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. 1982 Oct;70(4):1213–1217. doi: 10.1104/pp.70.4.1213

Isolation and Purification of Intact Peroxisomes from Green Leaf Tissue 1

Mark R Schmitt 1,2,3,2, Gerald E Edwards 1,2,3
PMCID: PMC1065852  PMID: 16662640

Abstract

Intact peroxisomes were prepared from green leaves of a number of C3 species, both monocots and dicots. A protoplast extract from which chloroplasts have been removed by a 1-minute 10,000g centrifugation was applied to a step gradient of 5, 15, 30, and 45% Percoll containing 0.5 molar sucrose, 0.1% BSA, and 25 millimolar Hepes-KOH (pH 7.5). After centrifugation, a peroxisomal fraction with low contamination by chloroplastic and mitochondrial markers was recovered from the 30/45% Percoll interface. This fraction was passed through a Sepharose 2B column to remove the Percoll which resulted in a peroxisomal preparation exhibiting high intactness (estimated from enzyme latency) and stability.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arnon D. I. COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. Plant Physiol. 1949 Jan;24(1):1–15. doi: 10.1104/pp.24.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Hatch M. D. A simple spectrophotometric assay for fumarate hydratase in crude tissue extracts. Anal Biochem. 1978 Mar;85(1):271–275. doi: 10.1016/0003-2697(78)90299-3. [DOI] [PubMed] [Google Scholar]
  3. Kohn L. D., Warren W. A. The kinetic properties of spinach leaf glyoxylic acid reductase. J Biol Chem. 1970 Aug 10;245(15):3831–3839. [PubMed] [Google Scholar]
  4. Latzko E., Gibbs M. Enzyme activities of the carbon reduction cycle in some photosynthetic organisms. Plant Physiol. 1969 Feb;44(2):295–300. doi: 10.1104/pp.44.2.295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Nishimura M., Douce R., Akazawa T. Isolation and characterization of metabolically competent mitochondria from spinach leaf protoplasts. Plant Physiol. 1982 Apr;69(4):916–920. doi: 10.1104/pp.69.4.916. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Stitt M., Heldt H. W. Physiological rates of starch breakdown in isolated intact spinach chloroplasts. Plant Physiol. 1981 Sep;68(3):755–761. doi: 10.1104/pp.68.3.755. [DOI] [PMC free article] [PubMed] [Google Scholar]

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