枯草杆菌6-磷酸葡萄糖酸脱氢酶的分离纯化及部分性质研究

采用DEAE—Sephaurose离子交换层析，Blue-Seplaarose CL-6B特异结合层析和Sephadex G-200凝胶过滤技术，对枯草芽孢杆菌中的6-磷酸葡萄糖酸脱氢酶进行了分离纯化，纯化倍数为112．3倍，比活为1．46U／mg，回收率为8．2％，纯化酶液经聚丙烯酰胺凝胶电泳检验为单一带，测得全酶相对分子质量为107kD，具有两个分子量相同的亚基，亚基相对分子质量为51kD，最适pH值为8．0，最适温度为30℃，对热不稳定，以6-磷酸葡萄糖酸和NADP^ 为底物，其Km值分别为Km1(NADP^ )=19．8μmol／L和Km2(6-GPA)=22．6μmol／L，在5mmol／L～50mmol／L浓度范围内，Mg^ ，Ca^2 和Mn^2 对酶有激活作用，Fe^3 和Cu^2 对酶有抑制作用，K^ ，Na^ ，Cl^-，NO^-3，SO^-4对酶几乎没有影响，用PMSF，TNBS，NBS，DTT和BrAc对酶进行修饰，实验结果表明，丝氨酸、苏氨酸、赖氨酸和组氨酸残基可能是该酶活性中心的功能基团。     

不对称还原苯乙酮酸甲酯的生物转化菌种选育及条件优化

选用酵母菌、乳酸菌、粪链球菌、假丝酵母等4个种属共15株菌株,研究其对苯乙酮酸甲酯的不对称还原催化特性.以具有R(-)-扁桃酸甲酯较高转化活力的菌株S.cNo.1、S.cNo.3、S.cNo.9作为出发菌株,采用紫外与微波复合诱变的方法,筛选获得S.c3.5.16突变株.考察培养温度、pH值对该突变株转化活力的影响,结果表明,S.c3.5.16菌株的R(-)-扁桃酸甲酯生物转化最适反应条件为培养基pH6.5、温度38℃,苯乙酮酸甲酯转化为扁桃酸甲酯的得率达到99.3%,R(-)-扁桃酸的光学纯度达到95.5%e.e..     

产朊圆酵母尿酸酶的分离纯化及其部分性质研究

产朊圆酵母Y0401经尿酸诱导培养,菌体采用超声波处理,依次经过DEAE-Sepharose离子交换层析、Phenyl Sepharose CL-4B疏水层析和Xanthine-agarose亲和层析,尿酸酶比活力达到18U/mg,纯化倍数为409倍,回收率为19.2%.纯化后的酶经还原、非还原SDS-PAGE分析为单一蛋白染色带,HPLC显示纯度为98%以上.用HPLC和Superdex 200分子筛层析两种方法测得该酶的相对分子质量为127kDa,在还原条件下进行SDS-PAGE,测得表观分子质量为33kDa,综上结果推测该酶是由分子量相同的4个亚基组成的四聚体蛋白.等电聚焦显示其等电点在6.8~7.5之间.该酶具有较强的酸碱稳定性和热稳定性.最适pH为8.5,最适温度为40.0℃.在最适pH和温度条件下测得该酶Km值为3.34×10-5mol/L.对某些金属离子和有机物对酶活性的影响也进行了研究.     

青枯雷尔氏菌龙胆酸1,2-双加氧酶基因的克隆、表达和酶性质的研究

本研究克隆、表达了青枯雷尔氏菌GMI1000菌株中编码龙胆酸1,2-双加氧酶的基因, 并通过亲和层析对该酶进行了纯化, SDS-PAGE结果表明该酶亚基约为38kDa.该酶的最适反应温度和最适pH分别为30℃和7.5.该酶的Km为56μmol/L,pI为4.6～4.8.纯化后的龙胆酸1,2-双加氧高度不稳定:4℃下放置72h即失去85%的酶活. 甘油和β-巯基乙醇可稳定该酶酶活,在添加10%(体积比)甘油至酶液(50mmol/L, pH 7.3的磷酸盐缓冲液保存)后,-20℃保藏2周后均能检出明显的酶活.0.1～1mmol/L Fe2 可以激活或者稳定该酶的酶活.Na ,K ,Mg2 和Ca2 (分别为1～2mmol/L)对该酶的酶活无明显的影响.Mn2 ,Zn2 和Fe3 的添加量超过2mmol/L时,酶活急剧下降.1mmol/L的Cu2 即使该酶失去酶活.     

