绿茶糖苷类香气前体的GC-MS和HPLC-ESI-MS~n分析

采用高效液相色谱-电喷雾多级串联质谱法(HPLC-ESI-MSn)并结合气相色谱-质谱法(GC-MS)分析鉴定了绿茶中的糖苷类香气前体物质。茶样经甲醇提取,HPD-500大孔吸附树脂分离富集糖苷,GC-MS跟踪监测酶解后挥发性苷元,然后通过电喷雾多级质谱,根据正负离子模式下的准分子离子峰和多级质谱裂解碎片,鉴定了13种糖苷类香气前体物质,其中2种糖苷首次发现存在于绿茶中。     

有机磷农药生产废水氧化塘处理的静态和动态模拟试验

马拉硫磷、对硫磷、乐果、二甲基二硫代磷酸酯和二乙基硫代磷酸酯在藻菌共生系统中是可以被分解的,它们的半分解期分别为2、5、2、42和62天。氧化塘模拟试验结果指出,废水中TOC、COD、和有机磷的去除率分别为65.1%、68.7%和67.8%。根据试验,提出了采用多级串联氧化塘处理葛店化工厂农药废水的参数以及控制鸭儿湖地区污染的方案。       

自絮凝颗粒酵母乙醇连续发酵耦合酵母回用工艺的研究

模拟现有酒精发酵行业普遍采用的多级发酵罐串联系统,建立了一套由三级串联操作的搅拌式发酵罐和两个沉降罐组成的反应器系统,以脱胚脱皮玉米粉双酶法制备的糖化液为发酵底物,培养基初始还原糖浓度为220g/L,添加(NH4)2HPO41.5g/L和KH2PO42.5g/L,以0.057h-1的恒定稀释速率流加,将自沉降浓缩后的酵母乳先后经活化和不活化两种方式处理并循环至第一级发酵罐,系统在两种操作条件下分别达到了拟稳态。实验结果表明活化处理对改善发酵工艺技术指标方面发挥了显著的作用,发酵终点乙醇浓度达到101g/L,还原糖和残总糖分别在3.2和7.7g/L左右,发酵系统的设备生产强度指标为5.77g/(L.h),与无酵母回用的搅拌式反应器系统中自絮凝颗粒酵母乙醇发酵工艺相比,提高了70%。     

一种快速准确构建酵母单杂交系统报道子的方法

酵母单杂交系统是根据DNA结合蛋白 (即转录因子 )与DNA顺式作用元件结合调控报道基因表达的原理来克隆编码目的转录因子的基因 ,其中构建带有不同DNA顺式作用元件的报道子是该系统的重要环节。目前 ,构建酵母单杂交系统报道子主要采用随机串联的方法 ,由于串联的顺式作用元件的个数和方向不能控制 ,所以筛选报道子费时费力。尝试采用同尾酶定向克隆的方法 ,将顺式作用元件按相同方向和所需的个数串联起来 ,减少了筛选报道子所需的时间和费用。     

三江平原湿地多级沟渠系统底泥可溶性有机碳的分布特征

沿着土壤和土地利用类似的三江平原多级沟渠系统采集不同沟渠单元底泥,这些不同沟渠系统单元由湿地和农田边界排水沟渠到相当大的主干渠变化着,测定底泥中可溶有机碳（DOC）含量,研究多级沟渠系统底泥中DOC的分布特征。结果发现,随沟渠等级增加,DOC在底泥中所占的比重逐渐增加（40～60cm层增加近5倍）,DOC富集层逐渐有规律的向底泥底层转移（40～60cm）。研究表明,排水沟渠既是湿地或农田DOC的最初汇聚地,又是河道DOC的输出源;沟渠重复性的排水将湿地或农田DOC通过零星径流带入沟渠系统中,连续的沟渠径流携带着大量DOC与不同等级沟渠系统相互作用使得沟渠系统底泥中DOC呈现出一定的水平、垂直分布特征;排水沟渠排水历时、DOC流量、滞留时间、径流发生频率等都会影响多级沟渠系统底泥中DOC的分布特征;另外,沟渠系统内生长的植被也通过径流阻塞和有机质过程影响底泥中DOC的分布特征。     

