Hill D J, Jenkins R O, Cartledge T G, Lloyd D
Biochem J. 1986 Aug 15;238(1):255-61. doi: 10.1042/bj2380255.
Adaptation of methanol-grown Candida boidinii to ethanol utilization was accompanied by an increase in proteolytic activities, which behaved like known vacuolar enzymes. Degradation of alcohol oxidase protein was partially prevented by the serine proteinase inhibitor phenylmethanesulphonyl fluoride, but not by the carboxyl proteinase inhibitor pepstatin. Fractionation of cell-free extracts, by high-speed zonal centrifugation, of methanol-grown C. boidinii showed non-sedimentable and sedimentable proteolytic activities. Naturally occurring inhibitors of vacuolar proteinases were non-sedimentable. Fractionation of extracts prepared from methanol-grown cells which had been adapted to ethanol utilization for 5 h revealed significant changes in the sedimentability and distribution of proteolytic and acid phosphatase activities. These results suggest the possible involvement of a vacuolar process during alcohol oxidase degradation.
甲醇培养的博伊丁假丝酵母适应乙醇利用的过程伴随着蛋白水解活性的增加，这些活性表现得类似于已知的液泡酶。丝氨酸蛋白酶抑制剂苯甲基磺酰氟可部分阻止乙醇氧化酶蛋白的降解，但羧基蛋白酶抑制剂胃蛋白酶抑制剂则不能。通过高速区带离心对甲醇培养的博伊丁假丝酵母的无细胞提取物进行分级分离，显示出不可沉淀和可沉淀的蛋白水解活性。液泡蛋白酶的天然抑制剂是不可沉淀的。对适应乙醇利用5小时的甲醇培养细胞制备的提取物进行分级分离，发现蛋白水解和酸性磷酸酶活性的沉淀性和分布发生了显著变化。这些结果表明在乙醇氧化酶降解过程中可能涉及液泡过程。
