与背景内容之间的整体差异，引入快速光谱尺度空间与阈值分割技术，设计视觉显著性机制，对红外图像完成处理，输出全局显著性映射，以高效过滤干扰背景内容。考虑目标与背景的局部特征差异，构建自适应局部对比度增强机制，对粗检测结果实施处理，获取对应的局部显著性映射，改善视觉显著性区域内目标的对比度。引入高斯差分理论，通过估算每一帧红外图像中的目标像素直径，形成动态管道滤波，充分消除虚警，准确识别出弱小目标。多组实验数据显示：较已有的红外目标检测技术而言，在各种不同的复杂背景干扰下，所提算法呈现出更好的检测能力，拥有更理想的接收机工作特性ROC曲线。     

Synergistic effect of high-intensity ultrasound and β-cyclodextrin treatments on browning control in apple juice

This work evaluated the synergistic effects of combined high-intensity ultrasound (HIU) with β-cyclodextrin (β-CD) treatments on inhibiting browning of apple juice and explored the mechanism through simulation system. The combined treatment of 300 W HIU with 0.006 g mL⁻¹ β-CD had a synergistic impact on maintaining juice colour, resulting in a 39.06% reduction in browning degree, only a 36.64% decrease in total phenolic content, and a 17.82% reduction in PPO activity. The inhibition of enzymatic browning in simulated system revealed that HIU suppressed the enzyme (Polyphenol oxidase, PPO) and β-CD inhibited enzyme (PPO) and embedded substrate (polyphenol). The results of spectroscopic analysis showed that the particle-size distribution of PPO narrowed, the content of α-helix in the secondary structure increased, the fluorescence intensity increased, and the maximum wavelength was red-shifted after HIU and β-CD treatment. Changes in structure could further result in PPO activity loss. Hence, the combined treatment could synthetically alleviate the browning of apple juice.