High-throughput screening for terpene-synthase-cyclization activity and directed evolution of a terpene synthase.

High-throughput screening for terpene-synthase-cyclization activity and directed evolution of a terpene synthase.
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DOI:
10.1002/anie.201301362
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发表时间:
2013-05-17
影响因子:
16.6
通讯作者:
Arnold, Frances H.
Arnold, Frances H.
中科院分区:
化学1区
文献类型:
--
作者:
Lauchli, Ryan;Rabe, Kersten S.;Kalbarczyk, Karolina Z.;Tata, Amulya;Heel, Thomas;Kitto, Rebekah Z.;Arnold, Frances H.

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一种合成底物使得能够对萜烯合成酶环化活性进行新的比色筛选,从而促进这些酶的工程。通过定向进化,倍半萜合成酶BcBOT2的热稳定性在不损失其他性质的情况下得到了提高。该技术还可以快速优化另一种萜烯合酶SSCG_02150裂解产物的表达和稳定条件。
A synthetic substrate enables a new colorimetric screen for terpene synthase cyclization activity, facilitating the engineering of these enzymes. Using directed evolution, the thermostability of the sesquiterpene synthase BcBOT2 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, SSCG_02150.
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