Difference between revisions of "Part:BBa K4759202"

Revision as of 07:03, 10 October 2023

T7-RBS3-OleP

P450 enzyme CYP107D1 derived from Streptomyces, abbreviated as Olep, is selected to synthesize the above gene by adding histidine tags at the C-terminus through E. coli codon optimization and subcloning the above genes onto the plasmid pET-28a to obtain the recombinant plasmid pET-28a-oleP.

Usage and Biology

OleP, the cytochrome P450 epoxidase from Streptomyces antibioticus involved in Oleandomycin biosynthesis: functional analysis and crystallographic structure in complex with clotrimazole. PDB DOI: https://doi.org/10.2210/pdb4XE3/pdb Classification: OXIDOREDUCTASE Organism(s): Streptomyces antibioticus Expression System: Escherichia coli BL21(DE3)

we selected high-copy plasmids pRSFDuet, medium-high copy plasmids pETDuet and pET28a, and low-copy plasmid pACYCDuet as carriers to express Olep in E. coli, and the results showed that the recombinant strains constructed with high-copy plasmid pRSFDuet as the carrier had the highest expression of p450 enzyme, cell growth rate, and substrate conversion rate.

Fig1: Selection of suitable plasmid. WT represents E. coli O1 strain. The blue-filled triangle represents the biomass (OD600). The red hollow triangle represents the conversion rate (%). Values and triangles represent the means and standard deviations of biological triplicates. Sequence and Features