Fructose uptake in Bifidobacterium longum NCC2705 is mediated by an ATP-binding cassette transporter

Recently, a putative ATP-binding cassette (ABC) transport system was identified in Bifidobacterium longum NCC2705 that is highly up-regulated during growth on fructose as the sole carbon source. Cloning and expression of the corresponding ORFs (bl0033-0036) result in efficient fructose uptake by bacteria. Sequence analysis reveals high similarity to typical ABC transport systems and suggests that these genes are organized as an operon. Expression of FruE is induced by fructose, ribose, or xylose and is able to bind these sugars with fructose as the preferred substrate. Our data suggest that BL0033-0036 constitute a high affinity fructose-specific ABC transporter of B. longum NCC2705. We thus suggest to rename the coding genes to fruEKFG and the corresponding proteins to FruE (sugar-binding protein), FruK (ATPase subunit), FruF, and FruG (membrane permeases). Furthermore, protein-protein interactions between the components of the transporter complex were determined by GST pulldown and Western blot analysis. This revealed interactions between the membrane subunits FruF and FruG with FruE, which in vivo is located on the external side of the membrane, and with the cytoplasmatic ATPase FruK. This is in line with the proposed model for bacterial ABC sugar transporters.     

Plasma half-life extension of small recombinant antibodies by fusion to immunoglobulin-binding domains

Many therapeutic proteins possessing a small size are rapidly cleared from circulation. Half-life extension strategies have therefore become increasingly important to improve the pharmacokinetic and pharmacodynamic properties of protein therapeutics. Here, we performed a comparative analysis of the half-life extension properties of various bacterial immunoglobulin-binding domains (IgBDs) derived from Staphylococcus protein A (SpA), Streptococcus protein G (SpG), and Finegoldia (formerly Peptostreptococcus) protein L (PpL). These domains, composed of 50-60 amino acid residues, were fused to the C terminus of a single-chain Fv and a bispecific single-chain diabody, respectively. All fusion proteins were produced in mammalian cells and retained their antigen-binding properties. The half-lives of the antibody molecules were prolonged to varying extents for the different IgBDs. The strongest effects in mice were observed for domain C3 of SpG (SpG(C3)) followed by domains B and D of SpA, suggesting that SpG(C3) is particularly useful to extend the plasma half-life of small proteins.     

Genetic Engineering of the Phosphocarrier Protein NPr of the Escherichia coli Phosphotransferase System Selectively Improves Sugar Uptake Activity

The Escherichia coli phosphoenolpyruvate:sugar phosphotransferase system (PTS) in prokaryotes mediates the uptake and phosphorylation of its numerous substrates through a phosphoryl transfer chain where a phosphoryl transfer protein, HPr, transfers its phosphoryl group to any of several sugar-specific Enzyme IIA proteins in preparation for sugar transport. A phosphoryl transfer protein of the PTS, NPr, homologous to HPr, functions to regulate nitrogen metabolism and shows virtually no enzymatic cross-reactivity with HPr. Here we describe the genetic engineering of a "chimeric" HPr/NPr protein, termed CPr14 because 14 amino acid residues of the interface were replaced. CPr14 shows decreased activity with most PTS permeases relative to HPr, but increases activity with the broad specificity mannose permease. The results lead to the proposal that HPr is not optimal for most PTS permeases but instead represents a compromise with suboptimal activity for most PTS permeases. The evolutionary implications are discussed.     

Physicochemical and sorption properties of thermally-treated sediments with high organic matter content

Sediment samples with high organic carbon contents (22.04% and 8.46%) were collected and thermally-treated using a method analogous to biochar production. The obtained thermally-treated sediments (TTSs) showed a much higher degree of carbon capture in comparison to biochar derived from common biomass, indicating potential use of TTSs in soil amendment and carbon sequestration. Their sorption with organic contaminants was also investigated using sulfamethoxazole (SMX) as a model sorbate. SMX sorption increased greatly with pyrolytic temperature. Desorption ratio of the adsorbed SMX in TTSs generally decreased with increased pyrolytic temperature and with decreased solid-phase concentrations. The thermodynamic analysis showed that the higher entropy increase (positive ΔS) was well related with the decreased desorption ratio with increased solid-phase concentration for the original sediments. The fate-controlling effect of contaminants in TTS application for soil amendment should be evaluated combining sorption/desorption and sorption thermodynamic studies.     

滇池北部湖区浮游植物时空格局及相关环境因子

为研究滇池北部湖区浮游植物的时空格局,探讨影响格局形成的关键影响因子,2008年12月至2009年6月,在滇池北部湖区选择6个点位,共开展了24次采样,调查并分析了浮游植物群落及其相关理化参数。通过比对滇池全湖周年监测数据,研究发现,北部湖区叶绿素a浓度从12月中旬开始下降,2月中旬降至最低值26.5μg/L,5月中旬达到最大值530.5μg/L。调查期间共鉴定出浮游植物74属97种,其中绿藻53种,蓝藻20种,硅藻17种,隐藻2种,金藻2种,甲藻1种,裸藻2种。蓝藻生物量在12月、3月至6月期间占优势,12月份优势种为绿色微囊藻(Microcystis viridis),3月至5月为水华束丝藻(Aphanizomenonflos-aquae),6月为惠氏微囊藻(Microcystis wesenbergii)。硅藻在1月占优势,优势种为颗粒直链藻最窄变种(Melosira granulata var.angustissima)。研究结果表明,水温高于14℃以上,微囊藻生物量迅速增加;束丝藻生物量随温度增加而增加,高于22℃时迅速减少。北部湖区6个点位浮游植物的空间差异显著,D1、D4号点微囊藻、束丝藻生物量显著高于其他四个点(P<0.01),D2、D5号点广缘小环藻生物量显著高于其他四个点(P<0.01)。研究结果显示,在富营养化严重的滇池北部湖区,水温可能为影响浮游植物种类组成及生物量的关键因子,浮游植物水平分布的差异主要由气象因素和水体营养盐共同决定。     

