Several kinds of organic acids, alcohols, aromatic compounds and sugars as carbon sources were tested in order to produce the cell mass of Rhizopus oryzae which is used in part of food processing or organic acid fermentation. Sodium acetate among them was good enough for carbon source as well as glucose under the concentration of one percent. All nitrogenous substances tested such as ammonium, nitrate or organic nitrogen compounds were well used by this strain of Rhizopus oryzae as nitrogen source. Ammonium sulfate among inorganic nitrogen compounds was most utilized as a nitrogen source in glucose or acetate medium. This strain did not require any growth factors such as yeast extract. The following composition of medium was therefore determined in order to produce the cell mass of Rhizopus oryzae: Na-acetate 1 %, (NH$_4$)$_2$SO$_4$ 0.2%, $K_2$HPO$_4$ 0.05%, MgSO$_4$.7$H_2O$ 0.01%, NaCl 0.01% (PH 5.5). The cell wall of mycelium grown in above medium was lysed optimally at pH 6.5 and 5$0^{circ}C$ by the action of Strepzyme 115-5. On producing protoplast from mycelium by enzymatic action, almost all of the mycelium was damaged after 4hrs of treatment.