Synthesis of L-Rhamnose and N -Acetyl-D-Glucosamine Derivatives Entering in the Composition of Bacterial Polysaccharides by Use of Glucansucrases

Transglucosylation reactions using sucrose as glucosyl donor and either N-acetyl-D-glucosamine, L-rhamnose, or methyl α -L-rhamnopyranoside as acceptors were carried out with recombinant glucansucrases from families 70 and 13 of glycoside-hydrolases. Depending on the enzyme specificity, various carbohydrate structures were synthesized and characterized including α -D-glucopyranosyl-(1 → 6)-N-acetyl-D-glucosamine, α -D-glucopyranosyl-(1 → 4)-N-acetyl-D-glucosamine, α -D-glucopyranosyl-(1 → 1)-β -L-rhamnopyranoside, α -D-glucopyranosyl-(1 → 4)-α -D-glucopyranosyl-(1 → 1)-β -L-rhamnopyranoside, methyl α -D-glucopyranosyl-(1 → 4)-α -L-rhamnopyranoside, and methyl α -D-glucopyranosyl-(1 → 3)-α -L-rhamnopyranoside. Disaccharides were obtained with yields going up to 64%. The structural diversity generated as well as the obtained yields appear to be related to enzyme active site architecture, which can be modulated and improved by enzyme engineering. Several of the obtained disaccharides enter in the composition of surface polysaccharides of pathogenic bacteria, among which is Shigella flexneri. Our results outline the potential of glucansucrases in the chemo-enzymatic synthesis of complex carbohydrates of biological interest whose chemical synthesis may be seen as a limitation.     

细菌纤维素纳米纤维稳定乳液的性能

采用超声和高压均质两种方式分散的细菌纤维素(BC)悬浮液制备了BC纳米纤维稳定的水包油型Pickering乳液, 并考察了纤维用量、 pH值和机械分散方式对乳液稳定性的影响. 结果表明, 乳液的稳定性随纳米纤维用量的增加而增加; 碱性条件比酸性条件制备的乳液稳定性高, 且在pH=12时达到最高. 用高压均质方式分散的BC稳定乳液的效果优于采用超声方式分散的BC的效果, 这是由于高压均质后的纤维较短, 可以提供更多的纳米纤维稳定乳液. 计算结果表明, BC纳米纤维在液体石蜡/水界面上的三相接触角为72.5°, 说明BC适合稳定水包油型乳液.     

Stereochemical differentiation and simultaneous analysis of 3-, 4-, and 5-hydroxyalkanoic acids from biopolyesters by multidimensional gas chromatography

The direct enantioselective analysis of 3-, 4-, and 5-hydroxy fatty acids from biological material has been achieved by enantioselective multidimensional gas chromatography (enantio-MDGC) with heptakis(2,3-di-O-methyl-6-O-tert-butyldimethylsilyl)- or (2,3-di-O-acetyl-6-O-tert-butyldimethylsilyl)-β-cyclodextrin as chiral stationary phase. All the bacteria investigated produced polyesters of enatiomerically pure (R) configured compounds.     

细菌纤维素(BC)增强PVA/PVP复合水凝胶的制备及性能表征

通过冷冻-熔融法制备了细菌纤维素/聚乙烯醇/聚乙烯吡咯烷酮(BC/PVA/PVP)双网络复合水凝胶,并采用X射线衍射,红外光谱,扫描电镜,力学性能测试等手段对凝胶的结构和性能进行表征.研究发现PVA、PVP通过氢键作用均匀地吸附于纤维微丝周围,将BC纤维有效地分开,因而干燥后的复合凝胶在热水中浸泡后仍可恢复原状;X射线...     

Electrochemical attachment of motile bacterial cells to gold

Selective attachment of Escherichia coli K-12 bacterial cells to charged gold surfaces was demonstrated. Electrostatic binding of E. coli K-12 bacterial cells to positively charged surfaces was observed starting at +750 mV. The binding of E. coli K-12 cells to positively charged gold surfaces is proposed to occur due to long-range electrostatic interactions between the negatively charged O-chain of lipopolysaccharide (LPS) molecules protruding the bacterial cell body and the electrode surface. Removing LPS alters the cellular surface charge and results in cellular attachment to negatively charged surfaces. Thus, applying an electrical potential allows for the direct, real time detection of live, dead or damaged bacterial cells. The attachment of E. coli K-12 bacterial cells to surfaces with an applied potential substantiates the hypothesis that an electrostatic interaction is responsible for the binding of bacterial cells to positively charged molecular assemblies on surfaces used for building bacterial microarrays.     