克雷伯杆菌1,3-丙二醇氧化还原酶的分离纯化及其酶学性质

在有氧条件下,利用DEAE Sepharose Fast Flow弱阴离子离子交换层析和Blue Sepharose CL-6B亲和层析,同时分离并提纯克雷伯杆菌胞内的1,3-丙二醇氧化还原酶和甘油脱氢酶.研究表明,1,3-丙二醇氧化还原酶的纯化倍数为35.86倍,回收率为5.17%,该酶最适表观反应温度为57 ℃,最适反应pH值为9.5.在30 ℃及pH=8.0～10.0时,该酶具有良好的稳定性.在45 ℃和pH=9.5条件下,该酶以1,3-丙二醇和NAD+为底物,其米氏常数Km分别为15.8,0.2 mmol·L-1.1,3-丙二醇氧化还原酶对生理反应底物3-羟基丙醛活性最大,对其他醇类也有氧化能力.Mn2+对酶有显著激活作用,巯基保护剂能明显提高酶的活力.     

芦荟叶皮β-葡萄糖苷酶的分离纯化及酶学性质

库拉索芦荟叶皮经匀浆、柠檬酸-柠檬酸钠缓冲液抽提,硫酸铵分级沉淀,DEAE-Sepharose fast flow层析和Superdex-200prep grade层析,获得电泳纯的β-葡萄糖苷酶(β-Glucosidase,BGL).纯化结果是β-葡萄糖苷酶比活力为98.48U/mg,纯化倍数为328.27倍,酶活性回收率为9.87%,全酶分子质量约为69.3kD,亚基分子质量约为69.7kD.酶学性质研究表明:β-葡萄糖苷酶的最适反应温度和最适反应pH值分别为40℃和5.0;在20~30℃及pH4.0~8.0范围内稳定性较好;最适条件下,以pNPG为底物的K_m值为2.21mmol/L,V_(max)为1.381μmol/(min·L).乙醇,抗坏血酸,Mn~(2+),K~+对该酶活性具有激活作用;SDS,Cu~(2+)对该酶活性具有强烈抑制作用;异丙醇,EDTA,尿素,Mg~(2+),Li+对该酶活性影响较小;甲醇对该酶有双重作用.     

微生物转化苯乙酮酸合成R-(-)- 扁桃酸

考察了酵母菌株FD11b不对称还原苯乙酮酸生产R-(-)-扁桃酸的重复使用性能.菌株FD11b在持续6批次(14 d)的还原反应中,一直保持着较高的催化还原活性,且目标产物R-(-)-扁桃酸的对映体过量值(e.e.值)均高于96.5%.在30 L发酵罐上进行Fed-Batch放大试验,当菌体浓度(细胞干重,以下同)为12.5 g/L,底物苯乙酮酸浓度控制在20~30 mmol/L时,还原反应49 h,产物浓度可达125 mmol/L,产物得率为96.1%,e.e.值为98.5%.     

