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The experiment was to screen strains to produce protease by fermenting silk reeling wastewater. The strain Z416 was screened by sampling from natural environment. The strain was identified as Paenarthrobacter nitroguajacolicus by morphological observation and molecular biology. The experiment further optimized the combination of enzyme production conditions of strain through single factor and orthogonal design experiments. The results showed that the optimal fermentation scheme was lactose addition amount of 4% in fermentation broth, initial pH value of fermentation medium of 7.5, inoculum amount of 16%, the speed of the shaking table of 180 r/min, fermentation temperature of 33 ℃ and fermentation time of 84 h. Under these conditions, the enzyme activity of strain Z416 reached 17.89 U/mL, which was 5.57 times that of 3.21 U/mL before optimization. The study indicates the protease has the potential to be used in poultry and pig feed to improve the growth performance of livestock and poultry. [ABSTRACT FROM AUTHOR]