全文总字数：4235字
1. 毕业设计（论文）的内容和要求
1．文献查阅 掌握文献查阅，使用网络资源如中国期刊网、维普数据库、超星数字图书馆、Elsevier电子期刊、Springer Link全文电子期刊等检索资源，查阅关于塑料降解酶条件优化方面的相关文献。
2．文献阅读及综述 阅读与课题相关的外文与中文文献，了解国内外的研究动态，撰写文献综述。
3．明确实验任务，拟定实验方案 根据所查阅文献的内容，确定实验内容及方案，拟定科学可行的研究计划。
2. 实验内容和要求
本课题通过分子生物学技术，筛选多种信号肽介导PETase实现其在大肠杆菌中的可溶性表达，并对目的蛋白进行纯化，验证其酶活性。
需解决的问题： 1.信号肽OmpA、PelB、MalE与PETase融合构建重组表达载体 2.高效表达条件的优化以及纯化条件的摸索。
3、PETase酶活验证。
3. 参考文献
1. Sinha, V.; Patel, M.; Patel, J., Pet Waste Management by Chemical Recycling: A Review. Journal of Polymers and the Environment 2010, 18, 8-25.2. Fecker, T.; Galaz-Davison, P.; Engelberger, F.; Narui, Y.; Sotomayor, M.; Parra, L. P.; Ramrez-Sarmiento, C. A., Active Site Flexibility as a Hallmark for Efficient PET Degradation by I. sakaiensis PETase. Biophysical journal 2018, 114 (6), 1302-1312.3. Kawai, F.; Kawabata, T.; Oda, M., Current State and Perspectives Related to the Polyethylene Terephthalate Hydrolases Available for Biorecycling. ACS Sustainable Chemistry Todd, A., The thermal degradation of polyethylene terephthalate. Transactions of The Faraday Society 1953, 49.5. Paszun, D.; Spychaj, T., Chemical Recycling of Poly(ethylene terephthalate). Industrial Lorenz, G.; Kandelbauer, A., Recycling of poly (ethylene terephthalate)-A review focusing on chemical methods. eXPRESS Polymer Letters 2016, 10, 559-586.7. Seo, H.; Kim, S.; Son, H.; Sagong, H.-Y.; Joo, S.; Kim, K.-J., Production of extracellular PETase from Ideonella sakaiensis using sec-dependent signal peptides in E. coli. Biochemical and Biophysical Research Communications 2018, 508.8. Kawai, F.; Kawabata, T.; Oda, M. J. S. O. C., Current knowledge on enzymatic PET degradation and its possible application to waste stream management and other fields. 2019, 103 (11).9. Mller, R.-J.; Schrader, H.; Profe, J.; Dresler, K.; Deckwer, W.-D., Enzymatic Degradation of Poly(ethylene terephthalate): Rapid Hydrolyse using a Hydrolase from T. fusca. Macromolecular Rapid Communications 2005, 26 (17), 1400-1405.10. Ronkvist, A.; Xie, W.; Lu, W.; Gross, R., Cutinase-Catalyzed Hydrolysis of Poly(ethylene terephthalate). Macromolecules 2009, 42.11. Silva, C.; Da, S.; Silva, N.; Matam, T.; Arajo, R.; Martins, M.; Chen, S.; Chen, J.; Wu, J.; Casal, M.; Cavaco-Paulo, A., Engineered Thermobifida fusca cutinase with increased activity on polyester substrates. Biotechnology journal 2011, 6, 1230-9.12. Wei, R.; Zimmermann, W., Biocatalysis as a green route for recycling the recalcitrant plastic polyethylene terephthalate. Microbial Biotechnology 2017, 10 (6), 1302-1307.13. Castro, A.; Carniel, A.; Junior, J.; Gomes, A.; Valoni, ., Screening of commercial enzymes for poly(ethylene terephthalate) (PET) hydrolysis and synergy studies on different substrate sources. Journal of Industrial Microbiology Tong, X.; Woodard, R.; Du, G.; Chen, J., Identification and Characterization of Bacterial Cutinase. The Journal of biological chemistry 2008, 283, 25854-62.15. Then, J.; Wei, R.; Oeser, T.; Barth, M.; Belisrio-Ferrari, M.; Schmidt, J.; Zimmermann, W., Ca2 and Mg2 binding site engineering increases the degradation of polyethylene terephthalate films by polyester hydrolases from Thermobifida fusca. Biotechnology Journal 2015, 10, 592-598.16. Herrero Acero, E.; Ribitsch, D.; Dellacher, A.; Zitzenbacher, S.; Marold, A.; Steinkellner, G.; Gruber, K.; Schwab, H.; Guebitz, G., Surface Engineering of a Cutinase From Thermobifida Cellulosilytica for Improved Polyester Hydrolysis. Biotechnology and bioengineering 2013, 110.17. Yoshida, S.; Hiraga, K.; Takehana, T.; Taniguchi, I.; Yamaji, H.; Maeda, Y.; Toyohara, K.; Miyamoto, K.; Kimura, Y.; Oda, K., A bacterium that degrades and assimilates poly(ethylene terephthalate). Science 2016, 351, 1196-1199.
4. 毕业设计（论文）计划
2021.1-2021.2 文献查阅，完成开题报告，熟悉实验仪器。
2021.3-2021.4 重组表达载体的构建 2021.4-2021.5 高效表达条件的优化以及纯化条 件的摸索 2021.5-2021.6 PETase纯化及酶活鉴定 2021.6上旬 完成论文及答辩
以上是毕业论文任务书，课题毕业论文、开题报告、外文翻译、程序设计、图纸设计等资料可联系客服协助查找。
