Preiser Alyssa L, Banerjee Aparajita, Weise Sean E, Renna Luciana, Brandizzi Federica, Sharkey Thomas D
MSU-DOE Plant Research Laboratory, Michigan State University, East Lansing, MI, United States.
Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI, United States.
Front Plant Sci. 2020 Dec 10;11:580726. doi: 10.3389/fpls.2020.580726. eCollection 2020.
Phosphoglucoisomerase (PGI) isomerizes fructose 6-phosphate (F6P) and glucose 6-phosphate (G6P) in starch and sucrose biosynthesis. Both plastidic and cytosolic isoforms are found in plant leaves. Using recombinant enzymes and isolated chloroplasts, we have characterized the plastidic and cytosolic isoforms of PGI. We have found that the plastidic PGI for G6P is three-fold greater compared to that for F6P and that erythrose 4-phosphate is a key regulator of PGI activity. Additionally, the of spinach plastidic PGI can be dynamically regulated in the dark compared to the light and increases by 200% in the dark. We also found that targeting cytosolic PGI into plastids of disrupts starch accumulation and degradation. Our results, in combination with the observation that plastidic PGI is not in equilibrium, indicates that PGI is an important regulatory enzyme that restricts flow and acts as a one-way valve preventing backflow of G6P into the Calvin-Benson cycle. We propose the PGI may be manipulated to improve flow of carbon to desired targets of biotechnology.
磷酸葡萄糖异构酶(PGI)在淀粉和蔗糖生物合成过程中催化果糖6-磷酸(F6P)和葡萄糖6-磷酸(G6P)的异构化反应。植物叶片中同时存在质体和胞质两种同工型。我们利用重组酶和分离的叶绿体对PGI的质体和胞质同工型进行了表征。我们发现,质体PGI对G6P的亲和力比对F6P高3倍,并且4-磷酸赤藓糖是PGI活性的关键调节因子。此外,与光照条件相比,菠菜质体PGI在黑暗中可被动态调节,活性在黑暗中增加200%。我们还发现,将胞质PGI靶向导入缺乏该酶的植物质体中会扰乱淀粉的积累和降解。我们的研究结果,结合质体PGI并非处于平衡状态这一观察结果,表明PGI是一种重要的调节酶,它限制了碳流,并作为单向阀防止G6P回流到卡尔文-本森循环中。我们提出,可以通过调控PGI来改善碳流向生物技术所需目标的流量。