Engineering aspects of immobilized lipases on esterification: A special emphasis of crowding,confinement and diffusion effects

Cross‐linked enzyme crystal (CLEC) and sol‐gel entrapped pseudomonas sp. lipase were investigated for the esterification of lauric acid with ethanol by considering the effects of reaction conditions on reaction rate. The activation energy for the reaction was estimated to be 1097.58 J/mol and 181.75 J/mol for sol‐gel and CLEC entrapped lipase respectively. CLEC lipase exhibited a marginal internal diffusion effect on reaction rate over sol‐gel lipases and found to be interesting. The overall reaction mechanism was found to conform to the Ping Pong Bi Bi mechanism. The higher efficiency of sol‐gel lipases over CLEC lipases in esterification reaction is mainly due to the combined effects of crowding, confinement and diffusional limitations.     

Atomistic simulation of the sol formation during synthesis of organic/inorganic hybrid materials

Atomistic modelling was employed to investigate interactions between the precursors used in preparation of organic/inorganic polymer hybrids. Three molecular systems composed of different kinds of organofunctionalised silanes and an organic polymer in aqueous solution have been simulated, representing various model stages of the hydrolysis reaction during the sol/gel processing. The equilibrium configurations obtained by Molecular Dynamics were analysed in order to elucidate the extent of the hydrogen bonding network and clustering of components within the systems. Diffusion coefficients were obtained to estimate the mobility of the components and the stability of the molecular associations. It has been concluded that the molecular mechanism of the initial stages of the sol/gel process used for preparation of the organic/inorganic hybrids involves clustering of the organofunctionalysed inorganic components as a first step and then progressive hydrolysis as consequent steps. The organic polymer component plays a role of the cluster "wrap" and thus prevents the particles from unlimited growth and precipitation from the solution.     

聚赖氨酸溴化氢结合物的表面增强拉曼光谱

银溶胶对聚赖氨酸溴化氢结合物(PLys—HBr)表现出极大表面增强拉曼(SER)效应。同PLyS—HBr的普通拉曼光谱相比,表面增强因子提高达6个数量级。实验表明,NH_3~+基是银表面增强效应的强活性基团。但是在相同条件下,聚谷氨酸钠(PGA—Na)在银溶胶中不能获得SER光谱,这可能是由于空间障碍或者COO~-基的活性不如NH_3~+基。     

Glucose‐6‐phosphate dehydrogenase encapsulated in silica‐based hydrogels for operation in a microreactor

The future of hydrogen as fuel strongly depends on the possibility to produce it in an economic and clean way. Hydrogen can be produced from carbohydrates and water under mild conditions by means of a multistep synthetic pathway (13 enzymes) with very high yield. Crossover inhibitions and different optimal conditions of involved enzymes hinder the use of one‐pot approach. Immobilization of enzymes in coupled individual reactors may avoid this problem. This work deals with the immobilization in silica‐based hydrogels of one key enzyme of this pathway: glucose 6‐phosphate dehydrogenase from Leuconostoc mesenteroides. The carriers were prepared with an ethylene glycol‐modified silane, two polymers (polyethylene oxide and Pluronic®) and amino groups created by 3‐aminopropyltriethoxysilane. These parameters influenced the enzymatic activity after immobilization. Gels prepared by addition of polyethylene oxide gave the best results and were used as monoliths in microreactors with two different geometries. The systems showed a high operational stability but a low effective enzyme activity. Enzyme leaching and a nonideal flow pattern may account for the low activity observed. This work is possibly the first one dealing with the immobilization of glucose 6‐phosphate dehydrogenase in silica‐based gels for its application in flow‐through microreactors.     

1H NMR studies: dynamics of water in gelatin

 Proton magnetic resonance was used to characterize the dynamics of water in gelatin. Both sol and gel states were investigated. Transverse relaxation rates (R ₂) were dependent on the proton frequency measurement. (R ₂) measured with the Carr-Purcell-Meiboom-Gill pulse sequence was dependent on pulse spacing. These observations were interpreted in terms of chemical exchanges between water protons and those of the macromolecules in the sol state, whereas in the gel state the contribution of diffusion through microheterogeneities in the sample seems to provide an additional transverse relaxation mechanism. Received: 10 May 1999 / Revised version: 13 December 1999 / Accepted: 25 January 2000     

Differential and independent manifestation within co-containing neurones of neuropeptide Y and tyrosine hydroxylase during ontogeny of the rat central nervous system

The development of tyrosine hydroxylase- and neuropeptide Y-immunoreactive cell bodies in the foetal rat brain was analyzed immunohistochemically using antibodies raised against tyrosine hydroxylase and neuropeptide Y. Possible co-existence of these two substance within the same neurones was investigated by comparison of adjacent sections.

