Tsuji K
Appl Microbiol. 1966 May;14(3):456-61. doi: 10.1128/am.14.3.456-461.1966.
Growth medium as well as freezing menstruum greatly influenced the recovery of Saccharomyces carlsbergensis when it was quickly frozen in liquid nitrogen at - 196 C and quickly thawed at 40 C. Nearly 90% recovery in viability was obtained when S. carlsbergensis was grown in Trypticase Soy Broth and frozen in vitamin B(6) basal assay medium. The growth phase of S. carlsbergensis also influenced recovery after freezing. When S. carlsbergensis was grown in Trypticase Soy Broth and frozen in the broth at the logarithmic-growth phase, only 7% viability was retained; the recovery rate increased to 81% when the culture was frozen in the maximal stationary phase. To have the least possible lag period of growth after thawing, a technique called growth-phase conditioning was introduced. After 1 hr of growth-phase conditioning, S. carlsbergensis was clearly out of lag phase, and budding was observed. A vitamin B(6) microbiological assay with a 6-hr incubation period and with the use of liquid nitrogen-frozen S. carlsbergensis is described.
当在-196℃的液氮中快速冷冻并在40℃快速解冻时，生长培养基以及冷冻介质对卡尔斯伯酵母的复苏有很大影响。当卡尔斯伯酵母在胰蛋白胨大豆肉汤中生长并在维生素B6基础测定培养基中冷冻时，可获得近90%的活力恢复。卡尔斯伯酵母的生长阶段也会影响冷冻后的复苏情况。当卡尔斯伯酵母在胰蛋白胨大豆肉汤中生长并在对数生长期的肉汤中冷冻时，仅保留7%的活力；当培养物在最大稳定期冷冻时，复苏率提高到81%。为了使解冻后的生长滞后期尽可能短，引入了一种称为生长阶段调节的技术。经过1小时的生长阶段调节后，卡尔斯伯酵母明显脱离滞后期，并观察到出芽现象。本文描述了一种使用液氮冷冻的卡尔斯伯酵母、培养6小时的维生素B6微生物测定法。
