Abstract: The effect of preservation in distilled water on production of glucose oxidase with recombinant Pichia pastoris was investigated in comparision with the method of slant preservation. When the strain Pichia pastoris P51 was preserved with methods of distilled water preservation and slant preservation for 180 days, strain survival rate reached 90% and 45%, respectively, which indicated that the effect of distilled water was better than the latter. Furthermore, the final enyzme activity reached (22.5 ±0.5) U/m L and (23 ±0.4) U/m L under methods of distilled water preservation and slant preservation in flask culture system. It showed that distilled water preservation did not reduce the yield of glucose oxidase. Besides, the distilled water preservation compared to slant preservation had some advantages such as execution at room temperature and simply operation.