吡咯喹啉醌合成及生产工艺研究进展

吡咯喹啉醌(pyrroloquinoline quinone, PQQ)是继烟酰胺和核黄素之后发现的第三类氧化还原酶辅因子,普遍存在于生物体中参与呼吸链电子传递,具有促进线粒体产生、清除自由基、增强细胞代谢和预防心肌损伤等生理功能,在医药、食品和农业领域具有广泛的应用前景。微生物发酵法是PQQ生产的主要方式,解析PQQ生物合成途径及其调控机制,通过代谢工程选育短周期、高产量的生产菌是PQQ工业化的研究方向之一。本文综述了PQQ的合成途径、高产菌株选育以及微生物发酵生产与分离纯化的研发工作,为深入阐释PQQ的生物合成机制和工业化生产菌株的选育提供参考。     

RESPIRATORY EFFECTS UPON THE VISUAL THRESHOLD

Measurements are reported of the effects of respiratory stresses upon the absolute threshold of peripheral (rod) vision. Since subjects were kept wholly dark adapted and the photochemical system of the rods therefore stationary, the changes recorded may be assumed to have originated more centrally. To this degree the measurements provide a quantitative index of central nervous imbalance. Breathing room air or 32 to 36 per cent oxygen at about double the normal rate causes the visual threshold to fall to approximately half the normal value within 5 to 10 minutes. This change is due primarily to alkalosis induced by the hyperventilation, and can be abolished or reversed by adding carbon dioxide to the inspired mixtures. Normal or rapid breathing of 2 per cent carbon dioxide causes no change in threshold; with 5 per cent carbon dioxide the threshold is approximately doubled. Breathing 10 per cent oxygen at the normal rate also approximately doubles the threshold. This effect is compensated in part by rapid breathing. When 10 per cent oxygen is breathed at twice the normal rate the threshold usually falls at first, then slowly rises to supernormal levels. Due primarily to variations in their breathing patterns subjects yield characteristically different responses on sudden exposure to low oxygen tensions with breathing uncontrolled. The threshold may either rise or fall; and on release from anoxia it may rise, or fall to normal or subnormal levels. The threshold adjusts to anoxia rapidly; exposures lasting 5 to 6 hours do not produce greater or more persistent changes than those of much shorter duration.     

Assembly Kinetics of Vimentin Tetramers to Unit-Length Filaments: A Stopped-Flow Study

Intermediate filaments (IFs) are principal components of the cytoskeleton, a dynamic integrated system of structural proteins that provides the functional architecture of metazoan cells. They are major contributors to the elasticity of cells and tissues due to their high mechanical stability and intrinsic flexibility. The basic building block for the assembly of IFs is a rod-like, 60-nm-long tetrameric complex made from two antiparallel, half-staggered coiled coils. In low ionic strength, tetramers form stable complexes that rapidly assemble into filaments upon raising the ionic strength. The first assembly products, “frozen” by instantaneous chemical fixation and viewed by electron microscopy, are 60-nm-long “unit-length” filaments (ULFs) that apparently form by lateral in-register association of tetramers. ULFs are the active elements of IF growth, undergoing longitudinal end-to-end annealing with one another and with growing filaments. Originally, we have employed quantitative time-lapse atomic force and electron microscopy to analyze the kinetics of vimentin-filament assembly starting from a few seconds to several hours. To obtain detailed quantitative insight into the productive reactions that drive ULF formation, we now introduce a “stopped-flow” approach in combination with static light-scattering measurements. Thereby, we determine the basic rate constants for lateral assembly of tetramers to ULFs. Processing of the recorded data by a global fitting procedure enables us to describe the hierarchical steps of IF formation. Specifically, we propose that tetramers are consumed within milliseconds to yield octamers that are obligatory intermediates toward ULF formation. Although the interaction of tetramers is diffusion controlled, it is strongly driven by their geometry to mediate effective subunit targeting. Importantly, our model conclusively reflects the previously described occurrence of polymorphic ULF and mature filaments in terms of their number of tetramers per cross section.     

Random Motion of Chromatin Is Influenced by Lamin A Interconnections

Using fluorescence correlation spectroscopy in single-plane illumination microscopy, we investigated the dynamics of chromatin in interphase mouse adult fibroblast cell nuclei under the influence of the intermediate filament protein lamin A. We find that 1) lamin A-eGFP and histone H2A-mRFP show significant comobility, indicating that their motions are clearly interconnected in the nucleus, and 2) that the random motion of histones H2A within the chromatin network is subdiffusive, i.e., the effective diffusion coefficient decreases for slow timescales. Knocking out lamin A changes the diffusion back to normal. Thus, lamin A influences the dynamics of the entire chromatin network. Our conclusion is that lamin A plays a central role in determining the viscoelasticity of the chromatin network and helping to maintain local ordering of interphase chromosomes.     

Screening and identification of a microsatellite marker associated with sex in Wami tilapia, <Emphasis Type="Italic">Oreochromis urolepis hornorum</Emphasis>

In this study, primer pairs of 15 microsatellite markers associated with sex determination of tilapia were selected and amplified in Wami tilapia, Oreochromis urolepis hornorum. While one marker, UNH168, on linkage group 3 (LG3) was associated (P < 0.001) with the phenotypic sex in the experimental population, nine genotypes were detected in both sexes. Only 99-bp allele was detected in the female samples, while 141, 149 and 157-bp alleles were present in both male and female samples. UNH168 was localized by fluorescence in situ hybridization (FISH) on the long arm of the largest tilapia chromosome pair (chromosome 1, equivalent to LG3). This sex-linked microsatellite marker could potentially be used for marker-assisted selection in tilapia breeding programmes to produce monosex male tilapia.     

NaCl胁迫对甘薯叶片叶绿体超微结构及一些酶活性的影响

随ＮａＣｌ 胁迫浓度的提高,甘薯叶片叶绿体数目逐渐减少, 类囊体膜片层松散、扭曲、破裂并逐渐解体, 叶绿素含量下降。与此同时,Ｈ２Ｏ２ 、ＭＤＡ 含量增加, ＡＳＰ、ＳＯＤ 活性表现出先上升后下降的趋势。耐盐品种在ＮａＣｌ 胁迫下能维持较强的Ｈ２Ｏ２ 清除能力和较低的ＭＤＡ 水平