克鲁维酵母(K.hubeiensis)产菊粉酶的发酵参数研究

菊粉酶(inulinase)是一种降解菊糖B-2,1-D-果聚糖果糖苷键生成果糖或低聚果糖的水解酶.本研究在筛选菊粉酶高产菌株基础上,对产酶能力较强的l株湖北克鲁维酵母(Kluyvreromyces hubeiensis)的发酵产酶参数进行了优化;结果表明,该菌株在菊糖3%,蛋白胨3%,pH 5.0,30～37℃摇床培...     

克鲁维酵母中外切菊粉酶基因在毕赤酵母中的表达研究

用PCR的方法从克鲁维酵母(Kluyveromyces marxianus)ATCC12424中克隆出外切菊粉酶基因.将该基因克隆到毕赤酵母表达载体pHBM905C、pHBM906上,构建了外切菊粉酶毕赤酵母表达载体pHBM1200、pHBM1201.将两者转化毕赤酵母GS115,得到重组菌株GS115(pHBM1200)、GS115(pHBM1201),将这两种重组菌株进行摇瓶发酵,测得带α-信号肽的菌株GS115(pHBM1200)表达的外切菊粉酶最高酶活为89.43 U/mL,带自身信号肽的菌株GS115(pHBM1201)表达的外切菊粉酶最高酶活为14.828 U/mL.SDS-PAGE电泳表明,外切菊粉酶的表观分子量为90 kD.将外切菊粉酶用去糖基化酶endoH处理后,SDS-PAGE电泳表明,分子量为60 kD,和预计的分子量一致.     

利用菊粉产山梨醇酵母融合菌株的构建

介绍高产菊粉酶的克鲁维酵母(Kluyveromycessp.)Y85菌株与产山梨醇的酿酒酵母(Saccharomycescerevisiae)E15菌株,通过属间原生质体融合构建直接利用菊粉产山梨醇的酵母融合菌株.结果认为,以0.1%EDTA-巯基乙醇为脱壁预处理剂,蜗牛酶浓度2%(W/V),酶解温度30℃,E15和Y85两菌株细胞酶解时间分别为40min和30min,原生质体形成率和再生率分别达90%和25%以上.融合的适宜条件为添加30%(W/V)PEG和10mmol/LCaCl2,融合时间为30min,融合率可达2.64×10-6以上.利用两亲株自然性状差异检出融合菌株,并经DNA含量、菊粉酶活性、细胞形态与大小、菌落形态等测定加以确证.其中F27融合菌株在适宜条件下,山梨醇摇瓶发酵产量可达4.87g/100mL,并且具有遗传稳定性能.     

克鲁维酵母y—85菊粉酶水解菊粉的研究及其中试

克鲁维酵母（Ｋｌｕｖｅｒｏｍｙｃｅｓｓｐ．）Ｙ－８５在菊粉提取液培养基中生长产菊粉酶，其细胞和胞外和炙粉酶活力分别约占菊粉酶总活力的７６％和２４％，试验中以酶发液酶液，该酶的Ｓ／Ｉ值为１５．２酶液在５０，５５与６０℃下保温１ｈ，活力分别残留１００％，９６％和６５％，在８℃下放置７ｄ１４ｄ酶活力分别残留９８％和９６％，在ｐＨ３．６～７．０内，酶活性十发稳定，５ｍｍｏｌＬ－半胱氨酸和Ｆｅ＾＋２分别可使     

内切菊粉酶基因在毕赤酵母中的高水平表达研究

低聚果糖(Fructooligosaccharides,FOS)具有改善胃肠道环境、刺激和加强机体免疫反应、抑制肠道特定肿瘤的发生等生物活性.内切菊粉酶能够水解菊粉获得低聚果糖.为了实现内切菊粉酶的高表达,从无花果曲霉(Aspergillus ficuum ATCC16882)中克隆了内切菊粉酶编码基因INU2,并实现了在毕赤氏酵母中的重组分泌表达,摇瓶表达的酶活力为570.4U/mL.去除内切菊粉酶的内源性信号肽序列后,其重组表达水平显著提高,摇瓶表达的酶活力为1013.8U/mL,提高幅度77.7%.TLC分析表明:毕赤酵母重组表达的内切菊粉酶(不含内源性信号肽序列)能将2%长链菊粉水解为2～3个聚合度的低聚果糖.     