农药靶标乙酰胆碱酯酶的分离纯化及性质研究

采用了Sephadex G-25层析,DEAE-Sepharose Fast Flow离子交换层析和Sephacryl S-200凝胶过滤层析,对家蚕头部粗酶匀浆液中的乙酰胆碱酯酶进行纯化,得到的样品经聚丙烯酰胺凝胶电泳（PAGE）和SDS-PAGE检测均为一条带,用SDS-PAGE法测得其分子量为77.8 kDa.酶的最适反应温度为37 ℃,最适pH为7.5,对底物碘化硫代乙酰胆碱(ATChI)的Km值为0.392 mmol/L,最适底物浓度为1.6 mmol/L,在1.6 mmol/L底物浓度以上观察     

产朊假丝酵母(Candida utilis)尿酸酶的基本特性

产朊假丝酵母尿酸酶通过由DEAE-sepharose和Xanthine-agarose等层析纯化.纯化酶经SDS-PAGE显示单一蛋白带,HPLC显示纯度为97.5%.其相对分子质量为127 kDa,变性SDS-PAGE表明该酶的表观分子质量为33 kDa.该酶具有较强的酸碱稳定性和热稳定性,最适pH为8.5,最适温度为40.0℃.在最适条件下测得其Km为35.0 μmol/L.某些金属离子对酶活性的研究结果表明,Co3 ,Pb2 ,Mn2 及EDTA,NEMI对尿酸酶活性有抑制作用,Mg2 ,Triton X-100对尿酸酶活性略有激活作用.     

交联海藻酸钙凝胶固定化酵母醇脱氢酶研究

对用海藻酸钙包埋、戊二醛交联法固定酵母醇脱氧酶催化苯乙酮酸合成(R)-扁桃酸的过程进行研究,比较游离酶与固定化酶的酶学性质.实验结果表明:固定化酶的热稳定性显著提高,游离酶在70℃时酶蛋白变性失去活力,而固定化酶在65℃保温1 h的能保持64%的酶活力,在70℃时酶活力仍町保留48.6%;固定化酶的最适温度由30℃升至40℃,最适反应pH值由6.8下降为5.8;固定化酶保留了62.72%的游离酶活性, 固定化酶的表观米氏常数和最大反应速率分别为37.33 mmol/L和358.42 nmol/min.该固定化酶具有良好的储存稳定性和操作稳定性.     

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.     

Lower Paleozoic oil relationships within Williston Basin, Canada

The Williston Basin is a significant petroleum province, containing oil production zones that include the Middle Cambrian to Lower Ordovician, Upper Ordovician, Middle Devonian, Upper Devonian and Mississippian and within the Jurassic and Cretaceous. The oils of the Williston Basin exhibit a wide range of geochemical characteristics defined as "oil families", although the geochemical signature of the Cambrian Deadwood Formation and Lower Ordovician Winnipeg reservoired oils does not match any "oil family". Despite their close stratigraphic proximity, it is evident that the oils of the Lower Palaeozoic within the Williston Basin are distinct. This suggests the presence of a new "oil family" within the Williston Basin. Diagnostic geochemical signatures occur in the gasoline range chromatograms, within saturate fraction gas chromatograms and biomarker fingerprints. However, some of the established criteria and cross-plots that are currently used to segregate oils into distinct genetic families within the basin do not always meet with success, particularly when applied to the Lower Palaeozoic oils of the Deadwood and Winnipeg Formation.     

An Analysis of Catherine＇s Character——Wuthering Heights

Wuthering Heights, Emily Bronte＇s only novel, was published in December of 1847 under the pseudonym Ellis Bell. The book did not gain immediate success, but it is now thought one of the finest novels in the English language. Catherine is the key character of this masterpiece, because everybody and everything center on her though she had a short life. We can understand this masterpiece better if we know Catherine well.     

Characterization of Ordovician carbonate reservoirs, southeastern Saskatchewan, Canada

