Enzyme and Microbial Technology 2012-07-15

Two plate-based colorimetric assays for screening α-amino acid ester hydrolase with high synthesis/hydrolysis ratio.

Lu Wang, Li-Juan Ye, Yue Pan, Yi Cao

文献索引:Enzyme Microb. Technol. 51(2) , 107-12, (2012)

全文:HTML全文

摘要

α-Amino acid ester hydrolases (AEHs) are enzymes of interest to the semi-synthesis of β-lactam antibiotics with α-amino, such as cephalexin and cefaclor. An undesired side reaction, the hydrolysis of α-amino acid ester, had hindered applications in antibiotics synthesis. Although the enzymes' S/H ratio can be increased by protein engineering, such approaches require a suitable screening assay. Such a screening assay has not yet been described for AEHs. In this paper, we report a 96-well plate format screening procedure for AEHs based on two spectrophotometric assays. To reduce the hydrolysis reaction while maintaining synthesis activity, and to evaluate the effectiveness of the screening strategy, we introduced random mutations in part of the aeh gene from Xanthomonas rubrillineans by error-prone PCR. By a parallel plate-based screening strategy, three mutants with improved S/H ratio, R87L, T132N and N219I, were obtained.Copyright © 2012 Elsevier Inc. All rights reserved.


相关化合物

  • 头孢克罗
  • (|S|)-(+)-α-苯甘氨...
  • D-(-)-α-苯基甘氨酸
  • DL-苯甘氨酸

相关文献:

Cheminformatics analysis of assertions mined from literature that describe drug-induced liver injury in different species.

2010-01-01

[Chem. Res. Toxicol. 23 , 171-83, (2010)]

Translating clinical findings into knowledge in drug safety evaluation--drug induced liver injury prediction system (DILIps).

2011-12-01

[J. Sci. Ind. Res. 65(10) , 808, (2006)]

Chemical genetics reveals a complex functional ground state of neural stem cells.

2007-05-01

[Nat. Chem. Biol. 3(5) , 268-273, (2007)]

Hologram QSAR model for the prediction of human oral bioavailability.

2007-12-15

[Bioorg. Med. Chem. 15 , 7738-45, (2007)]

Quantitative structure-activity relationship and complex network approach to monoamine oxidase A and B inhibitors.

2008-11-13

[J. Med. Chem. 51 , 6740-51, (2008)]

更多文献...