| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | C. tropicalis ATCC 20336 was naturally isolated from an oil field and used as the source of the CYP52A17LL gene (Craft et al., 2003). The CYP52A17LL open reading frame (ORF) sequence (GenBank accession number AY230504) was synthesized with an optimized codon frequency and inserted in the plasmid vector pUC57-Kan (GenScript, USA). The formation of pYeDP60 has been described elsewhere (Pompon et al., 1996). The pPICZA vector was purchased from Invitrogen (USA). Escherichia coli DH5α, S. cerevisiae BY(2R) (equivalent to W(R) (Truan et al., 1993) with the BY4741 genetic background and a supplementary copy of the CPR gene under its own promoter), and P. pastoris X-33 (Invitrogen, USA) were used as the host cells for plasmid propagation and expression of the target gene, respectively. The primers used for gene construction and cloning are shown in Table 1. | Get A Quote |
1,12-Dodecanedioic acid (DDA), a primary compound and an intermediate precursor for various chemical products, is normally produced by chemical synthesis, which presents potential disadvantages. Instead, the biosynthesis of 1,12-DDA by recombinant (r) microorganisms may offer a viable production route. Cytochrome P450 (CYP) can terminally oxidize fatty acids to ω-hydroxy-fatty acids and further to dicarboxylic acids (ω-oxidation). The wild type (r) CYP52A17LL and its engineered L261S/L490S form, in which the two leucine residues were changed into serine (rCYP52A17SS), were expressed in Pichia pastoris and a strain of Saccharomycescerevisiae coexpressing the yeast NADPH cytochrome P450 reductas... More
