Ingram E, Holland K T, Gowland G, Cunliffe W J
J Appl Bacteriol. 1983 Apr;54(2):263-71. doi: 10.1111/j.1365-2672.1983.tb02616.x.
Of several commercial media tested, trypticase soya both containing 0.4% (w/v) D-sorbitol was superior as a growth medium for the production of extracellular proteinase by Propionibacterium acnes (strain P-37). Extracellular proteinase, production of which was shown to be growth-associated by both batch and continuous culture studies, was partially purified by 70% (NH4)2SO4 saturation. Sephadex G-75 chromatography and ion exchange on DEAE-Sephadex A-50. It was shown to be a heterogeneous mixture of at least three molecular species of enzyme. Proteinase I was inhibited by EDTA (10(-3) mol/l) and PMSF (5 millimol/l) and stimulated by CaCl2 (190% at 10(-3) mol/l). It had a molecular weight of 20 to 30000 and a broad pH optimum from 6.5 to 7.5. Proteinase II was an alkaline proteinase with a molecular weight of 30 to 40000 which was not significantly inhibited by EDTA (10(-2) mol/l) nor stimulated by CaCl2. Proteinase III represented a minor proportion of the recovered proteolytic activity, had a molecular weight of 20 to 30000 and was most active in the alkaline pH range. This enzyme was inhibited by EDTA (10(-4) mol/l) and PMSF (5 millimol/l), and stimulated by CaCl2 (250% at 10(-2) mol/l).
在所测试的几种商业培养基中，含有0.4%（w/v）D - 山梨醇的胰蛋白胨大豆培养基作为痤疮丙酸杆菌（菌株P - 37）产生细胞外蛋白酶的生长培养基更为优越。通过分批培养和连续培养研究表明，细胞外蛋白酶的产生与生长相关，通过70%硫酸铵饱和度沉淀、Sephadex G - 75柱层析和DEAE - Sephadex A - 50离子交换进行部分纯化。结果表明它是至少三种分子形式的酶的异质混合物。蛋白酶I受EDTA（10⁻³mol/L）和PMSF（5mmol/L）抑制，并受CaCl₂（10⁻³mol/L时为190%）刺激。其分子量为20至30000，最适pH范围较宽，为6.5至7.5。蛋白酶II是一种碱性蛋白酶，分子量为30至40000，不受EDTA（10⁻²mol/L）显著抑制，也不受CaCl₂刺激。蛋白酶III在回收的蛋白水解活性中占比较小，分子量为20至30000，在碱性pH范围内活性最高。该酶受EDTA（10⁻⁴mol/L）和PMSF（5mmol/L）抑制，并受CaCl₂（10⁻²mol/L时为250%）刺激。