The discovery of the prolific Ordovician Red River reservoirs in 1995 in southeastern Saskatchewan was the catalyst for extensive exploration activity which resulted in the discovery of more than 15 new Red River pools. The best yields of Red River production to date have been from dolomite reservoirs. Understanding the processes of dolomitization is, therefore, crucial for the prediction of the connectivity, spatial distribution and heterogeneity of dolomite reservoirs.The Red River reservoirs in the Midale area consist of 3～4 thin dolomitized zones, with a total thickness of about 20 m, which occur at the top of the Yeoman Formation. Two types of replacement dolomite were recognized in the Red River reservoir: dolomitized burrow infills and dolomitized host matrix. The spatial distribution of dolomite suggests that burrowing organisms played an important role in facilitating the fluid flow in the backfilled sediments. This resulted in penecontemporaneous dolomitization of burrow infills by normal seawater. The dolomite in the host matrix is interpreted as having occurred at shallow burial by evaporitic seawater during precipitation of Lake Almar anhydrite that immediately overlies the Yeoman Formation. However, the low δ18O values of dolomited burrow infills (-5.9‰～ -7.8‰, PDB) and matrix dolomites (-6.6‰～ -8.1‰, avg. -7.4‰ PDB) compared to the estimated values for the late Ordovician marine dolomite could be attributed to modification and alteration of dolomite at higher temperatures during deeper burial, which could also be responsible for its 87Sr/86Sr ratios (0.7084～0.7088) that are higher than suggested for the late Ordovician seawaters (0.7078～0.7080). The trace amounts of saddle dolomite cement in the Red River carbonates are probably related to "cannibalization" of earlier replacement dolomite during the chemical compaction.     

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.     

Design and analysis of generic LBS application developing platform

Location based services is promising due to its novel working style and contents.A software platform is proposed to provide application programs of typical location based services and support new applications developing efficiently. The analysis shows that this scheme is easy implemented, low cost and adapt to all kinds of mobile nework system.     

橡胶颗粒沥青混合料低温抗裂性试验研究

以AC-13级配为基础,将橡胶颗粒代替部分集料掺入混合料中,以低温弯曲试验为评价方法对不同橡胶颗粒掺量下沥青混合料的低温抗裂性进行研究,并引入应变能密度值对混合料的低温抗裂性进行综合评价.试验结果表明:橡胶颗粒沥青混合料试件的破坏微应变均超过2 300,满足冬寒区的技术指标;无论是否掺加橡胶颗粒,随着温度的下降,沥青混合料破坏时的最大弯拉强度增大,弯拉应变降低,劲度模量增大;弯曲应变能密度在胶粒掺量为1%左右时具有较大的弯曲应变能密度值,此时橡胶颗粒沥青混合料具有较好的低温抗裂性.     

A computer generator for randomly layered structures

AcomputergeneratorforrandomlylayeredstructuresYUJia shun1,2,HEZhen hua2(1.TheInstituteofGeologicalandNuclearSciences,NewZealand;2.StateKeyLaboratoryofOilandGasReservoirGeologyandExploitation,ChengduUniversityofTechnology,China)Abstract:Analgorithmisintrod…     

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。     

Quantitative trait loci （QTLs） for quality traits related to protein and starch in wheat

Quality traits in wheat （Triticum aestirum L.） were studied by quantitative trait locus （QTL） analysis in a recombinant inbred line （RIL） population, a set of 131 lines derived from Chuan 35050 × Shannong 483 cross （ChSh）. Grains from RILs were assayed for 21 quality traits related to protein and starch. A total of 35 putative QTLs for 19 traits with a single QTL explaining 7.99-40.52% of phenotypic variations were detected on 10 chromosomes, 1D, 2A, 2D, 3B, 3D, 5A, 6A, 6B, 6D, and 7B. The additive effects of 30 QTLs were positive, contributed by Chuan 35050, the remaining 5 QTLs were negative with the additive effect contributed by Shannong 483. For protein traits, 15 QTLs were obtained and most of them were located on chromosomes 1 D, 3B and 6D, while 20 QTLs for starch traits were detected and most of them were located on chromosomes 3D, 6B and 7B. Only 7 QTLs for protein and starch traits were co-located in three regions on chromosomes 1D, 2A and 2D. These protein and starch trait QTLs showed a distinct distribution pattern in certain regions and chromosomes. Twenty-two QTLs were clustered in 6 regions of 5 chromosomes. Two QTL clusters for protein traits were located on chromosomes 1D and 3B, respectively, three clusters for starch traits on chromosomes 3D, 6B and 7B, and one cluster including protein and starch traits on chromosome 1D.