Patching J W, Rose A H
J Bacteriol. 1971 Oct;108(1):451-8. doi: 10.1128/jb.108.1.451-458.1971.
Saccharomyces cerevisiae NCYC 366 is susceptible to cold osmotic shock. Exponentially growing cells from batch cultures grown in defined medium at 30 C, after being suspended in 0.8 m mannitol containing 10 mm ethylenedia-minetetraacetic acid and then resuspended in ice-cold 0.5 mm MgCl(2), accumulated the nonmetabolizable solutes d-glucosamine-hydrochloride and 2-aminoisobutyrate at slower rates than unshocked cells; shocked cells retained their viability. Storage of unshocked batch-grown cells in buffer at 10 C led to an increase in ability to accumulate glucosamine, and further experiments were confined to cells grown in a chemostat under conditions of glucose limitation, thereby obviating the need for storing cells before use. A study was made of the effect of the different stages in the cold osmotic shock procedure, including the osmotic stress, the chelating agent, and the cold Mg(2+)-containing diluent, on viability and solute-accumulating ability. Growth of shocked cells in defined medium resembled that of unshocked cells; however, in malt extract-yeast extract-glucose-peptone medium, the shocked cells had a longer lag phase of growth and initially grew at a slower rate. Cold osmotic shock caused the release of low-molecular-weight compounds and about 6 to 8% of the cell protein. Neither the cell envelope enzymes, invertase, acid phosphatase and l-leucine-beta-naphthylamidase, nor the cytoplasmic enzyme, alkaline phosphatase, were released when yeast cells were subjected to cold osmotic shock.
酿酒酵母NCYC 366对冷渗透休克敏感。在30℃下于限定培养基中分批培养的指数生长期细胞，悬浮于含10 mM乙二胺四乙酸的0.8 m甘露醇中，然后再悬浮于冰冷的0.5 mM MgCl₂中后，与未受休克的细胞相比，积累非代谢性溶质d - 葡萄糖胺盐酸盐和2 - 氨基异丁酸的速率较慢；受休克的细胞保持了它们的活力。将未受休克的分批培养细胞在缓冲液中于10℃保存导致积累葡萄糖胺的能力增加，并且进一步的实验局限于在恒化器中在葡萄糖限制条件下生长的细胞，从而避免了在使用前储存细胞的需要。研究了冷渗透休克程序中不同阶段，包括渗透应激、螯合剂和含冷Mg²⁺的稀释剂，对活力和溶质积累能力的影响。在限定培养基中受休克细胞的生长类似于未受休克的细胞；然而，在麦芽提取物 - 酵母提取物 - 葡萄糖 - 蛋白胨培养基中，受休克的细胞有更长的生长延迟期并且最初生长速率较慢。冷渗透休克导致低分子量化合物和大约6%至8%的细胞蛋白释放。当酵母细胞受到冷渗透休克时，细胞包膜酶、转化酶、酸性磷酸酶和L - 亮氨酸 - β - 萘胺酶以及细胞质酶碱性磷酸酶均未释放。
