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Date of Publication:2015-01-01

Journal:北京理工大学学报

Issue:9

Page Number:985-990

ISSN No.:1001-0645

Abstract:Starch, synthesized by microalgae with utilization of light energy and carbon dioxide, is the sustainable feedstock for large scale production of fuel ethanol. In order to enhance accumulation of starch by genetic engineering, it is essential to understand catalytic process and regulation of the key enzymes of starch metabolism in microalgae. For this reason, the rate-limiting enzyme of starch biosynthesis, ADP-glucose pyrophosphorylase (AGPase) in Chlamydomonas reinhardtii, was selected as bait protein for protein-protein interaction (PPI) investigation. With acquisition of AGPase-GST fusion protein, GST-pull down assay was introduced to detect the interaction connected with AGPase. The acquired proteins were further separated and analyzed qualitatively by HPLC-MS/MS. Subsequently, the data was mining by bioinformatics tools. Finally, eleven kinds of proteins were speculated to interact with AGPase, including molecular chaperones, photosynthesis related proteins and signaling molecules. The biological function behind the interaction between these proteins and AGPase can be inferred from the analysis of their subcellular locations and physiological feature. © 2015, Beijing Institute of Technology. All right reserved.

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