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.     

大豆功能性成分的开发现状

大豆是我国的一种传统食品,含有丰富的生理活性物质,包括有大豆蛋白质、大豆异黄酮、大豆皂苷、大豆低聚糖以及大豆膳食纤维等,此外大豆发酵制品还会产生一些生理活性成分如蛋白黑素、大豆多肽等。综述目前大豆功能性成分的开发现状,同时也介绍了它们所具有的相关生理功能。     

Engineered Glycosidases for the Synthesis of Analogs of Human Milk Oligosaccharides

Enzymatic synthesis is an elegant biocompatible approach to complex compounds such as human milk oligosaccharides (HMOs). These compounds are vital for healthy neonatal development with a positive impact on the immune system. Although HMOs may be prepared by glycosyltransferases, this pathway is often complicated by the high price of sugar nucleotides, stringent substrate specificity, and low enzyme stability. Engineered glycosidases (EC 3.2.1) represent a good synthetic alternative, especially if variations in the substrate structure are desired. Site-directed mutagenesis can improve the synthetic process with higher yields and/or increased reaction selectivity. So far, the synthesis of human milk oligosaccharides by glycosidases has mostly been limited to analytical reactions with mass spectrometry detection. The present work reveals the potential of a library of engineered glycosidases in the preparative synthesis of three tetrasaccharides derived from lacto-N-tetraose (Galβ4GlcNAcβ3Galβ4Glc), employing sequential cascade reactions catalyzed by β3-N-acetylhexosaminidase BbhI from Bifidobacterium bifidum, β4-galactosidase BgaD-B from Bacillus circulans, β4-N-acetylgalactosaminidase from Talaromyces flavus, and β3-galactosynthase BgaC from B. circulans. The reaction products were isolated and structurally characterized. This work expands the insight into the multi-step catalysis by glycosidases and shows the path to modified derivatives of complex carbohydrates that cannot be prepared by standard glycosyltransferase methods.     

从豆清水中提取大豆低聚糖的工艺研究

介绍了从豆清水中提取大豆低聚糖的工业化生产工艺,并对成品大豆低聚糖的基本性质进行了研究.实验选择高温离心分离法脱除豆清水中的蛋白质,温度选择90℃,脱除率达85.12％.选择电渗析法进行脱盐处理,脱除率达85.8％.使用粉末活性炭进一步进行脱色,采用单因素及正交实验对脱色工艺进行研究,最佳脱色工艺为活性炭用量0.3％,40℃下脱色35min,脱色率达83.71％,低聚糖保留率为80.41％.     

发酵法生产魔芋葡甘聚糖酶的研究

目的：探索发酵生产葡甘聚糖酶的最佳条件。方法：在摇瓶培养的基础上，比较不同温度、pH和接种量对发酵反应的影响，并在5L发酵罐上用正交试验筛选最佳的产酶条件配伍。结果：该菌产生葡甘聚糖酶的最佳条件为温度50℃，pH5．5～6．0，接种量10％，发酵前期搅拌速度50rpm，通气量20L／h，6h后搅拌速度改为100rpm，通气量10L／h，在这个条件下发酵获得的酶比活力为4812U／mg，总活力达到7810U／mL。培养8h后细菌生物量、产物含量迅速提高，24h达到顶峰时期。发酵动力学属于偶联型。     

Recent advances refining galactooligosaccharide production from lactose

The production of prebiotic galactooligosaccharides (GOS) by the β-galactosidase catalysed conversion of lactose has become commercially important. Yet it remains a challenge to sufficiently understand the structure and activity of β-galactosidase, to increase the efficiency of transgalactosylation and GOS production and to improve the quality of GOS products in a rational way. This review covers the broad but related aspects of GOS synthesis including: the structure and reaction mechanism of β-galactosidase, factors effecting yield and productivity of GOS synthesis systems, the structure of GOS products, models for the kinetics of GOS synthesis and reactor configurations for GOS synthesis. It aims to couple recent discoveries with established knowledge to enhance understanding of the complex biochemistry of GOS synthesis.     

功能性低聚糖--水苏糖

水苏糖作为一种新兴的功能性低聚糖类 ,其开发和应用日益受到重视。本文介绍了水苏糖的性质、生理功能、制备方法及应用 ,并对其开发应用前景进行了展望。