Abstract
A ribonuclease (RNAase; EC 3.1.14.1) from brewer's yeast was purified 90-fold. Crude RNAase was initially separated from other proteins by precipitation at pH 4.0 after incubation of the mechanically disrupted yeast cells at pH 6.0 and 52 degrees C for 30 min. The RNAase was purified from the supernatant by ultrafiltration with a PM-30 membrane and adsorption chromatography on hydroxyapatite. RNAase preparation was free of phosphatase, deoxyribonuclease and phosphodiesterase activities. It showed maximum activity at pH 6.0 and a temperature optimum of 52 degrees C with yeast RNA as substrate. This RNAase hydrolysed yeast RNA to nucleoside 3'-phosphates and showed no evidence of base specificity.
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- DISCHE Z., LANDSBERG E. A color reaction of pentose phosphate esters substituted in position 5. Biochim Biophys Acta. 1957 Apr;24(1):193–195. doi: 10.1016/0006-3002(57)90163-4. [DOI] [PubMed] [Google Scholar]
- FRISCH-NIGGEMEYER W., REDDI K. K. Studies on ribonuclease in tobacco leaves. I. Purification and properties. Biochim Biophys Acta. 1957 Oct;26(1):40–46. doi: 10.1016/0006-3002(57)90051-3. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Lindblom M. Properties of intracellular ribonuclease utilized for RNA reduction in disintegrated cells of Saccharomyces cerevisiae. Biotechnol Bioeng. 1977 Feb;19(2):199–210. doi: 10.1002/bit.260190204. [DOI] [PubMed] [Google Scholar]
- MARKHAM R., SMITH J. D. Structure of ribonucleic acid. Nature. 1951 Sep 8;168(4271):406–408. doi: 10.1038/168406a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
- OHTAKA Y., UCHIDA K., SAKAI T. PURIFICATION AND PROPERTIES OF RIBONUCLEASE FROM YEAST. J Biochem. 1963 Oct;54:322–327. doi: 10.1093/oxfordjournals.jbchem.a127793. [DOI] [PubMed] [Google Scholar]
- Pringle J. R. Methods for avoiding proteolytic artefacts in studies of enzymes and other proteins from yeasts. Methods Cell Biol. 1975;12:149–184. doi: 10.1016/s0091-679x(08)60956-5. [DOI] [PubMed] [Google Scholar]
- TAKAHASHI K. The structure and function of ribonuclease T1. II. Further purification and amino acid composition of ribonuclease T1. J Biochem. 1962 Feb;51:95–108. doi: 10.1093/oxfordjournals.jbchem.a127515. [DOI] [PubMed] [Google Scholar]