High-throughput screening of thermostable esterases for industrial bioconversions

被引:41
作者
Lagarde, D
Nguyen, HK
Ravot, G
Wahler, D
Reymond, JL
Hills, G
Veit, T
Lefevre, F
机构
[1] Proteus SA, F-30000 Nimes, France
[2] Univ Bern, Dept Chem & Biochem, CH-3012 Bern, Switzerland
[3] Degussa AG, D-45127 Essen, Germany
关键词
D O I
10.1021/op020019h
中图分类号
O69 [应用化学];
学科分类号
081704 ;
摘要
Thermostable esterases are used in biocatalysis for the synthesis of a variety of products, for example, flavour esters, emulsifiers, or pharmaceutical intermediates. Industrial enzymatic conversions to obtain chemo-, regio-, and stereoselective transformations may require thermostable enzymes active in organic solvents at high temperatures. To find novel enzymes for industrial bioconversions, we have used a new generation of thermostable reporter substrates (CLIPS-O for Catalyst Identification ProcesS per Oxidization) to screen for esterase activity in thermophilic microorganisms belonging to various thermophilic genus. One of the strains (belonging to Thermus genus) exhibiting activity on short- and long-chain ester CLIPS-O substrates has been selected. The genomic library of this strain was screened using a high-throughput functional technology. Positive clones expressing esterases active at high temperature were immediately characterized during screening for chain-length specificity.
引用
收藏
页码:441 / 445
页数:5
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