1. 毕业设计（论文）的内容和要求

论文内容 蛋白酶是一类重要酶类，用于洗涤和工业催化等领域。

嗜热蛋白酶家族蛋白酶是重要的工业酶制剂，并可以用来催化合成多种生物活性小肽。

然而，作为重要的工业化嗜热蛋白酶酶家族蛋白酶的蛋白表达量至今仍然较低，生产成本昂贵；本课题拟通过表达体系选择，构建高效的表达体系；表达策略优化调控提高嗜热蛋白酶的异源表达水平。

2. 参考文献

[1] Liu LW , Wang B , Chen HG, Wang SY, Wang MD , Zhang SM,Song AD, Shen JW, Wu K, Jia XC. RationalpH - engineering of the thermostable xylanase bas ed on computational model. Process Biochemistry 44 (2009) 912#8211;915. [2] Zhan DL, Han WW, Feng Y. Experimental and computational studies indicate the mutation of Glu12 to increase the thermostability of oligomeric protease from Pyrococcushorikoshii. J Mol Model, 2011; 17(6): 1241-9. [3] Tian J, Wang P, Gao S, Chu X, Wu N, Fan Y. Enhanced thermostability of methyl parathion hydrolase from Ochrobactrum sp. M231 by rational engineering of a glycine to proline mutation. FEBS Journal 2010; 277(23), 4901#8211;8. [4] 唐啸宇,孙洪林,何冰芳. 耐有机溶剂微生物及酶类在生物催化中的应用[J]. 化学进展,2009, (12):2726-2733. [5] Schmid A, Dordick J S, Hauer B, et al. Industrial biocatalysis today and tomorrow[J]. Nature 2001, 409: 258-268. [6] 郭勇. 酶工程[M].第三版.科学出版社：2009 [7] Kumar D , Bhalla T C. Microbial proteases in peptide synthesis: approaches and applications [J]. Appl. Microbiol. Biotechnol. 2005, 68: 726-736. [8] 宋 欣. 微生物酶转化技术［M]. 北京：化学工业出版社.2004,138-183. [9]Liu YH , Lu FP , Li Y , Wang JL , Gao C. Acid stabilization of Bacillus licheniformis alpha amylase through introduction of mutations. Appl Microbiol Biotechnol (2008) 80:795#8211;803. [10] Zheng HC , Liu YH , Sun MZ, Han Y ,Wang JL ,Sun JS ,Lu FP. Improvement of alkali stability and thermostability of Paenibacillus campinasensis Family 11 xylanase by directed evolution and site directed mutagenesis. J Ind Microbiol Biotechnol (2014) 41:153#8211;162 [11] Liu YH, Hu B, Xu YJ, Bo JX ,Fan S, WangJL, Lu FP. Impro vement of the acid stability of Bacillus licheniformis alpha amylase by erro r-prone PCR. Journal of Applied Microbiology ISSN 1364-5072 [12] Hwan Kim S, Subarna Pokhrel, Je Yoo Y. Mutation of non-conserve d amino acids surrounding catalytic site to shif t pH optimum of Bacillus circulans xylanase. Journal of Molecular Catalysis B: Enzymatic 55 (20 08) 130#8211;136. [13]Tim B, J. Joye I, A. Delcour J and M. Courtin C [14] Deng ZM , Yang HQ , Li JH , Shin HD, Du GC, Liu L , ChenJ. Structure-based engineering of alkaline α-amylase from alkaliphilic Alkalimonas amylolytica for improved thermostability. Appl Microbiol Biotechnol (2014) 98:3997#8211;4007 [15] Supl atov D, Panin N, Kirilin E, a Shcherb akovaT, l Kudryavt sev P ,Sˇvedas* V. Compu tational Design of a pH Stable Enzyme:Unde rstanding Molecular Mechanism of Penicillin Acylas e#8217;s Adaptation to Alkaline Conditions.

3. 毕业设计（论文）进程安排

起讫日期 设计（论文）各阶段工作内容 备 注 2017.1. 5-2017.1.24 进行文献检索、阅读资料，撰写开题报告 2017.3.1-2017.4.15 构建重组质粒，并转化至大肠杆菌中 2017.4.15-2017.5.15 发酵重组菌，考察不同条件对重组菌产酶影响 2017.5.15-2017.6.1 完善实验结构，整理实验数据，完成毕业论文 2017.6初 毕业答辩