The invention discloses bacillus subtilis with inactivated extracellular protease as well as a construction method and application the bacillus subtilis. B.subtilis 168 is taken as an original strain, a transcription factor comK-comS gene controlled by a xylose inducible promoter PxylA is integrated on a genome by using a CRISPR/Cpf1 technology, so that the transformation efficiency of the bacillus subtilis is greatly improved; a trpC gene and a gudB gene are subjected to reverse mutation, so that tryptophan can be synthesized, and glutamic acid family amino acids can be effectively utilized; and besides, six extracellular protease genes are knocked out, so that the metabolic pressure of cells is reduced, and recombinant proteins can be effectively accumulated outside the cells. The content of extracellular sfGFP produced when a final engineering strain G601 is used for fermentation is 1.88 times of that of the B.subtilis 168, and the total content of intracellular and extracellular sfGFP is 1.94 times of that of the B.subtilis 168. An engineering bacterium constructed by the invention is simple and convenient to transform, has no nutritional deficiency, has better amino acid utilization capacity and low extracellular protease activity, can be widely applied to secretory expression of a recombinant protein, and has a wide application prospect.