Isolation of ribulose-1,5-bisphosphate carboxylase/oxygenase from leaves.
Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is a multifunctional enzyme that catalyzes the fixation of CO2 and O2 in photosynthesis and photorespiration, respectively. As the rate-limiting step in photosynthesis, improving the catalytic properties of Rubisco has long been viewed as a viable strategy for increasing plant productivity. Advances in biotechnology have made this goal more attainable by making it possible to modify Rubisco in planta. To properly evaluate the properties of Rubisco, it is necessary to isolate the enzyme in pure form. This chapter describes procedures for rapid and efficient purification of Rubisco from leaves of several species.
Carmo-Silva
A Elizabete
AE
U.S. Department of Agriculture, Arid-Land Agricultural Research Center, Maricopa, AZ, USA.
Barta
Csengele
C
Salvucci
Michael E
ME
D000645:Ammonium Sulfate / Q000737:chemistry; D002498:Centrifugation; D005591:Chemical Fractionation / Q000379:methods*; D011232:Chemical Precipitation; D002852:Chromatography, Ion Exchange; D018515:Plant Leaves / Q000201:enzymology*; D010944:Plants / Q000201:enzymology*; D012273:Ribulose-Bisphosphate Carboxylase / Q000737:chemistry / Q000302:isolation & purification*; D013395:Sucrose / Q000737:chemistry
D013395:Sucrose; D012273:Ribulose-Bisphosphate Carboxylase; D000645:Ammonium Sulfate