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以1株耻垢分枝杆菌（Mycobacterium smegmatis）为出发菌株,对其产乳酸氧化酶的培养基和发酵条件进行研究.结果表明,培养基中添加3%甘油,0.4%蛋白胨,1%DL-乳酸,0.005%VB1,0.05%KH2PO4·H2O,0.05%MgSO4·7H2O,0.01%硫酸亚铁胺有助于产酶;优化培养条件为：250 mL三角瓶中装液量50 mL,起始pH7.2,培养温度37℃,静置培养4 d,乳酸氧化酶平均酶活为65.4 U/mL.