Identification and characterization of ChlreSEX4, a novel glucan phosphatase from Chlamydomonas reinhardtii green alga.,Archives of Biochemistry and Biophysics

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Identification and characterization of ChlreSEX4, a novel glucan phosphatase from Chlamydomonas reinhardtii green alga.
Archives of Biochemistry and Biophysics ( IF 3.8 ) Pub Date : 2019-12-23 , DOI: 10.1016/j.abb.2019.108235
Julieta B Carrillo ₁ , Florencia Torresi ₁ , Luisina L Morales ₁ , Micaela Ricordi ₁ , Diego F Gomez-Casati ₁ , Maria V Busi ₁ , Mariana Martín ₁

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Chlamydomonas reinhardtii is the best known unicellular green alga model which has long been used to investigate all kinds of cellular processes, including starch metabolism. Here we identified and characterized a novel enzyme, ChlreSEX4, orthologous to glucan phosphatase SEX4 from Arabidopsis thaliana, that is capable of binding and dephosphorylating amylopectin in vitro. We also reported that cysteine 224 and tryptophan 305 residues are critical for enzyme catalysis and substrate binding. Furthermore, we verified that ChlreSEX4 gene is expressed in vivo and that glucan phosphatase activity is measurable in Chlamydomonas protein extracts. In view of the results presented, we suggest ChlreSEX4 as a functional phosphoglucan phosphatase from C. reinhardtii. Our data obtained so far contribute to understanding the phosphoglucan phosphatases evolutionary process in the green lineage and their role in starch reversible phosphorylation. In addition, this allows to position Chlamydomonas as a potential tool to obtain starches with different degrees of phosphorylation for industrial or biotechnological purposes.

中文翻译：

ChlreSEX4的鉴定和表征，一种来自莱茵衣藻的新型葡聚糖磷酸酶。

莱茵衣藻是最著名的单细胞绿藻模型，长期以来一直用于研究各种细胞过程，包括淀粉代谢。在这里，我们鉴定并鉴定了一种新型酶ChlreSEX4，它与拟南芥葡聚糖磷酸酶SEX4同源，能够在体外结合和脱支支链淀粉。我们还报道了半胱氨酸224和色氨酸305残基对于酶催化和底物结合至关重要。此外，我们验证了ChlreSEX4基因在体内表达，并且在衣藻蛋白提取物中可检测到葡聚糖磷酸酶活性。鉴于呈现的结果，我们建议ChlreSEX4作为来自莱茵衣藻的功能性磷酸葡聚糖磷酸酶。到目前为止，我们获得的数据有助于理解绿色谱系中的磷酸葡聚糖磷酸酶的进化过程及其在淀粉可逆磷酸化中的作用。另外，这允许将衣藻定位为用于工业或生物技术目的获得具有不同磷酸化程度的淀粉的潜在工具。

更新日期：2019-12-23

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