As important metabolic enzymes, the function of cytochrome P450 monooxygenases (CYPs) has been demonstrated repeatedly through various means, including heterologous expression systems. Unfortunately, most model systems typically lack expression of a conspecific NADPH-dependent cytochrome P450 reductase (CPR), which is the electron transfer partner of CYPs. As a result, the activities of heterologously expressed insect CYPs may not accurately reflect detoxification activities in vivo. Previously, CYP6B7 from Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) (HaCYP6B7) has been expressed in the Pichia pastoris GS115 strain and shown to detoxify bifenthrin, fenvalerate and chlorpyrifos. However, it remains to be determined if co-expression of HaCYP6B7 with HaCPR will enhance the detoxification ability of the expression system. In the present study, HaCYP6B7 and HaCPR genes were co-expressed in P. pastoris using a reconstituted expression vector, pPICZA-HaCYP6B7-HaCPR. Protein expression was confirmed by Western blot, and the detoxification activities of microsomal fractions to p-nitroanisole O-demethylation (PNOD), 7-ethoxycoumarin O-deethylation (ECOD), fenvalerate and chlorpyrifos were measured. Co-expression of HaCYP6B7 with HaCPR resulted in PNOD and ECOD activities of 1.90 nmol/min/mg·protein and 12.39 pmol/min/mg·protein, which were 1.6- and 1.5-fold of that catalyzed by HaCYP6B7 expressed alone, respectively. Furthermore, microsomes of pPICZA-HaCYP6B7-HaCPR-GS115 had higher detoxification activity than that of pPICZA-HaCYP6B7-GS115 to fenvalerate, but not chlorpyrifos. The results indicated that co-expression of HaCYP6B7 with conspecific CPR could enhance the detoxification activities to some substrates comparing with expression of HaCYP6B7 alone.