Part:BBa_K4759203

T7-RBS4-OleP

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.

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

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 102
  Illegal AgeI site found at 200
  
• 1000
  COMPATIBLE WITH RFC[1000]