Identification and characterization of alkaline phosphatase gene phoX in Microcystis aeruginosa PCC7806

Hong, Sujuan; Pan, Qianhui; Chen, Siyu; Zu, Yao; Xu, Chongxin; Li, Jianhong

doi:10.1007/s13205-021-02774-z

Cited by 10 publications

(4 citation statements)

References 38 publications

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“…This could also explain the decrease in the extracellular APase level in cell cultures with an OP source in this study, as OP was continuously transformed by APase, leading to the accumulation of PO 4 3– . Previous studies similarly showed an increase in the extracellular APase, followed by a decreased or constant APase content in Microcystis when using several OP sources (e.g., adenosine triphosphate and β-glycerophosphate), which is due to a continuous OP consumption rather than the inhibition of the related gene expression by PO 4 . − ,− ,,, …”

Section: Resultsmentioning

confidence: 76%

“…Previous studies similarly showed an increase in the extracellular APase, followed by a decreased or constant APase content in Microcystis when using several OP sources (e.g., adenosine triphosphate and β-glycerophosphate), which is due to a continuous OP consumption rather than the inhibition of the related gene expression by PO 4 . [3][4][5][7][8][9][10]14,60,61 The changes in TP and concentrations of extracellular APase attributed to Brucella sp. were relatively smaller than those attributed to Microcystis in the cocultures (Figures S4 and S5), indicating that the majority of P sources were converted and utilized by Microcystis, with a minor contribution from Brucella sp.…”

Section: Effects Of Brucella Sp On the Solubilization Of Limited Solu...mentioning

confidence: 99%

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Roles of Attached Bacteria on the Growth and Metabolism of Microcystis aeruginosa with Limited Soluble Phosphorus

Song,

Li,

Zou

et al. 2024

ACS EST Water

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The limited endogenous soluble phosphorus (P) in lake sediment is a potential and non-negligible P pool for the growth of Microcystis species and the bacteria attached to them. The attached bacteria are considered as P utilization supporters that contribute to Microcystis blooms; however, their effects on Microcystis growth and metabolism during the utilization of limited soluble P are poorly understood. Here, a series of cocultivations of Microcystis aeruginosa and a Microcystis growth-promoting bacterium, Brucella sp., were conducted using two P sources with limited solubility (tricalcium phosphate and calcium phytate). The results indicated that Brucella sp. secreted auxins to directly promote the growth of M. aeruginosa, and the secreted organic acids and alkaline phosphatase accelerated the overall P solubilization and transformation. In addition, the coexistence of M. aeruginosa with Brucella sp., which resulted in competition for P, stimulated M. aeruginosa to synthesize more photosynthetic pigments and alkaline phosphatase. Therefore, the results revealed that the attached bacteria improved the properties of M. aeruginosa and the phycosphere (e.g., the availability of P sources), providing comprehensive insights into their contribution to endogenous P utilization by Microcystis and the further occurrence of cyanobacterial blooms.

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Section: Resultsmentioning

confidence: 76%

Section: Effects Of Brucella Sp On the Solubilization Of Limited Solu...mentioning

confidence: 99%

Roles of Attached Bacteria on the Growth and Metabolism of Microcystis aeruginosa with Limited Soluble Phosphorus

Song,

Li,

Zou

et al. 2024

ACS EST Water

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“…The PhoX family phosphatases represent more realistic candidates for the release of phosphate in nature, however, these enzymes are not commercially available. PhoX APs are ubiquitous in marine microbes 28 , 33 , 37 – 39 secreted into the water 40 , operate in a wide range of environmental conditions 41 and are less restricted to a specific substrate 33 . Future studies will test PhoX in similar experiments.…”

Section: Discussionmentioning

confidence: 99%

Enzymatic phosphatization of fish scales—a pathway for fish fossilization

Gäb,

Bierbaum,

Wirth

et al. 2024

Sci Rep

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Phosphatized fish fossils occur in various locations worldwide. Although these fossils have been intensively studied over the past decades they remain a matter of ongoing research. The mechanism of the permineralization reaction itself remains still debated in the community. The mineralization in apatite of a whole fish requires a substantial amount of phosphate which is scarce in seawater, so the origin of the excess is unknown. Previous research has shown that alkaline phosphatase, a ubiquitous enzyme, can increase the phosphate content in vitro in a medium to the degree of saturation concerning apatite. We applied this principle to an experimental setup where fish scales were exposed to commercial bovine alkaline phosphatase. We analyzed the samples with SEM and TEM and found that apatite crystals had formed on the remaining soft tissue. A comparison of these newly formed apatite crystals with fish fossils from the Solnhofen and Santana fossil deposits showed striking similarities. Both are made up of almost identically sized and shaped nano-apatites. This suggests a common formation process: the spontaneous precipitation from an oversaturated solution. The excess activity of alkaline phosphatase could explain that effect. Therefore, our findings could provide insight into the formation of well-preserved fossils.

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“…PhoX of PCC7806 used as an antigen was generated as described by Hong et al (2021). The puri ed protein was checked by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and quanti ed by the Bradford assay (Bradford 1976).…”

Section: Expression and Puri Cation Of Phoxmentioning

confidence: 99%

Screening of nanobody targeting alkaline phosphatase PhoX in Microcystis and detection of the PhoX in situ by fluorescence immunoassays

Hong

Yin

et al. 2022

J Appl Phycol

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Cyanobacterial alkaline phosphatases (APases) play a key role in organophosphate utilization in freshwater. Tracking the distribution of APases could provide insights into the physiological response of phytoplankton to phosphorus nutrition. Extracellular APase PhoX, one of three prokaryotic APase families, is important for organophosphate utilization. Because the existing methods only give information on bulk APases activity, the speci c contribution of the PhoX is hardly evaluated. To develop an immunoassay for PhoX detection, the phoX gene of Microcystis aeruginosa PCC7806 was expressed in E. coli BL21 and the puri ed PhoX was used as a coated antigen for screening anti-PhoX nanobodies from an alpaca antibody library. After three rounds of panning, a nanobody 3H with the highest a nity was selected. Further tests showed 3H could speci cally bind to Microcystis PhoX and be used for PhoX detection by immunoblotting analysis. Then we constructed two different uorescent-labeled 3H, EGFP-3H (with an enhanced green uorescent protein, EGFP) and FITC-3H (with uorescein 5-isothiocyanate, FITC), as speci c uorescent dyes for PhoX. The uorescence staining tests with laboratory strains and water bloom samples showed that both uorescent-labeled nanobodies could visualize the distribution and evaluate relative expression levels of PhoX in Microcystis. The nanobody 3H could be a useful regent to develop uorescence immunoassays for in situ analyses of Microcystis PhoX.

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Identification and characterization of alkaline phosphatase gene phoX in Microcystis aeruginosa PCC7806

Cited by 10 publications

References 38 publications

Roles of Attached Bacteria on the Growth and Metabolism of Microcystis aeruginosa with Limited Soluble Phosphorus

Roles of Attached Bacteria on the Growth and Metabolism of Microcystis aeruginosa with Limited Soluble Phosphorus

Enzymatic phosphatization of fish scales—a pathway for fish fossilization

Screening of nanobody targeting alkaline phosphatase PhoX in Microcystis and detection of the PhoX in situ by fluorescence immunoassays

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