In the ventral medulla oblongata, neurones containing both neuropeptide Y- and tyrosine hydroxylase-like immunoreactivity were demonstrable in and around the lateral reticular nucleus as early as the 17th day of gestation. The total number and the proportion of cells exhibiting this co-existence increased from this stage up to birth. In the nucleus of the solitary tract in the dorsal medulla oblongata, NPY-immunoreactive cells bodies were first visualized at day 13 of gestation. However, although tyrosine hydroxylase-immunoreactive cells could also be seen within the nucleus at this and later ages, they occupied a different, more caudal and medial part. Consequently, no neurones containing both neuropeptide Y and tyrosine hydroxylase were apparent up to the day of birth. Finally, in the locus coeruleus, tyrosine hydroxylase-immunoreactive neurones were also demonstrable at day 13 of gestation. In this case, however, no neuropeptide Y-immunoreactive somata could be seen in the nucleus until day 21.

The present study indicates that the existence of neuropeptide Y and tyrosine hydroxylase in co-containing neurones is not inextricably linked, and suggests that the factors controlling the synthesis of these two substances are not identical.     

关于日本扁柏人工林光能利用效率的研究（I）——群落生产结构与现存量、生长量和光能利用效率

本研究用相关生长法与分层割取法相结合测定的21年生日本扁柏人工林的现存量为160t／ha,生长量为16.5t／ha·a,净光能利用率为0.64%。在日本福岡地区这样的光能利用率不算高。根据各层的干物质重与D2·H的相关生长关系计算了各部分的垂直分布。以太阳能多点测定计测算了林冠上部和林冠内各层太阳辐射量的季节变化。绘制了生产结构图。通过对林冠各部位叶的测定,证明了充分郁闭的日本扁柏林同日本柳杉林一样,可以区分为圆锥形、圆柱形树冠。其叶面积数和叶量密度分别为：35±5cm2／g,495g／m3;45±5cm2／g,222g／m3。以材积解析和群落生产结构、光能分布为依据论述了生产结构对光能利用的限制。     

Region- and cell type-selective expression of the evolutionarily conserved Nolz-1/zfp503 gene in the developing mouse hindbrain

Nolz-1/Zfp503, a zinc finger-containing gene, is a mammalian member of the SP1-related nocA/elb/tlp-1 gene family. Previous studies have shown that Nolz-1 homologs are important for patterning the rhombomeres in zebrafish hindbrain. We therefore studied the expression pattern of Nolz-1 in the developing mouse hindbrain. Nolz-1 mRNA expression was detected in the prospective rhombomere 3, 5 and caudal regions as early as E8.75. After E11.5, Nolz-1-positive cells were organized as distinct cell clusters, and they were largely non-overlapped with either Pax2-positive or Phox2b-positive domains. Most interestingly, we found that Nolz-1 was specifically expressed by Phox2b-negative/Isl1/2-positive somatic motor neurons, but not by Phox2b-positive/Isl1/2-positive branchial and visceral motor neurons, suggesting that Nolz-1 may regulate development of somatic motor neurons in the hindbrain. In addition to be expressed in differentiating post-mitotic neurons, Nolz-1 was also expressed by progenitor cells in the ventricular zone located in the dorsal part of aqueduct and the alar plates of hindbrain, which suggests a regulatory role of Nolz-1 in the germinal zone. Taken together, based on its domain- and cell type-selective pattern, Nolz-1 may involve in regulation of various developmental processes, including regional patterning and cell-type specification and differentiation in the developing mouse hindbrain.     

Artificial dispersal of the lichen Crocodia aurata (Lobariaceae) using asexual propagules and gel‐filled gauze packets

Lichens are an essential component of ecosystem processes. Many lichen species are habitat specialists that rely on specific tree substrates and moisture regimes of old‐growth forests and are not found in early‐successional forests, small isolated forest fragments or restoration projects, making them useful bioindicators of forest health. Pressures from habitat loss and fragmentation have affected the viability and survival of lichen species that are often limited by low dispersal ability. As a rule, lichens are rarely included in ecological restoration programmes despite translocation methods being available. We trialled a combination of two of the more successful methods (gauze packets and use of adhesive gels). Three different gelling agents were used in attempt to immobilize living soredia of the lichen Crocodia aurata on trunks of Ti Kouka/Cabbage Tree (Cordyline australis) for long enough for them to develop into thallus lobes. The effectiveness of these methods was tracked over a 2‐year period. Our results show that all three gel treatments were effective at immobilizing soredia in the gauze packets. Lobe formation occurred in all three gel treatments after 36 weeks, with methyl cellulose the most successful with six packets producing lobes, five Ac‐Di‐Sol^® and two agarose packets also producing lobes, out of a possible total 100 gauze packets. Lobe formation was slightly more successful on the south sides of trees (seven of thirteen packets), despite initial survival and vitality of soredia being higher on the north sides of the trees.