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. 1970 Apr;117(2):215–220. doi: 10.1042/bj1170215

Coproporphyrinogenase in tobacco (Nicotiana tabacum L.)

W P Hsu 1,*, G W Miller 1,
PMCID: PMC1178852  PMID: 4986869

Abstract

1. Coproporphyrinogenase was extracted and purified from tobacco (Nicotiana tabacum L.). Enzyme activity was mainly located in mitochondria rather than in chloroplasts. The enzyme was purified by differential centrifugation, ammonium sulphate fractionation, calcium phosphate gel adsorption and dialysis. A 69-fold final purification was obtained. 2. An apparent Km value of 3.6×10−5m was found, the value being largely dependent on the amount of coproporphyrin III recovered after reduction with sodium amalgam to coproporphyrinogen III. Protoporphyrin formation was linear up to 3h and decreased with further incubation. The enzyme activity increased with the concentration of enzyme protein up to 30μg/ml of solution. 3. Enzyme activity was greatly enhanced by increasing Fe2+ concentrations up to 0.5mm, beyond which inhibition occurred. Co2+ and Mn2+ were also found to activate at low concentrations (0.1mm) and inhibit at higher concentrations (5mm). Fe3+ and Cu2+, both at 0.1mm, and o-phenanthroline and EDTA, each at 1mm, were found to be inhibitory.

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Selected References

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  1. Adler A. D., George P. Determination of iron in heme compounds. I. Hemin. Anal Biochem. 1965 May;11(2):159–163. doi: 10.1016/0003-2697(65)90001-1. [DOI] [PubMed] [Google Scholar]
  2. Cameron B. F. Determination of iron in heme compounds. II. Hemoglobin and myoglobin. Anal Biochem. 1965 May;11(2):164–169. doi: 10.1016/0003-2697(65)90002-3. [DOI] [PubMed] [Google Scholar]
  3. FALK J. E. Chromatography of porphyrins and metalloporphyrins. J Chromatogr. 1961 Apr;5:277–299. doi: 10.1016/s0021-9673(01)92860-2. [DOI] [PubMed] [Google Scholar]
  4. Hsu W. P., Miller G. W. Chlorophyll and porphyrin synthesis in relation to iron in Nicotiana tabacum, L. Biochim Biophys Acta. 1965 Dec 16;111(2):393–402. doi: 10.1016/0304-4165(65)90049-8. [DOI] [PubMed] [Google Scholar]
  5. LASCELLES J. Synthesis of porphyrins by cell suspensions of Tetrahymena vorax: effect of members of the vitamin B group. Biochem J. 1957 May;66(1):65–72. doi: 10.1042/bj0660065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. LASCELLES J. The synthesis of porphyrins and bacteriochlorophyll by cell suspensions of Rhodopseudomonas spheroides. Biochem J. 1956 Jan;62(1):78–93. doi: 10.1042/bj0620078. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Marsh H. V., Evans H. J., Matrone G. Investigations of the Role of Iron in Chlorophyll Metabolism I. Effect of Iron Deficiency on Chlorophyll and Heme Content and on the Activities of Certain Enzymes in Leaves. Plant Physiol. 1963 Nov;38(6):632–638. doi: 10.1104/pp.38.6.632. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Marsh H. V., Evans H. J., Matrone G. Investigations of the Role of Iron in Chlorophyll Metabolism. II. Effect of Iron Deficiency on Chlorophyll Synthesis. Plant Physiol. 1963 Nov;38(6):638–642. doi: 10.1104/pp.38.6.638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. RIMINGTON C., SVEINSSON S. L. The spectrophotometric determination of uroporphyrin. Scand J Clin Lab Invest. 1950;2(3):209–216. doi: 10.3109/00365515009049872. [DOI] [PubMed] [Google Scholar]
  10. SANO S., GRANICK S. Mitochondrial coproporphyrinogen oxidase and protoporphyrin formation. J Biol Chem. 1961 Apr;236:1173–1180. [PubMed] [Google Scholar]
  11. del Batlle A. M., Benson A., Rimington C. Purification and properties of coproporphyrinogenase. Biochem J. 1965 Dec;97(3):731–740. doi: 10.1042/bj0970731. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. van Niel C. B. THE CULTURE, GENERAL PHYSIOLOGY, MORPHOLOGY, AND CLASSIFICATION OF THE NON-SULFUR PURPLE AND BROWN BACTERIA. Bacteriol Rev. 1944 Mar;8(1):1–118. doi: 10.1128/br.8.1.1-118.1944. [DOI] [PMC free article] [PubMed] [Google Scholar]

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