Phosphatidylethanolamine Deficiency Impairs Escherichia coli Adhesion by Downregulating Lipopolysaccharide Synthesis,Which is Reversible by High Galactose/Lactose Cultivation

As the initiation step of bacterial infection or biofouling, bacterial adhesion on cells or substrates is generally an optimal target for antibacterial design. Phosphatidylethanolamine (PE) is the principal phospholipid in bacteria, and its function in bacterial adhesion remains unclear. In this study, four E. coli strains including two PE-deficient mutants (PE^?PC^? and PE^?PC^+?strains) and two PE-containing wild-type controls (PE?⁺?PC^? strains) were recruited to investigate the influence of PE deficiency on bacterial adhesion. We found that PE deficiency could impair E. coli adhesion on macrophages (human THP-1-derived and mouse RAW264.7 macrophages) or glass coverslips by downregulating lipopolysaccharide (LPS) biosynthesis, which could be reversible by high galactose/lactose but not glucose cultivation. The data imply that PE play important role in bacterial adhesion probably via affecting LPS biosynthesis and suggest that targeting PE biosynthesis is also a potential antibacterial strategy.     

Taxoids profiling of suspension Taxus chinensis var. mairei cells in response to shear stress

To develop an optimal bioprocess for paclitaxel (Taxols) supply, taxoid biosynthetic pathway regulation must be better understood. The main taxoid metabolites (paclitaxel, baccatin III, taxol C, etc.) in Taxus cell culture showed great difference under shear stress. However, the regulating mechanism of taxoids metabolism under shear stress remained elusive. Here an efficient metabolic profiling approach combined with multivariate analysis was employed to profile taxoids changes of Taxus cells under laminar shear stress. A total of 21 taxoids were identified and quantified by ultra-performance liquid chromatography coupled with Q-TOF mass spectrometry. The result showed the contents of paclitaxel and baccatin III were reduced by shear stress, indicating the inhibitory effect of shear stress on paclitaxel biosynthesis. The levels of other taxoids uninvolved in paclitaxel biosynthesis were decreased except several metabolites. Further analysis of mapping measured taxoids concentrations onto paclitaxel biosynthesis pathway illustrating proposed intermediates and “off-pathway” metabolites revealed shear stress might disrupt the appropriate cyclization process of geranylgeranyl-pyrophosphate, aggravate the inappropriate order of hydroxylations and acylations, and not be good for functional group oxetane formation. These findings revealed the possible mechanism for shear stress limiting paclitaxel production and might have important biotechnological applications to increase the yields of paclitaxel and relevant precursors.     

基于扎根理论的社区参与国家公园建设与管理的机制研究

大量研究表明自然保护地周边社区居民对保护地的态度是影响社区参与保护,实现保护地管理目标的主要因素,由此建立了"认知-态度-参与"的行为逻辑。中国国家公园体制建设希望进一步推进社区与保护地的关系,全面提升自然保护管理效率,也建立在对这一行为逻辑的认同之上。为验证社区认知与行为的关系,并为促进不同利益相关者参与国家公园建设与管理提供科学支持,研究试图分析武夷山国家公园体制试点区社区居民的国家公园的"认知-态度-参与"的行为逻辑以揭示社区参与国家公园建设与管理的深层机制。与一般研究不同在于,本研究是针对正在建设中的国家公园,使得本地利益相关方的态度有机会影响国家公园建设进程。为获得社区"认知-态度-参与"核心观念,探索社区参与的过程和方式,提出促进社区参与的方法,研究采用扎根理论这一质性研究方法,基于对本地利益相关者采用半结构化开放问题的深度访谈,归纳出概念与范畴,形成社区参与国家公园建设与管理的理论。研究首先形成45个概念与13个范畴,通过范式模型研究发现,稳定资源使用权利、引导社区参与决策、保障社区发展权利、创新社区产业模式、提高个体适应能力、降低政策风险影响等六个主范畴能够解释社区居民对国家公园的认知-态度-参与这一行为逻辑。通过归纳主范畴,研究进一步提炼出社区参与国家公园建设与管理的理论核心在于社区自然资源管理的自主性、创新性与适应性,其内在机制在于社区参与是一个自主性提高的动态过程,需要通过保障社区生计资源禀赋来实现长期的价值转化,最终使其成为社会-生态系统适应性治理的组成部分。在理论阐释基础上,研究提出从改善社区认知、提升社区参与角度推进国家公园建设的路径。研究也指出,所提出的社区参与机制理论能够提炼量化指标评价社区参与,追踪武夷山国家公园体制试点进程。     

Continuous microalgae cultivation in a photobioreactor

New biomass sources for alternative fuels has become a subject of increasing importance as the nation strives to resolve the economic and strategic impacts of limited fossil fuel resources on our national security, environment, and global climate. Algae are among the most promising non‐food‐crop‐based biomass feedstocks. However, there are currently no commercially viable microalgae‐based production systems for biofuel production that have been developed, as limitations include less‐than optimal oil content, growth rates, and cultivation techniques. While batch studies are critical for determining basic growth phases and characteristics of the algal species, steady‐state studies are necessary to better understand and measure the specific growth parameters. This study evaluated the effects of dilution rate on microalgal biomass productivity, lipid content, and fatty acid profile under steady‐state conditions with continuous illumination and carbon dioxide supplemention for two types of algae. Continuous cultures were conducted for more that 3 months. Our results show that the productivity of Chlorella minutissima varied from 39 to 137 mg/L/day (dry mass) when the dilution rate varied from 0.08 to 0.64 day^?1. The biomass productivity of C. minutissima reached a maximum value (137 mg/L/day) at a dilution rate of 0.33 day^?1, while the productivity of Dunaliella tertiolecta varied from 46 to 91 mg/L/day at a dilution rate of 0.17 to 0.74 day^?1. The biomass productivity of D. tertiolecta reached a maximum value of 91 mg/L/day at a dilution rate of 0.42 day^?1. Moreover, the lipid content had no significant change with various dilution rates. Biotechnol. Bioeng. 2012; 109: 2468–2474. © 2012 Wiley Periodicals, Inc.     

