Abstract: The Streptomyces sp.CA-1 as purpose strain, by single- factor experiment and orthogonal experiment, the fermentation conditions for phospholipase D production were optimized. Results showed that the optimal fermentation conditions for producing enzyme: glucose 20g /L, beef extract 8.75g /L + fishmeal peptone 8.75g /L, twain 20 32.5g /L, calcium chloride 2.5g /L, inoculums 1.5%, medium initial pH 7.0, culture temperature 30℃, incubation time 3d.Under the best fermentation conditions, the enzyme activity could reach 1605.9U /L, which was7.72 times than that of unoptimized conditions.