Analysis of the Structure and Biosynthesis of the Lipopolysaccharide Core Oligosaccharide of Pseudomonas syringae pv. tomato DC3000

Lipopolysaccharide (LPS), the major component of the outer membrane of Gram-negative bacteria, is important for bacterial viability in general and host–pathogen interactions in particular. Negative charges at its core oligosaccharide (core-OS) contribute to membrane integrity through bridging interactions with divalent cations. The molecular structure and synthesis of the core-OS have been resolved in various bacteria including the mammalian pathogen Pseudomonas aeruginosa. A few core-OS structures of plant-associated Pseudomonas strains have been solved to date, but the genetic components of the underlying biosynthesis remained unclear. We conducted a comparative genome analysis of the core-OS gene cluster in Pseudomonas syringae pv. tomato (Pst) DC3000, a widely used model pathogen in plant–microbe interactions, within the P. syringae species complex and to other plant-associated Pseudomonas strains. Our results suggest a genetic and structural conservation of the inner core-OS but variation in outer core-OS composition within the P. syringae species complex. Structural analysis of the core-OS of Pst DC3000 shows an uncommonly high phosphorylation and presence of an O-acetylated sugar. Finally, we combined the results of our genomic survey with available structure information to estimate the core-OS composition of other Pseudomonas species.     

牛蒡寡糖对黄瓜幼苗诱导抗病性的研究

利用浓度为1．0mg／ml、3．0mg／ml、5．0mg／ml、8．0mg／ml和10．0mg／ml的牛蒡寡糖对黄瓜幼苗离体诱导3d，并用5．0mg／ml的牛蒡寡糖对黄瓜幼苗离体诱导1～6d后，测定与抗病相关的防御酶活性；利用5．0mg／ml的牛蒡寡糖诱导黄瓜幼苗第1真叶3d后接种白粉病菌，调查黄瓜植株的病情指数。结果表明，与对照组相比，不同浓度的牛蒡寡糖对黄瓜幼苗均有诱导作用，能显著提高被处理叶片PAL、POD、PPO、SOD、CAT和壳多糖酶的活性，不同的酶活性达到最高水平时的诱导浓度略有差异。几种酶活性随诱导的时间进程呈现不同水平的动态变化趋势，其中POD活性的提高幅度最大，且一直呈上升趋势。第1真叶的处理组较对照组病情指数降低11．91％，同株未诱导的第2真叶处理组较对照组降低27．42％，说明牛蒡寡糖诱导了黄瓜幼苗对白粉病的系统抗性。     

The Peculiar Structure of Acetobacter pasteurianus CIP103108 LPS Core Oligosaccharide

Acetobacter pasteurianus, a member of the Alphaproteobacteria, is an acetic acid-producing bacterium present on sugar-rich substrates such as such as fruits, flowers and vegetables and traditionally used in the production of fermented food. The preferred living habitat associated with acid conditions makes the structure of the bacterial cell wall interesting to study, due to expected uncommon features. We have used a combination of chemical, analytical and NMR spectroscopy approaches to define the complete structure of the core oligosaccharide from A. pasteurianus CIP103108 LPS. Interestingly, the core oligosaccharide displays a high concentration of negatively charged groups, structural features that might contribute to reinforcing the bacterial membrane.     

Redirecting Killer T Cells through Incorporation of Azido Sugars for Tethering Ligands

The genetic expression of chimeric antigen receptors (CARs) on the surfaces of T cells enables the redirection of T cell specificity. To enhance the versatility of T cells as tumor-specific killers, we developed a nongenetic approach by which azide-containing sialic acids were metabolically incorporated into T cells to modify cellular sialyl glycans. After successful display of these moieties on the T cells, small-molecule ligands such as RGD and folate (as proof-of-concept, rather than supersized antibodies) were clicked orthogonally, leading to highly selective time- and dose-dependent cytotoxicity to integrin α_vβ₃- and folate-receptor-positive cells, respectively. This chemical approach provides a facile platform for rational design of tumor-specific cytotoxic T cells for targeted immunotherapy.     

微波-过氧化氢协同降黏辅助酶法制备壳寡糖

利用微波-过氧化氢协同处理壳聚糖,通过降低壳聚糖的黏度辅助促进其酶解制备壳寡糖。通过单因素实验与正交实验,确定了影响壳聚糖微波-过氧化氢降黏效果的因素主次顺序是微波时间、H2O2的用量、微波功率。优化后的降黏条件是:H2O2添加量为0.1 mL,微波功率为120 W,微波时间为0.5 min,所得壳聚糖黏度比原料黏度降低了96.9%,且并未发生结构的改变。对其酶解的结果表明,经微波-过氧化氢协同降黏再酶解的壳寡糖得率为80.1%,比未经降黏处理的提高了11.8%,产物壳寡糖数均相对分子质量为2 027,平均聚合度为10,明显低于未经降黏处理而直接酶解所得的壳寡糖。     

Molecular Design and Synthesis of a Novel Substrate for Assaying Lysozyme Activity

A novel substrate {Galβ1,4GlcNAcβ1,4GlcNAc-β-pNP [Gal(GlcNAc)₂-β-pNP]} for assaying lysozyme activity has been designed using docking simulations and enzymatic synthesis via β-1,4-galactosyltransferase-mediated transglycosylation from UDP-Gal as the donor to (GlcNAc)₂-β-pNP as the acceptor. Hydrolysis of the synthesized Gal(GlcNAc)₂-β-pNP and related compounds using hen egg-white lysozyme (HEWL) demonstrated that the substrate was specifically cleaved to Gal(GlcNAc)₂ and p-nitrophenol (pNP). A combination of kinetic studies and docking simulation was further conducted to elucidate the mode of substrate binding. The results demonstrate that Gal(GlcNAc)₂-β-pNP selectively binds to a subsite of lysozyme to liberate the Gal(GlcNAc)₂ and pNP products. The work therefore describes a new colorimetric method for quantifying lysozyme on the basis of the determination of pNP liberated from the substrate.     

Glycans in Medicinal Chemistry: An Underexploited Resource

The biological relevance of glycans as mediators of key physiological processes, including disease‐related mechanisms, makes them attractive targets for a wide range of medical applications. Despite their important biological roles, especially as molecular recognition elements, carbohydrates have not been fully exploited as therapeutics mainly due to the scarcity of structure–activity correlations and their non‐drug‐like properties. A more detailed understanding of the complex carbohydrate structures and their associated functions should contribute to the development of new glycan‐based pharmaceuticals. Recent significant progress in oligosaccharide synthesis and chemical glycobiology has renewed the interest of the medicinal chemistry community in carbohydrates. This promises to increase our possibilities to harness them in drug discovery efforts for the development of new and more effective, synthetic glycan‐based therapeutics and vaccines.     

壳寡糖对原料乳中微生物抑制作用的研究

通过研究壳寡糖对原料乳中细菌总数、嗜冷菌数、芽孢数及耐热芽孢数的抑制作用和不同浓度的壳寡糖对原料乳中美兰变色时间的影响,得出壳寡糖对原料乳中细菌总数和嗜冷菌数均有显著的抑制作用(P<0.05),对芽孢总数和耐热芽孢数没有显著的影响(P>0.05),并明显的延长原料乳中美兰的变色时间。