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

页码：5571 / 5574

页数：4

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