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毕业论文网 > 任务书 > 化学化工与生命科学类 > 制药工程 > 正文

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 09:05  

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

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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 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

起讫日期

设计(论文)各阶段工作内容

备 注

2015.1. 4-2015.1.24

进行文献检索、阅读资料,撰写开题报告

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

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

2015.5.15-2015.6.1

完善实验结构,整理实验数据,完成毕业论文

2015.6初

毕业答辩

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