期刊
MICROORGANISMS
卷 7, 期 8, 页码 -出版社
MDPI
DOI: 10.3390/microorganisms7080232
关键词
dinoflagellate; alkaline phosphatase; metal cofactor; inducible expression
类别
资金
- National Key Research and Development Program of China [2017YFC1404302]
- National Natural Science Foundation of China [41776116, 31661143029, 41706116]
- Fundamental Research Funds for the Central Universities [20720180101]
Alkaline phosphatase (AP) enables marine phytoplankton to utilize dissolved organic phosphorus (DOP) when dissolved inorganic phosphate (DIP) is depleted in the ocean. Dinoflagellate AP (Dino-AP) represents a newly classified atypical type of AP, PhoA(aty). Despite While being a conventional AP, PhoA(EC) is known to recruit Zn2+ and Mg2+ in the active center, and the cofactors required by PhoA(aty) have been contended and remain unclear. In this study, we investigated the metal ion requirement of AP in five dinoflagellate species. After AP activity was eliminated by using EDTA to chelate metal ions, the enzymatic activity could be recovered by the supplementation of Ca2+, Mg2+ and Mn2+ in all cases but not by that of Zn2+. Furthermore, the same analysis conducted on the purified recombinant ACAAP (AP of Amphidinium carterae) verified that the enzyme could be activated by Ca2+, Mg2+, and Mn2+ but not Zn2+. We further developed an antiserum against ACAAP, and a western blot analysis using this antibody showed a remarkable up-regulation of ACAAP under a phosphate limitation, consistent with elevated AP activity. The unconventional metal cofactor requirement of Dino-AP may be an adaptation to trace metal limitations in the ocean, which warrants further research to understand the niche differentiation between dinoflagellates and other phytoplankton that use Zn-Mg AP in utilizing DOP.
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