High-throughput screening methodology for the directed evolution of glycosyltransferases

2006 ◽  
Vol 3 (8) ◽  
pp. 609-614 ◽  
Author(s):  
Amir Aharoni ◽  
Karena Thieme ◽  
Cecilia P C Chiu ◽  
Sabrina Buchini ◽  
Luke L Lairson ◽  
...  
Keyword(s):  
Directed Evolution ◽  
High Throughput ◽  
High Throughput Screening

Directed Evolution of P450 Fatty Acid Decarboxylases via High-Throughput Screening Towards Improved Catalytic Activity

2019 ◽  
Author(s):  
Huifang Xu ◽  
Weinan Liang ◽  
Linlin Ning ◽  
Yuanyuan Jiang ◽  
Wenxia Yang ◽  
...  
Keyword(s):  
Catalytic Activity ◽  
Fatty Acid ◽  
Directed Evolution ◽  
High Throughput ◽  
High Throughput Screening ◽  
Colorimetric Detection ◽  
Screening Method ◽  
Random Mutagenesis ◽  
Direct Production ◽  
Consumption Activity

P450 fatty acid decarboxylases (FADCs) have recently been attracting considerable attention owing to their one-step direct production of industrially important 1-alkenes from biologically abundant feedstock free fatty acids under mild conditions. However, attempts to improve the catalytic activity of FADCs have met with little success. Protein engineering has been limited to selected residues and small mutant libraries due to lack of an effective high-throughput screening (HTS) method. Here, we devise a catalase-deficient <i>Escherichia coli</i> host strain and report an HTS approach based on colorimetric detection of H<sub>2</sub>O<sub>2</sub>-consumption activity of FADCs. Directed evolution enabled by this method has led to effective identification for the first time of improved FADC variants for medium-chain 1-alkene production from both DNA shuffling and random mutagenesis libraries. Advantageously, this screening method can be extended to other enzymes that stoichiometrically utilize H<sub>2</sub>O<sub>2</sub> as co-substrate.

scholarly journals A Photoclick‐Based High‐Throughput Screening for the Directed Evolution of Decarboxylase OleT

2020 ◽  
Author(s):  
Ulrich Markel ◽  
Pia Lanvers ◽  
Daniel F. Sauer ◽  
Malte Wittwer ◽  
Gaurao V. Dhoke ◽  
...  
Keyword(s):  
Directed Evolution ◽  
High Throughput ◽  
High Throughput Screening

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

2013 ◽  
Vol 52 (21) ◽  
pp. 5571-5574 ◽  
Author(s):  
Ryan Lauchli ◽  
Kersten S. Rabe ◽  
Karolina Z. Kalbarczyk ◽  
Amulya Tata ◽  
Thomas Heel ◽  
...  
Keyword(s):  
Directed Evolution ◽  
High Throughput ◽  
High Throughput Screening ◽  
Terpene Synthase

Flow Cytometer-Based High-Throughput Screening System for Accelerated Directed Evolution of P450 Monooxygenases

ACS Catalysis ◽  
2012 ◽  
Vol 2 (12) ◽  
pp. 2724-2728 ◽  
Author(s):  
Anna Joëlle Ruff ◽  
Alexander Dennig ◽  
Georgette Wirtz ◽  
Milan Blanusa ◽  
Ulrich Schwaneberg
Keyword(s):  
Directed Evolution ◽  
High Throughput ◽  
High Throughput Screening ◽  
Flow Cytometer ◽  
Screening System ◽  
P450 Monooxygenases

Design of Modern Biopharmaceuticals by Ultra-high-throughput Screening and Directed Evolution

2008 ◽  
pp. 582-603
Author(s):  
Markus Rarbach ◽  
Wayne M. Coco ◽  
Andre Koltermann ◽  
Ulrich Kettling ◽  
Manfred Eigen
Keyword(s):  
Directed Evolution ◽  
High Throughput ◽  
High Throughput Screening
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