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用于降低产朊假丝酵母核酸含量的热休克过程的表征

Characterization of a heat-shock process for reduction of the nucleic acid content of Candida utilis.

作者信息

Ohta S, Maul S, Sinskey A J, Tannenbaum S R

出版信息

Appl Microbiol. 1971 Sep;22(3):415-21. doi: 10.1128/am.22.3.415-421.1971.

Abstract

A process for reducing the nucleic acid content of Candida utilis NRRL Y900 has been developed. The optimal process consists of heating the cells suspended in spent medium initially at pH 4.0 for various times at three different temperatures. Initially a heat-shock at 68 C for 1 to 3 sec is performed followed by incubation for 1 hr at 45 to 50 C and for a 2nd hr at 52 to 55 C. The distribution of degradation products has been characterized. Initially 90% of the nucleic acids were in a polymerized form (extractable by hot perchloric acid). After 30 min, much of this material was hydrolyzed but remained within the cell (extractable by cold perchloric acid). After 2 hr, most of the hydrolysis products leaked into the surrounding medium with only a small amount of low-molecular-weight material remaining within the membrane. Predominantly 3'-mononucleotides accumulated within the cell and eventually leaked from the cell.

摘要

已开发出一种降低产朊假丝酵母NRRL Y900核酸含量的方法。最佳方法包括将悬浮在废培养基中的细胞,最初在pH 4.0下于三种不同温度保温不同时间。首先在68℃进行1至3秒的热休克处理,随后在45至50℃孵育1小时,再在52至55℃孵育1小时。已对降解产物的分布进行了表征。最初,90%的核酸呈聚合形式(可被热高氯酸提取)。30分钟后,这种物质大部分被水解,但仍留在细胞内(可被冷高氯酸提取)。2小时后,大部分水解产物泄漏到周围培养基中,只有少量低分子量物质留在膜内。细胞内主要积累3'-单核苷酸,最终从细胞中泄漏出来。

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本文引用的文献

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Release of nucleotides from yeast cells.核苷酸从酵母细胞中的释放。
Nature. 1959 Oct 31;184:1381-3. doi: 10.1038/1841381a0.
7
PURIFICATION AND PROPERTIES OF RIBONUCLEASE FROM YEAST.酵母核糖核酸酶的纯化及性质
J Biochem. 1963 Oct;54:322-7. doi: 10.1093/oxfordjournals.jbchem.a127793.
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