Cloning and expression in<i>Escherichia coli</i>of a phoA gene encoding a phosphate-irrepressible alkaline phosphatase of<i>Zymomonas mobilis</i>

Michel, G.; Alvarez, Elisa; Guzzo, Jean; Cami, B.; Baratti, J.

doi:10.1111/j.1574-6968.1992.tb05497.x

Cited by 4 publications

(2 citation statements)

References 13 publications

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“…However, from the information available in the literature, it is not clear whether the APase from 2. mobilis CP4 (database accession number L36230; P. F. Gomez & L. 0. Ingram, unpublished, 1994), which shows 34% identity to the Synechococcus PCC 7942 PhoV, is identical with the APase isolated and characterized from 2. mobilis ZM1 (Michel & Baratti, 1989;Michel et al, 1992;Baoudene-Assali et al, 1993).…”

Section: Mssdamssdnrhepaatlhtpahpvrivtaaslfdghdaainimrrll~qg~vihlgh Nmentioning

confidence: 99%

The cyanobacterium Synechococcus sp. strain PCC 7942 contains a second alkaline phosphatase encoded by phoV

1995

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A gene (phov) encoding an alkaline phosphatase from Synechococcus sp. strain PCC 7942 was isolated by screening a plasmid gene bank for expression of alkaline phosphatase activity in Escherichia coli JM103. Two independent clones carrying the same alkaline-phosphatase-encoding gene were isolated. One of these clones (pKW1) was further analysed and the nucleotide sequence of a contiguous 3234 bp DNA fragment was determined. Two complete open reading frames (ORFI and phoV) and an incomplete ORF3 were identified reading in the same direction. The deduced phoV gene product showed 34% identity to the alkaline phosphatase PhoA from Zymomonss mobilis, and the N-terminal part of the putative ORF3 protein exhibited 57% identity to a protein of unknown function from Frankia sp. Insertional inactivation of the Synechococcus PCC 7942 phoV gene failed, indicating an essential role for either the phoV or the ORF3 gene product. PhoV consists of 550 amino acid residues, resulting in a molecular mass of 61.3 kDa. To overexpress the Synechococcus PCC 7942 phoV gene in E. coli, plasmid pKWl was transformed into a phoA mutant of E. coli (CC118). In E. coli strain CC118(pKWI) PhoV was expressed constitutively with high rates of activity, and was shown to be membrane associated in the periplasmic space. After partial purification of the recombinant PhoV, it was shown that, like other alkaline phosphatases, the Synechococcus PhoV had a broad pH optimum in the alkaline region and a broad substrate specificity for phosphomonoesters, required Zn2+ for activity, and was inhibited by phosphate. In contrast to several other alkaline phosphatases, PhoV was inhibited by Mn2+. Due to the lack of a Synechococcus PCC 7942 phoV mutant strain, the function of PhoV remains uncertain. However, the present results show that Synechococcus PCC 7942 has a second, probably phosphate-irrepressible, alkaline phosphatase (PhoV, 61-3 kDa) in addition to the phosphate-repressible enzyme (PhoA, 145 kDa) already described.

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Section: Mssdamssdnrhepaatlhtpahpvrivtaaslfdghdaainimrrll~qg~vihlgh Nmentioning

confidence: 99%

The cyanobacterium Synechococcus sp. strain PCC 7942 contains a second alkaline phosphatase encoded by phoV

1995

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“…Most of these microbes carry the phosphatase (Pho) regulon containing coregulated genes involved in the synthesis of phosphatases, phosphate transporters and phosphonate utilization. Bacterial AlP production was reported to be coded as part of the Pho regulon by one of three homologue gene families (phoA, phoD, phoX) [11][12][13][14]. Metagenomic studies revealed that 32% of sequenced prokaryotic genomes contain at least one of the phoA, phoD or phoX genes, among which phoD is the most abundant bacterial AlP gene [15].…”

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confidence: 99%

Metagenomic Analysis of Bacterial Communities in Agricultural Soils from Vietnam with Special Attention to Phosphate Solubilizing Bacteria

2021

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Bacterial communities can promote increased phosphorus (P) availability for plants and microbes in soil via various mechanisms of phosphate solubilization. The production of extracellular phosphatases releases available P through the hydrolysis of organic P. Examining the abundance and diversity of the bacterial community, including phosphate solubilizing bacteria in soil, may provide valuable information to overcome P scarcity in soil ecosystems. Here, the diversity and relative abundance of bacterial phyla and genera of six agricultural soil samples from Vietnam were analysed by next generation sequencing of the 16S rRNA gene. Phosphatase activities of each soil were compared with physico-chemical parameters and the abundance of the alkaline phosphatase gene phoD. We showed the dominance of Chloroflexi, Proteobacteria, Actinobacteria, Acidobacteria and Firmicutes. Total nitrogen positively correlated with phyla Proteobacteria, Acidobacteria, Firmicutes and Planctomycetes. The abundance of several genera of Proteobacteria showed positive relationship with the copy number of the phoD gene. The abundance of several taxa positively correlated with silt content, while a negative relationship of Proteobacteria was found with sand content. Our results demonstrated the clear influence of soil physico-chemical properties on the abundance of various bacterial taxa including those potentially involved in phosphate solubilization.

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Cloning, sequencing and characterization of the alkaline phosphatase gene (phoD) fromZymomonas mobilis

Gomez

Ingram

1995

FEMS Microbiology Letters

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The phoD gene encoding the membrane-bound alkaline phosphatase (ALPI) from Zymomonas mobilis CP4 was cloned and sequenced. Both the translated sequence and the properties of the recombinant enzyme were unusual. Z. mobilis ALPI was monomeric (M(r) 62,926) and hydrolysed nucleotides more effectively than sugar phosphates. The translated sequence contained a single hydrophobic segment near the N-terminus which may serve as a membrane-anchor in Z. mobilis, although the recombinant enzyme was recovered in the cytoplasmic fraction of Escherichia coli. The predicted amino acid sequence for ALPI did not align well with other ALPs or other known genes. However, some similarity to E. coli ALP was noted in the metal-binding and phosphate-binding regions. Two other regions were identified with similarity to the active sites of pyruvate kinase and mammalian 5'-nucleotide phosphodiesterase (also membrane-bound), respectively. It is likely that Z. mobilis phoD represents a new class of alkaline phosphatase genes which has not been described previously.

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Cloning and expression inEscherichia coliof a phoA gene encoding a phosphate-irrepressible alkaline phosphatase ofZymomonas mobilis

Cited by 4 publications

References 13 publications

The cyanobacterium Synechococcus sp. strain PCC 7942 contains a second alkaline phosphatase encoded by phoV

The cyanobacterium Synechococcus sp. strain PCC 7942 contains a second alkaline phosphatase encoded by phoV

Metagenomic Analysis of Bacterial Communities in Agricultural Soils from Vietnam with Special Attention to Phosphate Solubilizing Bacteria

Cloning, sequencing and characterization of the alkaline phosphatase gene (phoD) fromZymomonas mobilis

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