铜锌超氧化物歧化酶的突变与神经退行性紊乱的生物无机化学

本文简单介绍了铜锌超氧化物歧化酶的结构、功能及其相互关系,较为详细地讨论了与神经退行性紊乱,尤其是与肌萎性脊髓侧索硬化症有关的铜锌超氧化物歧化酶突变体的生物无机化学的研究进展,提出由该酶突变而引起的蛋白质局部结构变化和金属离子缺失,以及由此导致的氧化损伤和聚集作用可能是导致神经退行性紊乱的主要原因之一.     

Application of horizontal staircase electrophoresis in agarose minigels to the random intergenic spacer analysis of clinical samples

The random intergenic spacer analysis is a recently developed technique for the study of microbial populations. The bacterial intergenic spacer (ITS) is located between 16S rRNA and 23S rRNA genes and presents different length and sequence among bacterial species. Therefore, the amplicons can be separated by electrophoresis commonly performed at low voltage during several hours. Although this technique is especially useful for unculturable microorganisms, it has not been applied before to clinical sample analysis. As these samples have a limited number of bacterial species, the size of the gels may be reduced to facilitate their handling and to reduce the running time. To obtain maximum separation among the ITS bands, we analysed in this work different electrophoretical conditions including staircase electrophoresis, a technique based on the application of several voltage steps. The results obtained showed a different behaviour of the electrical resistance during the performance of submarine horizontal and vertical staircase electrophoresis. In the first case the resistance decreased during most of the running time whereas in the second case it increased. Here, we show that the performance of horizontal staircase electrophoresis reduces the running time more than 80% with respect to conventional electrophoresis at low voltages. This procedure was applied to the separation of ITS bands from bacterial DNA present in a tissue from a vocal cord biopsy. The sequencing of these bands allowed their identification. This new procedure may be very useful in the rapid diagnosis of bacteria present in human, animal and plant tissues.     

Immobilized bacterial luciferase and its applications

This review discusses the properties of the bioluminescent bacterial system as well as the methods for immobilization of bacterial luciferases and for their co-immobilization with other enzymes. The analytical systems using immobilized bacterial luciferases and their applications in analytical biochemistry and biotechnology have been described.     

Improvement in mechanical properties and biocompatibility of biosynthetic bacterial cellulose/lotus root starch composites

Bacterial cellulose/lotus root starch (BC/LRS) composites were prepared by cultivating Acetobacter xylinum in nutrient media containing gelatinized lotus root starch. Low concentrations of gelatinized LRS had increased BC production with the maximum value at 6.67 g/L when 5 g/L of LRS was added in the culture media and the composites had thicker and denser fibrils compared with those of BC with low concentrations of LRS (2.5 and 5 g/L). When the concentration of LRS was increased above 7.5 g/L, the morphology of the BC/LRS composites contained more fibril layers that were linked with LRS. The results from X-ray diffraction (XRD) demonstrated that there was no significant difference in structure between BC and BC/LRS composites except a slight increase in crystallinity for BC/LRS composites as the concentration of LRS was lifted up. The tensile tests were performed to display BC/LRS composites prepared with LRS concentration at 2.5 and 5 g/L in media had the tensile strength of 54 and 60 MPa, respectively, which indicated an improvement in mechanical property compared to the unmodified BC (45 MPa). Live/dead assay with chondrocytes seeded on BC/LRS composite revealed higher cell viability ranging from 85% to 90% than BC. Furthermore, cell morphology with typical spindle shape was observed on the surfaces of BC/LRS composite by confocal microscope. Through the overall results, it shows that this study has provided a guidance to prepare BC/LRS composites with better cell biocompatibility and higher mechanical strength than those of BC for the potential use in cartilage tissue engineering.