Hong Sujuan, Pan Qianhui, Chen Siyu, Zu Yao, Xu Chongxin, Li Jianhong
School of Life Sciences, Nanjing Normal University, Nanjing, 210023 China.
Laboratory for Food Quality, Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology, Institute of Food Safety and Nutrition, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014 China.
3 Biotech. 2021 May;11(5):218. doi: 10.1007/s13205-021-02774-z. Epub 2021 Apr 16.
PhoX is an extracellular alkaline phosphatase that is widely found in cyanobacteria and plays an important role in the conversion of extracellular organophosphorus into soluble inorganic phosphorus. However, the gene has not yet been experimentally confirmed to exist in bloom-forming species. In this study, we identified a putative gene (GenBank accession no. ARI79942.1) in PCC7806 and overexpressed it in 21 (DE3). The expressed PhoX protein displayed phosphodiesterase and phosphomonoesterase activities. In contrast to other bacterial PhoX proteins, which are activated mainly by Ca, PhoX was most strongly activated by Mg, followed by Co, Ca, Zn and Mn, but it was inhibited by Ni. Sequence analysis showed that was highly conserved in the genus (DNA similarity > 96% between species). expression responded significantly to different environmental phosphorus levels. When PCC7806 cells were cultured in phosphorus-deficient medium (BG11-P), expression reached its highest level at 2 h and then decreased to a low level at 4 h. Organophosphate induced the expression of ; its expression reached the highest level at 4 h and was maintained at a high level at 6 h. Our results confirmed a putative gene and demonstrated that the gene of is conserved.
PhoX是一种细胞外碱性磷酸酶,广泛存在于蓝藻中,在将细胞外有机磷转化为可溶性无机磷的过程中发挥重要作用。然而,该基因尚未在形成水华的物种中通过实验得到证实。在本研究中,我们在集胞藻PCC7806中鉴定出一个假定基因(GenBank登录号:ARI79942.1),并在大肠杆菌BL21(DE3)中对其进行了过表达。表达的PhoX蛋白表现出磷酸二酯酶和磷酸单酯酶活性。与其他主要由Ca激活的细菌PhoX蛋白不同,PhoX受Mg激活作用最强,其次是Co、Ca、Zn和Mn,但受Ni抑制。序列分析表明,该基因在集胞藻属中高度保守(物种间DNA相似性>96%)。PhoX的表达对不同的环境磷水平有显著响应。当集胞藻PCC7806细胞在缺磷培养基(BG11-P)中培养时,PhoX的表达在2小时达到最高水平,然后在4小时降至低水平。有机磷酸盐诱导PhoX的表达;其表达在4小时达到最高水平,并在6小时维持在高水平。我们的结果证实了一个假定的PhoX基因,并证明集胞藻的PhoX基因是保守的。