High-Throughput Screening for Terpene-Synthase-Cyclization Activity and Directed Evolution of a Terpene Synthase

被引：72
作者：
Lauchli, Ryan [1 ]
Rabe, Kersten S. [1 ]
Kalbarczyk, Karolina Z. [1 ]
Tata, Amulya [1 ]
Heel, Thomas [1 ]
Kitto, Rebekah Z. [1 ]
Arnold, Frances H. [1 ]
机构：
[1] CALTECH, Div Chem & Chem Engn, Pasadena, CA 91125 USA
来源：
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION | 2013年 / 52卷 / 21期
基金：
奥地利科学基金会;
关键词：
directed evolution; high-throughput screening; protein engineering; terpenes; thermostability; FARNESYL DIPHOSPHATE; ALCOHOL OXIDASE; STREPTOMYCES-COELICOLOR; SESQUITERPENE SYNTHASES; ARISTOLOCHENE SYNTHASE; METHANOL; ASSAY; OVERPRODUCTION; BIOSYNTHESIS; FORMALDEHYDE;
D O I：
10.1002/anie.201301362
中图分类号：
O6 [化学];
学科分类号：
0703 ;
摘要：
An easy assay: A synthetic substrate enables a colorimetric screen for terpene synthase cyclization activity, thereby facilitating the engineering of these enzymes. By using directed evolution, the thermostability of a sesquiterpene synthase was increased without the loss of other properties. The technique also enabled rapid optimization of conditions for expression and stabilization in lysate of another terpene synthase. PPO=diphosphate. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
引用
收藏
页码：5571 / 5574
页数：4
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