为了提高甘露聚糖酶ManA在毕赤酵母中分泌表达的酶活,选择毕赤酵母内质网未折叠蛋白反应(Unfolded protein response,UPR)激活调控因子HAC1与5种毕赤酵母蛋白折叠相关的分子伴侣ERO1、PDI、PDI1、CPR5、BiP,通过构建pPICZA-HAC1等6种胞...为了提高甘露聚糖酶ManA在毕赤酵母中分泌表达的酶活,选择毕赤酵母内质网未折叠蛋白反应(Unfolded protein response,UPR)激活调控因子HAC1与5种毕赤酵母蛋白折叠相关的分子伴侣ERO1、PDI、PDI1、CPR5、BiP,通过构建pPICZA-HAC1等6种胞内表达重组质粒,分别电转化至分泌表达ManA的毕赤酵母重组菌中胞内共表达,并分析其重组菌摇瓶发酵时ManA表达的影响。结果发现在摇瓶发酵水平,胞内共表达HAC1、ERO1、PDI的重组菌发酵上清液中的ManA酶活力分别提高了26%、15%、20%,其重组菌发酵上清液的酶活力分别达到1 014 U/mL、925 U/mL、965 U/mL。通过对各重组菌上清液酶活力、胞内滞留酶活力、上清液蛋白浓度数据进行分析,进一步选择将HAC1、ERO1、PDI进行两基因或三基因组合,并分别在分泌表达ManA的重组菌胞内共表达,但各共表达重组菌发酵上清液的酶活力都没有进一步的提升。单独共表达HAC1或者分子伴侣ERO1、PDI可以辅助ManA的正确折叠,提高其蛋白表达。展开更多
Three-dimensional finite element models were developed to analyze 304 stainless steel rod and wire hot continuous rolling process with the help of MSC.Marc software. The entire 30-pass deformation process and the actu...Three-dimensional finite element models were developed to analyze 304 stainless steel rod and wire hot continuous rolling process with the help of MSC.Marc software. The entire 30-pass deformation process and the actual parameters of production line were taken into account. Static and dynamic procedures were used to study the continuous rolling process with the aid of the thermo-mechanical coupled FEM of elastic-plasticity. The properties of billets, such as deformation, temperature field and rolling force, were mainly discussed. The simulation results of temperature agree well with the measured values. Comparisons of the analysis results obtained using static implicit method and dynamic implicit method were presented. It is shown that static implicit procedure is more accurate than dynamic implicit procedure and is able to simulate the rolling process with a lower speed, such as a roughing mill. Whereas, dynamic analysis shows a higher efficiency than static analysis and is fit for simulating the rolling process with a higher speed, such as a finishing mill.展开更多
基金supported by the Strategic Priority Research Program of the Chinese Academy of Sciences(No.XDA22010101)the National Natural Science Foundation of China(No.51871225)+1 种基金the National Key Research and Development Program of China(No.2021YFC2801800)the Youth Innovation Promotion Association CAS and Liaoning Revitalization Talents Program,China(No.XLYC1907005).
文摘Three-dimensional finite element models were developed to analyze 304 stainless steel rod and wire hot continuous rolling process with the help of MSC.Marc software. The entire 30-pass deformation process and the actual parameters of production line were taken into account. Static and dynamic procedures were used to study the continuous rolling process with the aid of the thermo-mechanical coupled FEM of elastic-plasticity. The properties of billets, such as deformation, temperature field and rolling force, were mainly discussed. The simulation results of temperature agree well with the measured values. Comparisons of the analysis results obtained using static implicit method and dynamic implicit method were presented. It is shown that static implicit procedure is more accurate than dynamic implicit procedure and is able to simulate the rolling process with a lower speed, such as a roughing mill. Whereas, dynamic analysis shows a higher efficiency than static analysis and is fit for simulating the rolling process with a higher speed, such as a finishing mill.