Purification of Rubisco from Leaves

Methods Mol Biol. 2024:2790:417-426. doi: 10.1007/978-1-0716-3790-6_22.


Rubisco fixes CO2 through the carboxylation of ribulose 1,5-bisphosphate (RuBP) during photosynthesis, enabling the synthesis of organic compounds. The natural diversity of Rubisco properties represents an opportunity to improve its performance and there is considerable research effort focusing on better understanding the properties and regulation of the enzyme. This chapter describes a method for large-scale purification of Rubisco from leaves. After the extraction of Rubisco from plant leaves, the enzyme is separated from other proteins by fractional precipitation with polyethylene glycol followed by ion-exchange chromatography. This method enables the isolation of Rubisco in large quantities for a wide range of biochemical applications.

Keywords: CO2 fixation; Calvin-Benson-Bassham cycle; Chromatography; Enzyme catalysis; Protein extraction; Ribulose-1,5-bisphosphate carboxylase/oxygenase.

MeSH terms

  • Chromatography, Ion Exchange / methods
  • Plant Leaves* / chemistry
  • Plant Leaves* / enzymology
  • Polyethylene Glycols / chemistry
  • Ribulose-Bisphosphate Carboxylase* / chemistry
  • Ribulose-Bisphosphate Carboxylase* / isolation & purification
  • Ribulose-Bisphosphate Carboxylase* / metabolism


  • Ribulose-Bisphosphate Carboxylase
  • Polyethylene Glycols