Self-sufficient Cytochrome P450s and their potential applications in biotechnology

Cytochrome P450s(CYPs)are ubiquitously found in all kingdoms of life,playing important role in various biosynthetic pathways as well as degradative pathways;accordingly find applications in a vast variety of areas from organic synthesis and drug metabolite production to modification of biomaterials and bioremediation.Significantly,CYPs catalyze chemically challenging CAH and CAC activation reactions using a reactive high-valent iron-oxo intermediate generated upon dioxygen activation at their heme center,while the other oxygen atom is reduced to the level of water by electrons provided through a reductase partner protein.Self-sufficient CYPs,encoding their heme domain and reductase protein in a single polypeptide,facilitate increased catalytic efficiency and render a less complicated system to work with.The self-sufficient CYP enzyme from CYP102A family(CYP102A1,BM3)is among the earliest and most-investigated model enzymes for mechanistic and structural studies as well as for biotechnological applications.An increasing number of self-sufficient CYPs from the same CYP102 family and from other families have also been reported in last decade.In this review,we introduce chemistry and biology of CYPs,followed by an overview of the characteristics of self-sufficient CYPs and representative reactions.Enzyme engineering efforts leading to novel self-sufficient CYP variants that can catalyze synthetically useful natural and non-natural(nature-mimicking)reactions are highlighted.Lastly,the strategy and efforts that aim to circumvent the challenges for improved thermostability,regio-and enantioselectivity,and total turnover number;associated with practical use of self-sufficient CYPs are reviewed.

Identification and characterization of a cytochrome P450 C YP6CX1 putatively associated with insecticide resistance in Bemisia tabaci

The novel full length of cytochrome P450 gene has been isolated in insecticideresistant （named CYP6CXlvl） and -susceptible （named CYP6CXlv2） Bemisia tabaci, which was identified as B biotype, in Shangjie, Fujian, China （Sj）. CYP6CX1 （1 940 bp contained a 1 557 bp open reading frame） included conserved domains common to CYP6 members, such as heme-binding motif PFGEGPRFCIA, putative ＂meander＂-binding sequence ETLR and PERF in helix-K, oxygen-binding motif AGLDPV and conserved sequence PEKFNP near the carboxyl end. There were four different replacements of amino acid residues between R and S B. tabaci （Thr300 Ala, Thr354Pro, Arg486His and Ile503Thr）, among which the substitution Ile503Thr was located in the substrate recognition sites region. The mRNA transcription level of CYP6CXlvl was 2.38-fold as high as that of CYP6CXlv2. The results indicated that the CFP6CX1 from the B biotype B. tabaci in Sj was one of the CYP6 members, and enhanced CYP6CX1 expression and substitute of amino acid residues might be involved in the resistance mechanisms in field B. tabaci.