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Degradation of ribonucleic acid in Candida lipolytica : Extraction of ribonucleic acid degrading enzymes
Author(s) -
Imada Akira,
Sinskey Anthony J.,
Tannenbaum Steven R.
Publication year - 1972
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260140110
Subject(s) - rnase p , acetone , extraction (chemistry) , ribonuclease , chromatography , yield (engineering) , chemistry , enzyme , biochemistry , degradation (telecommunications) , ethanol , rna , telecommunications , materials science , computer science , metallurgy , gene
We developed an efficient and simple method for RNase extraction from Candida lipolytica cells which consists of predrying the cells with solvents and incubating them for 8 to 15 hr at 37 to 45°C in a slightly acid buffer which contains EDTA or salts. This method is called Solvent Dehydration Buffer Extraction (SDBE) procedure. Predrying with acetone or ethanol, or by lyophilization, followed by washing with acetone or ethylacetate gives the most efficient RNase extraction. The yield and specific activity obtained by this extraction procedure are higher than by any other method examined. An apparent 1.5‐ to 2.0‐fold activation of RNase occurred during the SDBE process. Activation of RNase in homogenates obtained by grinding fresh cells is also observed with EDTA or acetate buffer. The SDBE procedure works efficiently regardless of growth phase for Candida lipolytica , and works also with other Candida yeasts.