P450BM3 enzyme mutant capable of efficiently utilizing non-natural cofactors and expression application of P450BM3 enzyme mutant

The invention provides a P450BM3 enzyme mutant capable of efficiently utilizing non-natural cofactors, which is characterized in that the P450BM3 enzyme mutant capable of efficiently utilizing non-natural cofactors is rationally designed by taking PDB serial numbers 1J9Z and 4DQL as structural templates and taking the distance between N-5 atoms of FAD flavin and C-4 atoms of NMNH as the basis of improving the catalytic efficiency to improve the activity of a bionic cofactor NMNH utilized by a P450BM3 enzyme, and finally, the P450BM3 enzyme mutant capable of efficiently utilizing non-natural cofactors is obtained. The method is applied to a secretory expression system. The P450BM3 enzyme mutant disclosed by the invention can efficiently utilize a bionic cofactor NMNH, so that the cost and obstacles of industrial production can be greatly reduced. 本发明提供了一种能高效利用非天然辅因子的P450BM3酶突变体，以PDB编号1J9Z与4DQL为结构模板，以FAD黄素的N-5原子与NMNH的C-4原子之间的距离为改善催化效率的基础进行理性设计提高P450BM3酶利用仿生辅因子NMNH的活性，最后得到能高效利用非天然辅因子的P450BM3酶突变体，将其应用到了分泌表达系统中。本发