应用免疫印迹法分析丙型肝炎相关性抗体

应用两种免疫印迹法对二代ELISA测试的69份血清进行了追加试验。结果表明:8份ELISA阴性血清免疫印迹均阴性,61份ELISA阳性血清有1份免疫印迹阴性。ELISA假阳性率为1.67％。在60份免疫印迹阳性血清中,抗结构区(capsid),非结构区NS_3和NS_4抗体的阳性率分别为83％、93.3％和70％。其中丙型肝炎相关性抗体的分布,以抗capsid、NS_3、NS_4抗体都阳性为主(58.7％)。在输血后急性肝炎和肝细胞癌患者中。丙型肝炎相关性抗体的分布没有统计学上的差异。     

提高卷烟吸味降低焦油释放量的应用研究

为降焦减害提高卷烟吸味,利用现代生物技术,在卷烟制丝过程中加入一定量的MJQ酶制剂,通过正交试验选择最优条件,即温度为100 ℃、贮存时间为24 h、浓度为1 %.经后续对比试验及焦油测试与评吸,可以验证在卷烟制丝过程中喷洒MJQ酶制剂,对卷烟的焦油量有明显的降低,卷烟吸味得到了明显的改善.     

A lysine to arginine substitution at position 146 of rabbit aldolase A changes the rate-determining step to Schiff base formation

Lys146 of rabbit aldolase A [D-fructose-1,6-bis(phosphate):D-glyceraldehyde-3-phosphate lyase, EC 4.1.2.13 [EC] ] was changedto arginine by site-directed mutagenesis. The k_cat of the resultingmutant protein, K146R, was 500 times slower than wild-type insteady-state kinetic assays for both cleavage and condensationof fructose-1,6-bis(phosphate), while the Km for this substratewas unchanged. Analysis of the rate of formation of catalyticintermediates showed K146R was significantly different fromthe wild-type enzyme and other enzymes mutated at this site.Single-turnover experiments using acid precipitation to trapthe Schiff base intermediate on the wild-type enzyme failedto show a build-up of this intermediate on K146R. However, K146Rretained the ability to form the Schiff base intermediate asshown by the significant amounts of Schiff base intermediatetrapped with NaBH₄. In the single-turnover experiments it appearedthat the Schiff base intermediate was converted to productsmore rapidly than it was produced. This suggested a maximalrate of Schiff base formation of 0.022 s^–1, which wasclose to the value of k_cat for this enzyme. This observationis strikingly different from the wild-type enzyme in which Schiffbase formation is >100 times faster than k_cat. For K146Rit appears that steps up to and including Schiff base formationare rate limiting for the catalytic reaction. The carbanionintermediate derived from either substrate or product, and theequilibrium concentrations of covalent enzyme-substrate intermediates,were much lower on K146R than on the wild-type enzyme. The greaterbulk of the guanidino moiety may destabilize the covalent enzyme-substrateintermediates, thereby slowing the rate of Schiff base formationsuch that it becomes rate limiting. The K146R mutant enzymeis significantly more active than other enzymes mutated at thissite, perhaps because it maintains a positively charged groupat an essential position in the active site or perhaps the Argfunctionally substitutes as a general acid/base catalyst inboth Schiff base formation and in subsequent abstraction ofthe C4-hydroxyl proton.     

“绿色”溶剂离子液的研究进展

近年来，离子液由于其独特的优点, 在生物催化及化学合成领域越来越受到青睐。由于缺乏蒸气压，离子液作为绿色溶剂有很大潜能。此外，离子液不像极性有机溶剂那样会使酶失活，因而简化了类似含有糖类这样极性底物的反应。在离子液中，生物催化反应表现出较高的选择性、较快的反应速率和较高的酶稳定性。但这些溶剂还存在着离子液的纯化、控制水活性和pH、高粘度以及产品分离等问题。     

温敏性微凝胶对蛋白质和酶的吸附性能

用微乳液聚合方法以N-异丙基丙烯酰胺为单体合成了温敏性微凝胶聚N-异丙基丙烯酰胺(PNIPAM),研究了其对两种蛋白质和两种酶的吸附性能,测定了吸附等温线和温度对吸附量的影响。结果表明,微凝胶在低临界溶解温度(LCST)附近吸附蛋白质和酶的量有一突跃,例如在LCST前后,1 g纳米颗粒吸附的酪蛋白的质量分别为225 mg和415 mg;吸附的枯草杆菌蛋白酶的质量分别为12 000U/mg和27 500 U/mg。蛋白质和酶是通过物理吸附作用结合到PNIPAM微凝胶上,可以用调节温度的方法,来控制温敏微凝胶对蛋白质和酶的吸附与脱附。     