UASB反应器结构与性能的研究

借助于刺激响应实验,发现本中试规模(15m³)uASB的布水系统能保证反应器起动后每3．8m²有一个布水点就可使摹质分布均匀;实验验证反应器的床部和层部可分别用两个完全混合流描述,而沉降区则近似于多级串联混合流,并且在床部存在沟流(<12．2%F)和死区(7．3％Vr),层一床之间存在着反向流(300％F),三相分离器的分离功能主要由两个循环流和沉降区的作用来完成,其内部污泥的分布与分离模型相一致,分离效率在实验的有机负荷范围内保持在80％一90％。     

多级孔金属-有机框架固定化酶的研究进展

金属-有机框架(metal-organic frameworks, MOFs)作为酶固定化的优良载体,为生物催化反应提供优越的物理和化学保护。近年来,多级孔金属-有机框架(hierarchical porous metal-organic frameworks, HP-MOFs)由于其独特的结构优势,在固定化酶方面显示出更大的潜力。到目前为止,已经开发了各类具有原生多级孔或缺陷多级孔的HP-MOFs用于酶的固定化研究,并且使得固定化酶在催化活性、稳定性和重复利用性等方面得到了显著增强。本文系统总结了HP-MOFs用于固定化酶的各种策略,介绍了HP-MOFs固定化酶(enzyme@HP-MOFs)在催化合成、生物传感、生物医药等领域的最新应用进展。最后,讨论并展望了HP-MOFs固定化酶这一领域所面临的挑战和机遇。     

HPLC-ESI-MS/MS分析姜黄中姜黄素类化合物

运用HPLC-ESI-MS/MS方法分析姜黄中姜黄素类化合物。利用高效液相色谱-电喷雾多级串联质谱(HPLC-ESI-MSn)技术,以Venusil XBP C18(2.1×150 mm,5μm,Agela Technologies)作为分离色谱柱,乙腈和水为流动相,电喷雾离子源(ESI),正负离子同时扫描。根据谱峰的保留时间和质谱一、二级离子碎片信息,结合对照品及参考文献信息,同时检测出姜黄中28种姜黄素类化合物,其中有8种化合物尚未见从姜黄中报道,并解析了它们的二级质谱断裂规律。本文为更加全面研究姜黄中姜黄素类化合物提供参考。     

基因的功能冗余

1　冗余的定义对于特定的一个系统以及该系统的各种组成部件 ,其寿命总是有限的 ,不可避免地会发生故障。为了确保系统的正常运转 ,必须为系统配制一定数量的备用零件 ,即构成所谓的冗余[3 ] 。没有冗余的系统是脆弱的 ,经不起外界随机事件的干扰 ,必须具有足够的冗余 ,它才能维持其结构和功能的正常进行。冗余 (ｒｅｄｕｎｄａｎｃｅ)这一概念来源于自动控制系统可靠性理论[1] 。系统是由不同的元件 (成分 )组成的 ,元件的组合方式与系统的可靠性关系十分密切。按元件组合方式可把系统分为串联系统和并联系统。在串联系统中 ,当系统的元件…     

棘胸蛙抗菌肽Spinosan-C的串联表达与活性检测

为克服抗菌肽易被蛋白酶降解及对宿主大肠杆菌的杀伤作用,并进一步提高大肠杆菌系统的表达能力,以棘胸蛙Paa spinosa抗菌肽Spinosan-C为研究对象,按照大肠杆菌密码子利用频率进行密码子优化,设计合成8拷贝的串联8×Spinosan-C基因,将合成的串联基因克隆到大肠杆菌表达载体p ET-28a,利用大肠杆菌感受态细胞Rosetta进行原核表达,获得高效表达的串联8×Spinosan-C重组蛋白,用甲酸专一性切割得到抗菌肽Spinosan-C单体。体外抑菌试验表明,切割后的抗菌肽Spinosan-C单体对测试菌生长具有抑制作用,为蛙类抗菌肽的规模化制备提供了参考。     

Axial dispersion in plate-pulsed columns

The residence time distribution (RTD) of the phase under continuous flow and dispersed flow was measured in a plate-pulsed column under countercurrent liquid-liquid flow, and modelled using (i) the axial dispersion model and (ii) the noninteger ideal stage cascade model. The axial mixing in the continuous phase was found to be significantly higher than in the dispersed phase. The model parameters were related to the operating conditions and system variables.     