以工程素质培养为导向的生物工程专业核心课程改革

探讨以注重综合素质、强化工程能力和工程素质的培养、完善知识结构为重点的生物工程专业核心课程的教学改革。阐明生物工程师应具备的以五种能力和五种意识为核心的工程素质内涵,明确培养工程素质实施的核心课程;充分利用产学研平台,建设素质精良的"双师型"教师队伍;以工程能力培养为主线,建立以生物工厂设计和工艺改进的教学特色课程群;实施启发式、分组讨论法、比较归纳法和案例式的教学方法改革,提高了教学效果,强化了生物工程专业学生工程素质的培养。     

丛毛单胞菌在环境污染物降解方面的研究进展

环境污染物的微生物降解是环保领域研究的一个热点。研究发现,丛毛单胞菌对多种污染物有很好的降解效果。综述目前已分离的一些丛毛单胞菌对不同污染物的降解特性、代谢途径,并分析了影响其降解效果的各种因素,介绍应用现状并对前景进行展望。     

Microbial diversity and composition of the sediment in the drinking water reservoir Saidenbach (Saxonia, Germany)

Sediments contain a huge number and diversity of microorganisms that are important for the flux of material and are pivotal to all major biogeochemical cycles. Sediments of reservoirs are affected by a wide spectrum of allochthous and autochthonous influences providing versatile environments along the flow of water within the reservoir. Here we report on the microbial diversity in sediments of the mesotrophic drinking water reservoir Saidenbach, Germany, featuring a pronounced longitudinal gradient in sediment composition in the reservoir system. Three sampling sites were selected along the gradient, and the microbial communities in two sediment depths were characterized using catalysed reporter deposition fluorescence in situ hybridization (CARD-FISH) and a bar-coded pyrosequencing approach. Multivariate statistic was used to reveal relationships between sequence diversity and the environmental conditions. The microbial communities were tremendously diverse with a Shannon index of diversity (H') ranging from 6.7 to 7.1. 18,986 sequences could be classified into 37 phyla including candidate divisions, but the full extent of genetic diversity was not captured. While CARD-FISH gave an overview about the community composition, more detailed information was gained by pyrosequencing. Bacteria were more abundant than Archaea. The dominating phylum in all samples was Proteobacteria, especially Betaproteobacteria and Deltaproteobacteria. Furthermore, sequences of Bacteroidetes, Verrucomicrobia, Acidobacteria, Chlorobi, Nitrospira, Spirochaetes, Gammaproteobacteria, Alphaproteobacteria, Chloroflexi, and Gemmatimonadetes were found. The site ammonium concentration, water content and organic matter content revealed to be strongest environmental predictors explaining the observed significant differences in the community composition between sampling sites.     

Recent advances in two-phase partitioning bioreactors for the treatment of volatile organic compounds

Biological processes are considered to be the most cost-effective technology for the off-gas treatment of volatile organic compounds (VOC) at low concentrations. Two-phase partitioning bioreactors (TPPBs) emerged in the early 1990s as innovative multiphase systems capable of overcoming some of the key limitations of traditional biological technologies such as the low mass transfer rates of hydrophobic VOCs and microbial inhibition at high VOC loading rates. Intensive research carried out in the last 5 years has helped to provide a better understanding of the mass transfer phenomena and VOC uptake mechanisms in TPPBs, which has significantly improved the VOC biodegradation processes utilizing this technology platform. This work presents an updated state-of-the-art review on the advances of TPPB technology for air pollution control. The most recent insights regarding non-aqueous phase (NAP) selection, microbiology, reactor design, mathematical modeling and case studies are critically reviewed and discussed. Finally, the key research issues required to move towards the development of efficient and stable full-scale VOC biodegradation processes in TPPBs are identified.     

The soft-bottom macrobenthos of Gray's Reef National Marine Sanctuary and nearby shelf waters off the coast of Georgia, USA

As part of an ongoing ecological assessment of the Gray's Reef National Marine Sanctuary (GRNMS), a 58-km² marine protected area 32 km off the coast of Georgia, USA, surveys of benthic macroinfaunal communities, contaminant levels in sediments and biota, and general habitat conditions were conducted during 2000-2002 at 20 stations within the sanctuary and along three cross-shelf transects in nearby shelf waters. Macroinfaunal community structure and composition exhibited distinct cross-shelf patterns associated with sediment granulometry, depth and possibly other factors related to shoreline proximity (e.g., erosional effects, recruitment of estuarine species). Finer-scale spatial patterns of benthic fauna among stations within the sanctuary appear to be related to proximity to live-bottom habitat and other features of seafloor structure (e.g., rippled vs. flat sand). Population densities of dominant fauna within the sanctuary also varied considerably among years, resulting in shifts in the ranking of dominants at most stations. Chemical contaminants generally were at low background concentrations below probable bioeffect levels and thus are not a likely cause of the observed spatial patterns of benthic fauna. However, trace concentrations of pesticides, PCBs, and PAHs were detectable in sediments and biota throughout the study area, demonstrating that chemicals originating from human activities are capable of reaching the offshore sanctuary environment, possibly from atmospheric deposition or cross-shelf transport of materials outwelled through coastal sounds. Highly diverse infaunal assemblages also were observed within the sanctuary and nearby sites of similar depth, suggesting that the sanctuary is an important reservoir of marine biodiversity. Results of this study should be useful in addressing long-term science and management needs of the GRNMS and in furthering our understanding of broader ecological patterns and dynamics of the surrounding South Atlantic Bight (SAB) ecosystem.