克鲁维酵母Y－85菊粉酶的研究

菊粉酶产生菌株─—克鲁维酵母Ｙ－８５在１５Ｌ罐中进行产酶发酵，培养２４ｈ，发酵液中菊粉酶的总活力可达１０３２ｕ／ｍｌ，其中胞壁酶活７６８ｕ／ｍｌ，占总酶活的７４％。用含半胱氨酸的缓冲液提取胞壁酶，酶的回收率达８８％，比活力可达１００８ｕ／ｍｇ．酶作用于菊糖的最适ｐＨ为４．５，最适温度为５０℃，在５０℃以下和ｐＨ４～８稳定，金属离子Ａｇ￣＋、Ｈｇ￣（２＋）对酶有强烈的抑制作用，ＰＣＭＢ对酶也有明显的抑制作用。该酶可水解菊糖，棉子糖和蔗糖等多种底物，能快速将菊芋抽提液中的菊糖水解成果糖，适合用于以菊芋为原料的高果糖浆生产。     

菊粉内切酶在毕赤酵母中的表达及酶学性质测定

为了实现菊粉内切酶在毕赤酵母(Pichia pastoris)中的高活性表达并对表达的菊粉内切酶酶学性质、水解菊粉的产物进行分析。通过下载无花果曲霉菊粉内切酶基因的编码序列,做了密码子优化后进行全基因合成,然后在毕赤酵母GS115中表达。对表达的菊粉内切酶的酶活、最适反应温度、最适pH值、热稳定性进行了分析,最后对酶水解菊粉的产物进行了高效液相色谱(HPLC)分析。摇瓶表达的酶活力为420 U/mL,酶的最适反应温度为55℃,最适反应pH为6.0。HPLC分析表明:表达的菊粉内切酶酶解底物菊粉的主要产物为低聚果糖,聚合度为3～5之间。构建的工程酵母可以高效表达菊粉内切酶,可以用于生产低聚果糖。     

克鲁维酵母y－85菊粉酶水解菊粉的研究及其中试

克鲁维酵母（Ｋｌｕｖｅｒｏｍｙｃｅｓｓｐ．）Ｙ－８５在菊粉提取液培养基中生长产菊粉酶，其细胞和胞外的菊粉酶活力分别约占菊粉酶总活力的７６％和２４％．试验中以酶发酵液作酶液．该酶的Ｓ／Ｉ值为１５．２．酶液在５０，５５和６０℃下保温１ｈ，活力分别残留１００％，９６％和６５％；在８℃下放置７ｄ，１４ｄ，酶活力分别残留９８％和９６％．在ｐＨ３．６～７．０内，酶活性十分稳定．５ｍｍｏｌＬ－半胱氨酸和Ｆｅ＋２分别可使酶活力提高１０．４％和４１．２％．ｙ－８５菊粉酶水解菊粉的适宜条件是：底物为总糖浓度１０％～１５％菊粉提取液，ｐＨ５．０，温度为５５℃，酶用量为底物每克糖加酶８００ｕ（以蔗糖为底物测酶活力）或２６．３ｕ（以菊糖为底物测酶活力）、搅拌酶解６ｈ．在上述条件上，底物降解率最高可达９８．５％．酶解液中，果糖占总还原糖量的８５％．用１１００Ｌ罐进行酶解中试，５批试验，底物降解率平均达９４．２％．     

激光复合诱变黑曲霉原生质体选育热稳定菊粉酶高产菌

以菊粉酶产生菌黑曲霉ＡＳ－４原生质体为对象,经激光和亚硝基胍复合诱变,从大量突变株中最终选育出高产热稳定的菊粉酶产生菌ＡＬ－１５４．在以菊芋提取液为碳源、玉米浆加０．３％酵母膏为氮源的基本培养基中,菊粉酶活力达１４６．３ｕ／ｍＬ,比出发株提高近３倍．其产酶最适营养条件为：起始ｐＨ４．５～５．０,２８～３０℃,３６ｈ时产酶量达最高．酶作用最适条件为ｐＨ５．０,６０℃;该酶遗传性能稳定,热稳定性强,６０℃保温处理２ｈ,其酶活性不变,６５℃保温３０ｍｉｎ,仍保留９８％的初始酶活     

