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024 7 |a 10.1016/j.plantsci.2018.12.012  |2 doi 
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041 |a eng 
100 1 |a Muchut, Robertino J  |e verfasserin  |4 aut 
245 1 0 |a Heterologous expression and kinetic characterization of the α, β and αβ blend of the PPi-dependent phosphofructokinase from Citrus sinensis 
264 1 |c 2019 
336 |a Text  |b txt  |2 rdacontent 
337 |a ƒaComputermedien  |b c  |2 rdamedia 
338 |a ƒa Online-Ressource  |b cr  |2 rdacarrier 
500 |a Date Completed 18.03.2019 
500 |a Date Revised 30.09.2020 
500 |a published: Print-Electronic 
500 |a Citation Status MEDLINE 
520 |a Copyright © 2018 Elsevier B.V. All rights reserved. 
520 |a This work reports the molecular cloning and heterologous expression of the genes coding for α and β subunits of pyrophosphate-dependent phosphofructokinase (PPi-PFK) from orange. When expressed individually, both recombinant subunits were produced as highly purified monomeric proteins able to phosphorylate fructose-6-phosphate at the expenses of PPi (specific activity of 0.075 and 0.017 units. mg-1 for α and β subunits, respectively). On the other hand, co-expression rendered a α3β3 hexamer with specific activity three orders of magnitude higher than the single subunits. All the conformations of the enzyme were characterized with respect to its kinetic properties and sensitivity to the regulator fructose-2,6-bisphosphate. A thorough review of current knowledge on the matter indicates that this is the first report of the recombinant production of active plant PPi-PFK and the characterization of its different conformations. This is a main contribution for future studies focused to better understand the enzyme properties and how it accomplishes its relevant role in plant metabolism 
650 4 |a Journal Article 
650 4 |a Citrus sinensis 
650 4 |a Fructose-2,6-bisphosphate 
650 4 |a Hetero-hexamer 
650 4 |a Orange 
650 4 |a PPi-dependent phosphofructokinase 
650 4 |a Recombinant protein 
650 7 |a Diphosphates  |2 NLM 
650 7 |a Fructosediphosphates  |2 NLM 
650 7 |a Fructosephosphates  |2 NLM 
650 7 |a Multiprotein Complexes  |2 NLM 
650 7 |a Plant Proteins  |2 NLM 
650 7 |a Recombinant Proteins  |2 NLM 
650 7 |a diphosphoric acid  |2 NLM 
650 7 |a 4E862E7GRQ  |2 NLM 
650 7 |a fructose-6-phosphate  |2 NLM 
650 7 |a 6814-87-5  |2 NLM 
650 7 |a fructose 2,6-diphosphate  |2 NLM 
650 7 |a 79082-92-1  |2 NLM 
650 7 |a Phosphotransferases  |2 NLM 
650 7 |a EC 2.7.-  |2 NLM 
650 7 |a Phosphofructokinases  |2 NLM 
650 7 |a EC 2.7.1 -  |2 NLM 
650 7 |a pyrophosphate-fructose 6-phosphate 1-phosphotransferase  |2 NLM 
650 7 |a EC 2.7.1.90  |2 NLM 
700 1 |a Piattoni, Claudia V  |e verfasserin  |4 aut 
700 1 |a Margarit, Ezequiel  |e verfasserin  |4 aut 
700 1 |a Tripodi, Karina E J  |e verfasserin  |4 aut 
700 1 |a Podestá, Florencio E  |e verfasserin  |4 aut 
700 1 |a Iglesias, Alberto A  |e verfasserin  |4 aut 
773 0 8 |i Enthalten in  |t Plant science : an international journal of experimental plant biology  |d 1985  |g 280(2019) vom: 01. März, Seite 348-354  |w (DE-627)NLM098174193  |x 1873-2259  |7 nnns 
773 1 8 |g volume:280  |g year:2019  |g day:01  |g month:03  |g pages:348-354 
856 4 0 |u http://dx.doi.org/10.1016/j.plantsci.2018.12.012  |3 Volltext 
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952 |d 280  |j 2019  |b 01  |c 03  |h 348-354