全文总字数：2436字
1. 毕业设计（论文）的内容和要求
内容：以酿酒酵母可分泌的细胞壁蛋白为锚定蛋白，通过在细胞表面展示絮凝类蛋白，提高酿酒酵母结合固定化载体的能力，构建基于生物被膜的表面固定化发酵体系。
要求：（1）筛选出两种锚定蛋白；（2）以GFP作为报告基因，在酿酒酵母表面展示GFP；（3）选定1-2种絮凝类相关的基因，并在细胞表面进行展示；（4）基于细胞表面展示的絮凝类相关基因，构建基于生物被膜的表面固定化发酵体系。
2. 实验内容和要求
1 掌握酿酒酵母表达质粒的构建方法；2 掌握酿酒酵母感受态细胞的制备、电击转化、菌落PCR等实验方法；3 通过不同的表征手段表征酿酒酵母表面展示的效果，并构建基于生物被膜的表面固定化发酵体系；4 学习并掌握整理实验数据、分析实验结果并得到相应的实验结论的能力。
3. 参考文献
1. Tsutomu T , Akihiko K . Cell-surface display of enzymes by the yeast Saccharomyces cerevisiae for synthetic biology.[J]. FEMS Yeast Research(1):1.2. Inokuma K , Hasunuma T , Kondo A . Efficient yeast cell-surface display of exo- and endo-cellulase using the SED1 anchoring region and its original promoter[J]. Biotechnology for Biofuels, 2014, 7(1):1-11.3. Su G D , Zhang X , Lin Y . Surface display of active lipase in Pichia pastoris using Sed1 as an anchor protein[J]. Biotechnology Letters, 2010, 32(8):1131-1136.4. Inokuma K , Bamba T , Ishii J , et al. Enhanced cell-surface display and secretory production of cellulolytic enzymes with Saccharomyces cerevisiae Sed1 signal peptide.[J]. Biotechnology Bioengineering, 2016.5. Matsuoka H , Hashimoto K , Saijo A , et al. Cell wall structure suitable for surface display of proteins in Saccharomyces cerevisiae[J]. Yeast, 2014, 31(2):67-76.6. Zhuo, Liu, Kentaro, et al. Combined cell-surface display- and secretion-based strategies for production of cellulosic ethanol with Saccharomyces cerevisiae[J]. Biotechnology for Biofuels, 2015.7. Tanaka T , Yamada R , Ogino C , et al. Recent developments in yeast cell surface display toward extended applications in biotechnology[J]. Applied Microbiology and Biotechnology, 2012, 95(3):577-591.8. Ueda K M . Cell surface engineering of yeast for applications in white biotechnology[J]. Biotechnology Letters, 2011.9. Liang C , Ding S , Sun W , et al. Biofilm-based fermentation: a novel immobilisation strategy for Saccharomyces cerevisiae cell cycle progression during ethanol production[J]. Applied Microbiology and Biotechnology, 2020, 104(17).10. Tanaka T , Yamada R , Ogino C , et al. Recent developments in yeast cell surface display toward extended applications in biotechnology[J]. Applied Microbiology and Biotechnology, 2012, 95(3):577-591.
4. 毕业设计（论文）计划
2020.12-2021.01 下载并阅读与课题相关的文献，完成开题报告2021.02-2021.03 构建酿酒酵母相关的表达质粒；2021.03-2021.05 转化至酿酒酵母细胞并筛选出重组菌株，并通过不同的表征方法表征表面展示效果；2021.05-2021.06 整理实验数据并撰写毕业论文。
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