Theoretical examination of the mechanism of aldose-ketose isomerization

Two mechanisms for an aldose–ketose isomerization havebeen examined using high level ab initio and semiempirical molecularorbital methods. The proton transfer pathway via an enediolintermediate is shown to be favored in the absence of a metalion, while the hydride transfer pathway becomes favored in thepresence of a metal ion. Our calculations explain why the protontransfer pathway is operative in most aldose–ketose isomerizationreactions. These calculations also provide further support forthe previously proposed metal ion-mediated hydride transfermechanism for xylose isomerase.     

内部连接的球形多孔羟基磷灰石支架的制备

采用高分子模板,通过注浆成型制备了球形孔结构的羟基磷灰石(hroxyapatite,HA)支架,研究了支架的微观结构、压缩强度和相容性.结果表明:改变工艺参数可以控制高分子模板中球体间胶黏面的大小,控制所制备的HA多孔体孔内连接的尺寸.制备的支架具有开口、均匀而内部连通的多孔结构,当孔径为200μm、孔隙率为65%时,压缩强度可达8MPa左右.体外生物模拟实验表明:多孔结构HA支架有很好的相容性.随多孔球体孔隙率和尺寸的增加,支架的细胞吸收率逐步增加.     

Flaking and extrusion as mechanical treatments for enzyme-assisted aqueous extraction of oil from soybeans

Flaking and extruding dehulled soybeans were evaluated as a means of enhancing oil extraction efficiency during enzyme-assisted aqueous processing of soybeans. Cellulase, protease, and their combination were evaluated for effectiveness in achieving high oil extraction recovery from extruded flakes. Aqueous extraction of extruded full-fat soy flakes gave 68% recovery of the total available oil without using enzymes. A 0.5% wt/wt protease treatment after flaking and extruding dehulled soybeans increased oil extraction recovery to 88% of the total available oil. Flaking and extruding enhanced protease hydrolysis of proteins freeing more oil. Treating extruded flakes with cellulase, however, did not enhance oil extraction either alone or in combination with protease. Discrepancies in oil extraction recoveries were encountered when merely considering crude free fat because some oil became bound to denatured protein during extrusion and/or sample drying. Bound fat was unavailable for determination by using the hexane extraction method, but was accounted for by using the acid hydrolysis method for total oil determination. Oil extraction recovery from extruded soybean flakes was affected by oil determination methods, which was not the case for unextruded full-fat soy flour.     

影响酶洗涤效果因素的探讨

介绍了加酶洗涤剂的发展以及酶在洗涤剂中的应用。探讨了加酶洗涤剂中表面活性剂及其浓度、洗涤温度和洗涤时间对洗涤效果的影响，pH值对酶活性的影响。对影响酶洗涤效果的各种因素进行了阐述。并论述了酶在洗涤剂中的其他应用。     

Properties of cuticular oxidases used for sex pheromone biosynthesis byHeliothis zea

Biosynthesis of the aldehydic sex pheromone components released by females ofHeliothis zea was found to be catalyzed by primary alcohol oxidases residing in the cuticle that covers the glands. Activity, as indicated by conversion of primary alcohol to aldehyde, was as high in cell-free cuticle as it was in intact pheromone glands. Studies indicated that some activity was associated with the surface of the epicuticle and could be removed, into buffer, by sonication. However, the majority of activity lies within the inner epicuticle and exo- and endocuticular layers. The oxidase was not functional in pharate pupae that did not have mature adult cuticle but became functional just prior to adult emergence. The enzyme in individual glands was saturated at alcohol concentrations above 100 n. moles. Nonionic detergents did not affect the activity of the oxidase in the cuticle but treatment with either 7 M urea or 1% SDS resulted in total loss of activity. Studies on the effect of pH indicated an optimum at 6.4; however, activity was high throughout the range of 5–9. The oxidase was functional in both dichloromethane and hexane, suggesting that this enzyme system may have applications for organic synthesis of aldehydes.