We present the fabrication and properties of lactate biosensors. The novel feature is the use of polymerized flavin adenine dinucleotide (FAD) as mediator for electron transfer. The biosensors were prepared using lactate dehydrogenase (LDH), lactate oxidase (LOX), or baker's yeast (BY) immobilized at the surface of the electrode. The sensors using purified enzymes showed good sensitivity, linearity, and stability. The sensitivity of the BY electrodes was slightly lower. The advantages of this type of sensors are discussed in connection with potential applications.