Optimization of bioreactor using metabolic control analysis approach

A new methodology based on a metabolic control analysis (MCA) approach is developed for the optimization of continuous cascade bioreactor system. A general framework for representation of a cascade bioreactor system consisting of a large number of reactors as a single network is proposed. The kinetic and transport processes occurring in the system are represented as a reaction network with appropriate stoichiometry. Such representation of the bioreactor systems makes it amenable to the direct application of the MCA approach. The process sensitivity information is extracted using MCA methodology in the form of flux and concentration control coefficients. The process sensitivity information is shown to be a useful guide for determining the choice of decision variables for the purpose of optimization. A generalized problem of optimization of the bioreactor is formulated in which the decision variables are the operating conditions and kinetic parameters. The gradient of the objective function to be maximized with respect to all decision variables is obtained in the form of response coefficients. This gradient information can be used in any gradient-based optimization algorithm. The efficiency of the proposed technique is demonstrated with two examples taken from literature: biotransformation of crotonobetaine and alcohol fermentation in cascade bioreactor system.     

High-performance recombinant protein production with <Emphasis Type="Italic">Escherichia coli</Emphasis> in continuously operated cascades of stirred-tank reactors

The microbial expression of intracellular, recombinant proteins in continuous bioprocesses suffers from low product concentrations. Hence, a process for the intracellular production of photoactivatable mCherry with Escherichia coli in a continuously operated cascade of two stirred-tank reactors was established to separate biomass formation (first reactor) and protein expression (second reactor) spatially. Cascades of miniaturized stirred-tank reactors were implemented, which enable the 24-fold parallel characterization of cascade processes and the direct scale-up of results to the liter scale. With PAmCherry concentrations of 1.15 g L^?1 cascades of stirred-tank reactors improved the process performance significantly compared to production processes in chemostats. In addition, an optimized fed-batch process was outperformed regarding space–time yield (149 mg L^?1 h^?1). This study implicates continuous cascade processes to be a promising alternative to fed-batch processes for microbial protein production and demonstrates that miniaturized stirred-tank reactors can reduce the timeline and costs for cascade process characterization.     

A continuous multistage roller reactor for animal cell culture: 1. Patterns of growth,production and catabolism of a murine hybridoma

A Tubular Liquid Film Reactor was designed as a model system to transfer a batch culture kinetic to a continuous cascade. Cell density, product formation and substrate consumption rates were followed during fermentation at two dilution rates. In spite of the high dilution rates effective in each segment by itself high cell densities of up to 10⁷ cells/ml were achieved due to cell sedimentation. The model character of the reactor was taken to determine critical values of substrate concentrations that influence production rates and result in an adaptation of metabolism.Abbreviations TLFR tubular liquid film reactor     

Molecular origin of continuous dark noise in rod photoreceptors.

Noise in the rod photoreceptors limits the ability of the dark-adapted visual system to detect dim lights. We investigated the molecular mechanism of the continuous component of the electrical dark noise in toad rods. Membrane current was recorded from intact, isolated rods or truncated, internally dialyzed rod outer segments. The continuous noise was separated from noise due to thermal activation of rhodopsin and to transitions in the cGMP-activated channels. Selectively disabling different elements of the phototransduction cascade allowed examination of their contributions to the continuous noise. These experiments indicate that the noise is generated by spontaneous activation of cGMP phosphodiesterase (PDE) through a process that does not involve transducin. The addition of recombinant gamma, the inhibitory subunit of PDE, did not suppress the noise, indicating that endogenous gamma does not completely dissociate from the catalytic subunit of PDE during spontaneous activation. Quantitative analysis of the noise provided estimates of the rate constants for spontaneous PDE activation and deactivation and the catalytic activity of a single PDE molecule in situ.     