海洋细菌NTa发酵产琼胶酶条件的初步优化

利用选择性培养基从厦门红树林泥土样品中分离得到一株具有较高琼胶酶活力的海洋细菌NTa,通过16S rRNA分析,将该菌株归属为Stenotrophomonas sp.NTa.对该菌株发酵产琼胶酶的条件进行初步优化,结果表明:菌株NTa产琼胶酶的最佳碳源是琼脂,氮源是酵母浸膏,NaCl的质量分数为5%,在28 ℃发酵培养40 h,发酵液的酶活力达到1.98 U/mL,比优化前提高了37.35%.     

Markedness and its Interpretation in Language Use

Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.     

Features of Women's language

Language is a means of verbal communication. People use language to communicate with each other. In the society, no two speakers are exactly alike in the way of speaking. Some differences are due to age, gender, statue and personality. Above all, gender is one of the obvious reasons. The writer of this paper tries to describe the features of women＇s language from these perspectives： pronunciation, intonation, diction, subjects, grammar and discourse. From the discussion of the features of women＇s language, more attention should be paid to language use in social context. What＇s more, the linguistic phenomena in a speaking community can be understood more thoroughly.     

低渗透非均质砂岩油藏启动压力梯度研究

理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。     

On Tragic Consciousness In The Works By Ernest Hemingway

As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.     

Achieving Unusual States of Matter Under High Pressure

正The periodicity of the elements and the non-reactivity of the inner-shell electrons are two related principles of chemistry,rooted in the atomic shell structure.Within compounds,Group I elements,for example,invariably assume the+1 oxidation state,and their chemical properties differ completely from those of the p-block elements.These general rules govern our understanding of chemical structures and reactions.Using first principles calcula-     

Exchange-Correlation and Electronic Excitation Energies from Pairing Matrix Fluctuations

We have developed an adiabatic connection to formulate the ground-state exchange-correlation energy in terms of pairing matrix linear fluctuations.This formulation of the exchange-correlation energy opens a new channel for density functional approximations based on the many-body perturbation theory.We illustrate the potential of such approaches with an approximation based on the particle-particle Random Phase Approximation(pp-RPA).This re-     

Density Functional Theory for the Response of Periodic Systems to Electric Fields Based on the Vector Potential Approach

正The electronic and nuclear(structural/vibrational)response of 1D-3D nanoscale systems to electric fields gives rise to a host of optical,mechanical,spectral,etc.properties that are of high theoretical and applied interest.Due to the computational difficulty of treating such large systems it is convenient to model them as infinite and periodic(at least,in first approximation).The fundamental theoretical/computational problem in doing so is that     

On the Thermodynamic Limit for Responses to Static Electromagnetic Fields

For molecular systems,the quantum-mechanical treatment of their responses to static electromagnetic fields usually employs a scalar-potential treatment of the electric field and a vector-potential treatment of the magnetic field.Although the potential for each field separately is associated with the choice of an(unphysical)origin,the precise choice of the origin for the electrostatic field has little consequences for the results.This is different for the     

An affordable accurate evaluation of the binding conformations predicted by molecular docking studies in bio-system

Franck-Condon factors bridge the gap between theoretical modeling and experimental observations for molecular electronic spectroscopy and electron transfer.Under the displaced harmonic oscillator approximation,multidimensional Franck-Condon factors are decomposed into a product of many one-dimensional(1D)Franck-Condon(FC)factors,and each 1D-FC factor is associated with one Huang-Rhys factor that determines the leading contribution of     

Call for Papers

<正>"The Journal of Shanghai Normal University:Mathematics"is published by Shanghai Normal University as regular issues of The Journal of Shanghai Normal University each year from 2014 in English.The editors-in-chief of the issues are professors Yuhao Cong and Maoan Han.The Journal of Shanghai Normal University was started in 1958 with