A5041733718- Microbial Metabolic Engineering and Bioproduction
- Enzyme Catalysis and Immobilization
- Biochemical and Molecular Research
- CRISPR and Genetic Engineering
- Photosynthetic Processes and Mechanisms
- Mitochondrial Function and Pathology
- Enzyme Structure and Function
- RNA and protein synthesis mechanisms
- Bacterial Genetics and Biotechnology
- Microbial bioremediation and biosurfactants
- ATP Synthase and ATPases Research
- Protein Structure and Dynamics
- Microbial Natural Products and Biosynthesis
- Electrochemical sensors and biosensors
University of California, Irvine
2020-2024
Abstract Background Noncanonical redox cofactors are emerging as important tools in cell-free biosynthesis to increase the economic viability, enable exquisite control, and expand range of chemistries accessible. However, these noncanonical need be biologically synthesized achieve full integration with renewable biomanufacturing processes. Results In this work, we engineered Escherichia coli cells biosynthesize cofactor nicotinamide mononucleotide (NMN + ), which has been efficiently used...
We report an aerobic, growth-based selection platform founded on NADP(H) redox balance restoration in Escherichia coli, and we demonstrate its application the high-throughput evolution of oxygenase. A single round followed by a facile growth assay enabled Pseudomonas aeruginosa 4-hydroxybenzoate hydroxylase (PobA) to efficiently hydroxylate both 4-hydroxybenzoic acid (4-HBA) 3,4-dihydroxybenzoic (3,4-DHBA), two consecutive steps gallic biosynthesis. Structural modeling suggests precise...
Noncanonical cofactors such as nicotinamide mononucleotide (NMN
Cyclohexanone monooxygenases (CHMO) consume molecular oxygen and NADPH to catalyze the valuable oxidation of cyclic ketones. However, CHMO usage is restricted by poor stability stringent specificity for NADPH. Efforts engineer have been limited sensitivity enzyme perturbations in conformational dynamics long-range interactions that cannot be predicted. We demonstrate an aerobic, high-throughput growth selection platform Escherichia coli oxygenase evolution based on NADH redox balance....
Natural metabolism relies on chemical compartmentalization of two redox cofactors, NAD
The future of biomanufacturing is dependent on rewiring biological systems to establish an alternative approach our current chemical industries. However, a key limitation in that desired processes must rely the same two redox cofactors as natural metabolism, nicotinamide adenine dinucleotide (phosphate) NAD(P)+, shuttle electrons energy. Thus, competition resources with reactions within host cells nearly unavoidable. One strategy overcome cofactor resource implementation third, noncanonical...
ABSTRACT We report an aerobic, growth-based selection platform founded on NADP(H) redox balance restoration in Escherichia coli , and demonstrate its application high-throughput evolution of oxygenase. A single round enabled Pseudomonas aeruginoasa 4-hydroxybenzoate hydroxylase (PobA) to accept 3,4-dihydroxybenzoic acid efficiently, essential step toward gallic biosynthesis. The best variant DA015 exhibited more than 5-fold higher catalytic efficiency compared previously engineered enzymes....
ABSTRACT Cyclohexanone monooxygenases (CHMO) consume molecular oxygen and NADPH to catalyze the valuable oxidation of cyclic ketones. However, CHMO usage is restricted by poor thermostability stringent specificity for NADPH. Efforts engineer have been limited sensitivity enzyme perturbations in conformational dynamics long-range interactions that cannot be predicted. We demonstrate a pair aerobic, high-throughput growth selection platforms Escherichia coli oxygenase evolution, based on or...
Abstract Background Noncanonical redox cofactors are emerging as important tools in cell-free biosynthesis to increase the economic viability, enable exquisite control, and expand range of chemistries accessible. However, these noncanonical need be biologically synthesized achieve full integration with renewable biomanufacturing processes. Results In this work, we engineered Escherichia coli cells biosynthesize cofactor nicotinamide mononucleotide (NMN + ), which has been efficiently used...