Histidine ammonia-lyase (EC 4.3.1.3) activity in the cell-free extracts of Hartmannella culbertsoni has been partially purified and the optimum activity is found at pH 9.0--9.2. 2. The enzyme required sulphydryl groups for its activity. L-2-Thiohistidine and EDTA competitively inhibit the enzyme. 3. Its molecular weight, as determined by gel filtration technique, is 131,800 daltons and the energy of activation for this enzyme is 15,205 cals/mole. 4. Certain amoebicidal drug and divalent cations have marked inhibitory effect on the enzyme. Co2+ has a profound stimulatory effect.