定点突变法提高蛋白酶PT121催化合成Z-阿斯巴甜的效率研究任务书
2020-05-22 21:10:30
1. 毕业设计（论文）的内容和要求
论文内容
阿斯巴甜(aspartame，apm)，作为蔗糖的重要替代品，甜度约为蔗糖的200倍，但其热值仅为蔗糖的1/200。已经成为国际市场上的主导强力甜味剂，前期筛选到的来源于pseudomonas aeruginosa 所产蛋白酶pt121具有高效催化阿斯巴甜前体(z-apm)的潜在能力，本课题拟通过定点突变提高催化z-阿斯巴甜合成。
论文要求
2. 参考文献
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[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 Family11 xylanase by directed evolution and sitedirected 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
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起讫日期 |
设计（论文）各阶段工作内容 |
备 注 |
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2015.1. 4-2015.1.24 |
进行文献检索、阅读资料，撰写开题报告 |
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2015.3.1-2015.4.15 |
考察筛选突变位点，考察突变体对合成效率影响 |
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2015.4.15-2015.5.15 |
进一步优化催化体系，调控催化效率至最佳状态 |
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2015.5.15-2015.6.1 |
完善实验结构，整理实验数据，完成毕业论文 |
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2015.6初 |
毕业答辩 |