Responses to ocean acidification and diurnal temperature variation in a commercially farmed seaweed,Pyropia haitanensis (Rhodophyta)

To investigate carbon and nitrogen metabolism in Pyropia haitanensis in response to the combined conditions of ocean acidification and diurnal temperature variation, maricultured thalli were tested in acidified culture under different temperature treatments. The results showed a combined effect of ocean acidification and diurnal temperature difference on the C and N metabolism and growth of P. haitanensis. In acidifed culture, algal growth, maximum photosynthetic rate, nitrate reductase (NR) activity, amino acid (AA) content and AA score (AAS) were more significantly enhanced in seaweed under diurnal temperature variation than in seaweed at constant temperature. In acidified seawater, soluble carbohydrates in P. haitanensis increased due to greater dissolved inorganic carbon (DIC), whereas soluble proteins decreased. Under the diurnal temperature treatment, higher temperature during the light period enhanced accumulation of algal photosynthates, whereas lower temperature in the dark period reduced energy consumption, resulting in enhanced algal growth, AA content and AAS. We concluded that suitable diurnal temperature difference would be conducive to C fixation and N assimilation under ocean acidification. However, excessively high temperatures would depress algal photosynthesis and increase energy consumption, thereby exerting a negative effect on algal growth.     

High-density culture of recombinant Chinese hamster ovary cells producing prothrombin in protein-free medium

A recombinant CHO cell line, CHO2DS, was immobilized on porous microcarrier Cytopore 1 and cultivated in 1 l modified Super-spinner and 2 l stirred tank bioreactor with the perfusion of a low-cost chemically defined protein-free medium DF6S. CHO2DS cells could enter into the inner space and grew both in the inner space and on the surface of Cytopore 1 in DF6S and produced prothrombin at 22 mg l^–1 after 10 days. From a seeding density of 5.7 × 10⁵ cells ml^–1, the highest viable cell density of CHO2DS was 1.12 × 10⁷ cells ml^–1.     

Protected raspberry production alters aphid–plant interactions but not aphid population size

• 1 Aphid population dynamics in crops are often driven by interactions with their host plants, which can be extensively influenced by environmental change. Protective environments (i.e. plastic tunnels) are now frequently used for soft fruit production, which may affect the localized climate and alter such interactions. This two year study on red raspberry (Rubus idaeus) addressed how protected environments affected two aphid species; the large raspberry aphid Amphorophora idaei (LRA) and the small raspberry aphid Aphis idaei (SRA).
• 2 Temperatures were higher (up to 7–10 °C) in tunnels compared with the field. Plants in tunnels grew approximately 1.4 cm/week faster and had lower (approximately 35%) foliar amino acid concentrations than plants in the field.
• 3 Aphids affected plant growth differently depending on growing environment; they promoted plant growth by 18–37% in tunnels, although they had no such effect in the field. Aphids reduced total and essential amino acid concentrations, with SRA causing greatest reductions (approximately 40% and 33%, respectively).
• 4 Aphid population sizes were similar in both environments, although individual LRA were smaller in tunnels (30% smaller in 2007) compared with those in the field. We suggest that faster aphid development rates inside warmer tunnels were not realized as a result of the variable effects of the growing environment on amino acid composition.
• 5 We conclude that the increasing use of protected environments in crop production will not necessarily cause predictable increases in aphid populations, although it may alter aphid–plant interactions in terms of aphid‐induced changes to plant growth.

     

Galactose-limited fed-batch cultivation of Escherichia coli for the production of lacto-N-tetraose

Lacto-N-tetraose (Gal(β1-3)GlcNAc(β1-3)Gal(β1-4)Glc) is one of the most abundant oligosaccharide structures in human milk. We recently described the synthesis of lacto-N-tetraose by a whole-cell biotransformation with recombinant Escherichia coli cells. However, only about 5% of the lactose was converted into lacto-N-tetraose by this approach. The major product obtained was the intermediate lacto-N-triose II (GlcNAc(β1-3)Gal(β1-4)Glc).In order to improve the bioconversion of lactose to lacto-N-tetraose, we have investigated the influence of the carbon source on the formation of lacto-N-tetraose and on the intracellular availability of the glycosyltransferase substrates, UDP-N-acetylglucosamine and UDP-galactose. By growth of the recombinant E. coli cells on D-galactose, the yield of lacto-N-tetraose (810.8 mg L⁻¹ culture) was 3.6-times higher compared to cultivation on D-glucose.Using fed-batch cultivation with galactose as sole energy and carbon source, a large-scale synthesis of lacto-N-tetraose was demonstrated. During the 26 h feeding phase the growth rate (μ = 0.05) was maintained by an exponential galactose feed. In total, 16 g L⁻¹ lactose were fed and resulted in final yields of 12.72 ± 0.21 g L⁻¹ lacto-N-tetraose and 13.70 ± 0.10 g L⁻¹ lacto-N-triose II. In total, 173 g of lacto-N-tetraose were produced with a space-time yield of 0.37 g L⁻¹ h⁻¹.