A sensitive, simultaneous analysis of ribulose 1,5-bisphosphate carboxylase/oxygenase efficiencies: Graphical determination of the CO2/O 2 specificity factor.

Abstract:

:A simple approach to determine CO2/O2 specificity factor (τ) of ribulose 1,5-bisphosphate carboxylase/oxygenase is described. The assay measures the amount of CO2 fixation at varying [CO2]/[O2] ratios after complete consumption of ribulose 1,5-bisphosphate (RuBP). Carbon dioxide fixation catalyzed by the carboxylase was monitored by directly measuring the moles of (14)CO2 incorporated into 3-phosphoglycerate (PGA). This measurement at different [CO2]/[O2] ratios is used to determine graphically by several different linear plots the total RuBP consumed by the two activities and the CO2/O2 specificity factor. The assay can be used to measure the amounts of products of the carboxylase and oxygenase reactions and to determine the concentration of the substrate RuBP converted to an endpoint amount of PGA and phosphoglycolate. The assay was found to be suitable for all [CO2]/[O2] ratios examined, ranging from 14 to 215 micromolar CO2 (provided as 1-16 mM NaHCO3) and 614 micromolar O2 provided as 50% O2. The procedure described is extremely rapid and sensitive. Specificity factors for enzymes of highly divergent τ values are in good agreement with previously published data.

journal_name

Photosynth Res

journal_title

Photosynthesis research

authors

Kostov RV,McFadden BA

doi

10.1007/BF00029463

subject

Has Abstract

pub_date

1995-01-01 00:00:00

pages

57-66

issue

1

eissn

0166-8595

issn

1573-5079

journal_volume

43

pub_type

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