Mathematical modelling and process optimization of a continuous 5-stage bioreactor cascade for production of poly[-(R)-3-hydroxybutyrate] by Cupriavidus necator

A multistage system for poly(hydroxyalkanoate) (PHA) production consisting of five continuous stirred tank reactors in series (5-CSTR) with Cupriavidus necator DSM 545 as production strain was modelled using formal kinetic relations. Partially growth-associated production of PHA under nitrogen limited growth was chosen as modelling strategy, thus the Luedeking-Piret’s model of partial growth-associated product synthesis was applied as working hypothesis. Specific growth rate relations adjusted for double substrate (C and N source) limited growth according to Megee et al. and Mankad-Bungay relation were tested. The first stage of the reactor cascade was modelled according to the principle of nutrient balanced continuous biomass production system, the second one as two substrate controlled process, while the three subsequent reactors were adjusted to produce PHB under continuous C source fed and nitrogen deficiency. Simulated results of production obtained by the applied mathematical models and computational optimization indicate that PHB productivity of the whole system could be significantly increased (from experimentally achieved 2.14 g L^?1 h^?1 to simulated 9.95 g L^?1 h^?1) if certain experimental conditions would have been applied (overall dilution rate, C and N source feed concentration). Additionally, supplemental feeding strategy for switching from batch to continuous mode of cultivation was proposed to avoid substrate inhibition.     

A continuous process for the production of baculovirus using insect-cell cultures

Summary A continuous culture of insect cells (Spodoptera frugiperda) was used for continuous production of baculovirus (nuclear polyhedrosis virus fromAutographa californica). The system consisted of a cascade of two continuous stirred tank reactors (CSTRs). In CSTR I the insect cells were grown in suspension. This suspension was fed continuously to CSTR II where the virus infection occurred. For a period of about 25 days the average volumetric productivity was about 10⁷ polyhedra (virus particles occluded in protein capsules) and 10⁸ infectious NOVs (non-occluded virus particles) per cm³ effluent. This is equivalent to 25 polyhedra and 250 NOVs per infected cell, respectively. In one case, the percentage of infected cells was 65%, which is close to the theoretical value of 68%. After a run-time of 32 days a decrease of process productivity was observed, probably due to the so-called passage effect, a degeneration of the virus DNA.     

Cascade stacking and cascade electrofocusing: their interconversion and fundamental unity.

The composition of a stack [an isotachophoresis (ITP) system] containing multiple trailing buffer constituents (“cascade stack”) was computed using a modification of the program of Routs. On electrophoresis, such a buffer mixture gave rise to multiple moving boundaries in which either buffer constituents or proteins could be stacked. Buffer zones within the stack served as “spacers.” The cascade stack exhibits a pH gradient, sharp zone boundaries, and constant zone width irrespective of the duration of electrophoresis, just as in the case of a stack comprising a single leading and trailing constituent. The pH gradient, sharp zone boundaries, and the sequential order of protein zones were maintained when the cascade stack was transposed between strongly acidic and basic electrolytes. Such a transposed isotachophoretic gel functioned as an electrofocusing system, indistinguishable from electrofocusing gels made in either buffers (buffer electrofocusing, or BEF) or Ampholine (isoelectric focusing, or IF). In the converse experiment, a cascade electrofocusing gel, formed in the same buffer mixture used to form a cascade stack, was subjected to electrofocusing until the steady-state was attained and then it was transposed between the appropriate upper and lower buffers of the corresponding cascade stacking system. Such transposition gave rise to moving zones with the typically sharp boundaries of a stack, a transient state pH gradient, and an order of protein zones within the cascade stack identical to the cascade electrofocusing system. These studies indicate the essential physical-chemical identity between these two types of electrophoretic systems and indicate the need for continued development of a unified theory for isoelectric focusing and steady-state stacking (isotachophoresis).     

Steady state analysis of the enhancement in ethanol productivity of a continuous fermentation process employing a protein-phospholipid complex as a protecting agent

A continuous cascade fermentation process comprising eight tanks in series, employing a protein-phopholipid complex as a protective agent (PA) was performed for ethanol production from glucose. An increase of 58.4% in fermenter productivity was obtained due to the addition of PA. A kinetic model including product and substrate inhibition effects is proposed. Parameters appearing in the kinetic model were estimated by using the method of least squares. It is found that the product inhibition effect dominates over the substrate inhibition effect for the range of concentrations studied in our fermentation system. Upon addition of PA, both inhibitory effects are reduced to as little as about one quarter of that without PA. It was also found that the use of PA primarily protected the cells against ethanol inhibition rather than substrate inhibition. A steady state criterion is also discussed.