Abstract
Burrous, S. E. (University of Illinois, Urbana) and W. A. Wood. Enzyme distribution in membrane, cytoplasm, and nuclear fractions from Pseudomonas fluorescens. J. Bacteriol. 84:364–369. 1962.—A procedure is described for fractionation of Pseudomonas fluorescens A3.12 by controlled rupture of spheroplasts and fractional centrifugations to yield cytoplasm, membrane, and deoxyribonucleic acid (DNA) fractions with low cross contamination and little loss of enzymatic activity. Assays for the content of a number of oxidative and nonoxidative enzymes revealed that: (i) little activity is present in the DNA fraction; (ii) only reduced diphosphopyridine nucleotide oxidase is found exclusively in the membranes; (iii) phosphohexosisomerase and probably transketolase and glucose-6-phosphate dehydrogenase are found exclusively in the cytoplasm; and (iv) the remaining five enzymes studied were found in both fractions. In contrast to earlier reports of gluconic dehydrogenase being membrane-bound and 2-keto-3-deoxy-6-phosphogluconate aldolase being soluble in sonic extracts, this procedure produced partly soluble gluconic dehydrogenase and partly membrane-associated 2-keto-3-deoxy-6-phosphogluconate aldolase.
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
- FROMM H. J. Ribitol dehydrogenase. I. Purification and properties of the enzyme. J Biol Chem. 1958 Nov;233(5):1049–1052. [PubMed] [Google Scholar]
- KOVACHEVICH R., WOOD W. A. Carbohydrate metabolism by Pseudomonas fluorescens. IV. Purification and properties of 2-keto-3-deoxy-6-phosphogluconate aldolase. J Biol Chem. 1955 Apr;213(2):757–767. [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]
- OGUR M., ROSEN G. The nucleic acids of plant tissues; the extraction and estimation of desoxypentose nucleic acid and pentose nucleic acid. Arch Biochem. 1950 Feb;25(2):262–276. [PubMed] [Google Scholar]
- REPASKE R. Lysis of gram-negative bacteria by lysozyme. Biochim Biophys Acta. 1956 Oct;22(1):189–191. doi: 10.1016/0006-3002(56)90240-2. [DOI] [PubMed] [Google Scholar]
- ROBRISH S. A., MARR A. G. Location of enzymes in Azotobacteragilis. J Bacteriol. 1962 Jan;83:158–168. doi: 10.1128/jb.83.1.158-168.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SIMPSON F. J., WOLIN M. J., WOOD W. A. Degradation of L-arabinose by Aerobacter aerogenes. I. A pathway involving phosphorylated intermediates. J Biol Chem. 1958 Jan;230(1):457–472. [PubMed] [Google Scholar]
- SPIEGELMAN S., ARONSON A. I., FITZJAMES P. C. Isolation and characterization of nuclear bodies from protoplasts of Bacillus megaterium. J Bacteriol. 1958 Jan;75(1):102–117. doi: 10.1128/jb.75.1.102-117.1958. [DOI] [PMC free article] [PubMed] [Google Scholar]
- WEIBULL C. Characterization of the protoplasmic constituents of bacillus megaterium. J Bacteriol. 1953 Dec;66(6):696–702. doi: 10.1128/jb.66.6.696-702.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]
- WEIBULL C. The isolation of protoplasts from Bacillus megaterium by controlled treatment with lysozyme. J Bacteriol. 1953 Dec;66(6):688–695. doi: 10.1128/jb.66.6.688-695.1953. [DOI] [PMC free article] [PubMed] [Google Scholar]
- WOOD W. A., SCHWERDT R. F. Carbohydrate oxidation by Pseudomonas fluorescens. II. Mechanism of hexose phosphate oxidation. J Biol Chem. 1954 Feb;206(2):625–635. [PubMed] [Google Scholar]