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A Rapid and Sensitive Method for Determination of Relative Specificity of RuBisCO from Various Species by Anion-Exchange Chromatography |
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| Authors: | Uemura Koichi; Suzuki Yoshihiro; Shikanai Toshiharu; Wadano Akira; Jensen Richard G; Chmara Wendy; Yokota Akiho |
| Institution: | 1Plant Molecular Physiology Laboratory, Research Institute of Innovative Technology for the Earth (RITE) Kizu, Kyoto, 619-02 Japan 2Department of Agricultural Chemistry, University of Osaka Prefecture Sakai, Osaka, 591 Japan 3Department of Biochemistry and Plant Science, University of Arizona Tucson, Arizona 85721, U.S.A. |
| Abstract: | A rapid and sensitive method to determine the relative specificity( ) of ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO)using anion-exchange chromatography is described. We employedthe open gas system for the reaction of RuBisCO to get the mostreliable CO2 and O2 concentrations in the reaction mixture.3-Phosphoglycerate and 2-phosphoglycolate which were formedin the RuBisCO reaction were completely separated and directlymeasured with anion-exchange chromatography without using radioisotopes.The determination of the value was accomplished in 3 h. The values of RuBisCO enzymes from higher land plants were between90 and 96, and those from bacteria including cyanobacteriumwere close to 45. These values were in agreement with previouslyreported values. The enzyme of the red macroalga Porphyra yezoensisexhibited a value of over 140, as expected from the reportedvalue of the enzyme from the red microalga Porphyridium cruenteum.RuBisCO from the green macroalga Ulva pertusa had a value closeto 70 and similar to that of the enzyme from the green microalgaChlamydomonas reinhardtii.
4On leave from Research and Development Center, Unitika Ltd.,23 Kozakura, Uji, Kyoto, 611 Japan.
5Present address: Nara Institute of Science and Technology,8916-5 Takayama-Cho, Ikoma, Nara, 630-01 Japan |
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