Revised equation of enzymatic kinetics and thermodynamic mechanisms for directed evolution of enzymes

2022 ◽  
Author(s):  
Qinyi Zhao
Keyword(s):  
Directed Evolution ◽  
Enzymatic Kinetics ◽  
Kinetics And Thermodynamic

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.

Optimization of Biosensors by Directed Evolution

2003 ◽  
Author(s):  
Carol A. Fierke
Keyword(s):  
Directed Evolution

Directed Evolution of a Selective and Sensitive Serotonin Biosensor Via Machine Learning

2019 ◽  
Author(s):  
Elizabeth Unger ◽  
Jacob Pearson Keller ◽  
Michael Altermatt ◽  
Ruqiang Liang ◽  
Zi Yao ◽  
...  
Keyword(s):  
Machine Learning ◽  
Directed Evolution
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