Carpenter J F, Martin B, Loomis S H, Crowe J H
Department of Zoology, University of California, Davis 95616.
Cryobiology. 1988 Aug;25(4):372-6. doi: 10.1016/0011-2240(88)90045-4.
We have demonstrated that sugars and suger/zinc mixtures can be used to preserve the activity of dried phosphofructokinase (PFK) during long-term storage over CaSO4. After 9 weeks in the presence of either 200 mM sucrose or 200 mM trehalose little loss of PFK activity was noted, with almost 60% of the original prefreeze-dry activity recovered when samples were rehydrated. Even reducing sugars protected the dried enzyme throughout the entire storage period. Of the sugars tested, 200 mM lactose provided the most stability to PFK; at the end of the dry storage, over 80% of the initial activity was recovered. With either 200 mM maltose or 400 mM glucose, about 40% of the initial activity was recovered at the end of the experiment. With all the sugars tested, the addition of 0.6 mM Zn2+ to sugar/PFK mixtures enhanced the stability of the enzyme, and no long-term adverse effects of the metal ion on enzyme activity were noted.
我们已经证明,糖类和糖/锌混合物可用于在硫酸钙上长期储存期间保存干燥的磷酸果糖激酶(PFK)的活性。在存在200 mM蔗糖或200 mM海藻糖的情况下9周后,未观察到PFK活性有明显损失,当样品复水时,几乎恢复了60%的冻干前原始活性。即使是还原糖在整个储存期间也能保护干燥的酶。在所测试的糖类中,200 mM乳糖为PFK提供了最大的稳定性;在干燥储存结束时,恢复了超过80%的初始活性。使用200 mM麦芽糖或400 mM葡萄糖时,在实验结束时恢复了约40%的初始活性。对于所有测试的糖类,向糖/PFK混合物中添加0.6 mM Zn2+可增强酶的稳定性,并且未观察到金属离子对酶活性有长期不利影响。