Difference between revisions of "Part:BBa K4759002"

Revision as of 05:38, 10 October 2023

camA

a gene coding for protein PdR(CamA), CamA is a putidaredoxin reductase. In the camphor monooxygenase system from Pseudomonas putida, the containing putidaredoxin (Pdx) shuttles electrons between the NADH-dependent putidaredoxin reductase (Pdr) and cytochrome P450(cam).  this sequence has been codon-optimized for expression in Escherichia coli. The P450 enzymes are redox-dependent proteins, through which they source electrons from reducing cofactors to drive their activities. In bacterial systems, the electrostatic interactions between the ferredoxin (Fdx), P450 heme and the reductase domains, as well as the negatively and positively charged amino acids on the Fdx iron-sulfur cluster and P450 proximal site, mediate the conformational changes of the Fdx for electron transfer to P450s. In addition, the substrate binding to P450 induces P450 conformational change to increase its preference for Fdx through electrostatic and steric complementarity . Optimizing protein to protein interactions using different methods to improve the electron transfer efficiency of the P450 system, known as "redox chaperone engineering", is one of the important means of engineering P450s, and fruitful progress has been made. Several studies have confirmed that the combined expression of P450, enzyme with different RPs can achieve the reconstruction and promotion of reactivity. This strategy has been widely used in the bacterial class I P450 system.

Design Notes

this sequence has been codon-optimized for expression in Escherichia coli.

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Usage and Biology

Sequence and Features