AI Article Synopsis
- The study introduces a new aerobic selection platform that utilizes NADP(H) redox balance for high-throughput evolution of enzymes.
- Researchers demonstrated this approach by successfully evolving the enzyme 4-hydroxybenzoate hydroxylase (PobA) to efficiently process two important compounds in gallic acid synthesis.
- The findings suggest structural changes in the enzyme's active site are crucial for its improved function, and the platform holds potential for broader use in engineering similar enzymes.
Article Abstract
We report an aerobic, growth-based selection platform founded on NADP(H) redox balance restoration in , and we demonstrate its application in the high-throughput evolution of an oxygenase. A single round of selection followed by a facile growth assay enabled 4-hydroxybenzoate hydroxylase (PobA) to efficiently hydroxylate both 4-hydroxybenzoic acid (4-HBA) and 3,4-dihydroxybenzoic acid (3,4-DHBA), two consecutive steps in gallic acid biosynthesis. Structural modeling suggests precise reorganization of active site hydrogen bond network, which is difficult to obtain without deep navigation of combinatorial sequence space. We envision universal application of this selection platform in engineering NADPH-dependent oxidoreductases.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8294663 | PMC |
| http://dx.doi.org/10.1021/acscatal.0c01892 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!