Enhancing the Thermostability of Feruloyl Esterase EstF27 by Directed Evolution and the Underlying Structural Basis
Cao, Li-chuang1; Chen, Ran1; Xie, Wei1; Liu, Yu-huan1; xiewei6@mail.sysu.edu.cn; lsslyh@mail.sysu.edu.cn
2015
发表期刊JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
卷号63期号:37页码:8225-8233
摘要To improve the thermostability of EstF27, two rounds of random mutagenesis were performed. A thermostable mutant, M6, with six amino acid substitutions was obtained. The half-life of M6 at 55 degrees C is 1680 h, while that of EstF27 is 0.5 h. The K-cat/K-m value of M6 is 1.9-fold higher than that of EstF27. The concentrations of ferulic acid released from destarched wheat bran by EstF27 and M6 at their respective optimal temperatures were 223.2 +/- 6.8 and 464.8 +/- 11.9 mu M, respectively. To further understand the structural basis of the enhanced thermostability, the crystal structure of M6 is determined at 2.0 angstrom. Structural analysis shows that a new disulfide bond and hydrophobic interactions formed by the mutations may play an important role in stabilizing the protein. This study not only provides us with a robust catalyst, but also enriches our knowledge about the structure-function relationship of feruloyl esterase.
部门归属LMB
关键词Feruloyl Esterase Directed Evolution Thermostability Wheat Bran Crystal Structure X-ray Crystallography
学科领域Agriculture ; Chemistry ; Food Science & Technology
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文献类型期刊论文
条目标识符http://ir.scsio.ac.cn/handle/344004/14768
专题中科院海洋生物资源可持续利用重点实验室
通讯作者xiewei6@mail.sysu.edu.cn; lsslyh@mail.sysu.edu.cn
作者单位1.[Cao, Li-chuang
2.Chen, Ran
3.Xie, Wei
4.Liu, Yu-huan] Sun Yat Sen Univ, Sch Life Sci, Guangzhou 510275, Guangdong, Peoples R China
5.[Chen, Ran
6.Xie, Wei] Sun Yat Sen Univ, Sch Life Sci, State Key Lab Biocontrol, Guangzhou 510275, Guangdong, Peoples R China
7.[Cao, Li-chuang
8.Liu, Yu-huan] Sun Yat Sen Univ, South China Sea Bioresource Exploitat & Utilizat, Guangzhou 510275, Guangdong, Peoples R China
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Cao, Li-chuang,Chen, Ran,Xie, Wei,et al. Enhancing the Thermostability of Feruloyl Esterase EstF27 by Directed Evolution and the Underlying Structural Basis[J]. JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY,2015,63(37):8225-8233.
APA Cao, Li-chuang,Chen, Ran,Xie, Wei,Liu, Yu-huan,xiewei6@mail.sysu.edu.cn,&lsslyh@mail.sysu.edu.cn.(2015).Enhancing the Thermostability of Feruloyl Esterase EstF27 by Directed Evolution and the Underlying Structural Basis.JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY,63(37),8225-8233.
MLA Cao, Li-chuang,et al."Enhancing the Thermostability of Feruloyl Esterase EstF27 by Directed Evolution and the Underlying Structural Basis".JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 63.37(2015):8225-8233.
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