Abstract: Phospholipase D (PLD, EC 3.1.4.4) is a suite of enzymes catalyzing the hydrolysis of the phosphodiester bond of phospholipids, and the transfer of phosphatidic acid to primary alcohols to generate different phospholipids. The transphosphatidylation reaction makes PLD have wide applications in food, pharmaceutical and cell biology fields. However, large-scale industrial production of PLD has been restricted by many factors. Genetic engineering technology has therefore been adopted to clone and express PLD genes to obtain PLD with high quality, high purity and high transphosphatidylation activity. In this review, current progresses on the cloning and high-efficiency expression of microbial phospholipase D are analyzed, with a hope to guide the large-scale industrial production of PLD and lay a solid foundation for the development and utilization of related products.