Identification of an Escherichia coli periplasmic acid phosphatase containing of a 27 kDa-polypeptide component

Rossolini, G. M.

doi:10.1016/0378-1097(94)90614-9

Cited by 19 publications

(50 citation statements)

References 12 publications

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“…Here we present evidence that these mutants actually lack the periplasmic acid phosphatase AphA (18,19,33,40,42), which removes phosphate from NMN in the periplasm and produces NmR for transport by PnuC. Evidence reported here supports reinterpretation of four previous conclusions regarding NMN assimilation in Salmonella.…”

supporting

confidence: 87%

Assimilation of Nicotinamide Mononucleotide Requires Periplasmic AphA Phosphatase inSalmonella enterica

Grose

Bergthorsson

et al. 2005

J Bacteriol

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Salmonella enterica can obtain pyridine from exogenous nicotinamide mononucleotide (NMN) by three routes. In route 1, nicotinamide is removed from NMN in the periplasm and enters the cell as the free base. In route 2, described here, phosphate is removed from NMN in the periplasm by acid phosphatase (AphA), and the produced nicotinamide ribonucleoside (NmR) enters the cell via the PnuC transporter. Internal NmR is then converted back to NMN by the NmR kinase activity of NadR. Route 3 is seen only in pnuC* transporter mutants, which import NMN intact and can therefore grow on lower levels of NMN. Internal NMN produced by either route 2 or route 3 is deamidated to nicotinic acid mononucleotide and converted to NAD by the biosynthetic enzymes NadD and NadE.

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supporting

confidence: 87%

Assimilation of Nicotinamide Mononucleotide Requires Periplasmic AphA Phosphatase inSalmonella enterica

Grose

Bergthorsson

et al. 2005

J Bacteriol

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“…Estimation of the recombinant enzyme's molecular mass in the denatured state was performed on a NuPAGE 4 to 12% Bis-Tris gel (Invitrogen) utilized per the manufacturer's recommendation. A modified version of the zymogram technique of Rossolini et al (40) following SDS-PAGE and renaturation was used to qualitatively assess the in situ phosphomonoesterase activity of the purification samples. 4-Methylumbelliferyl phosphate (4MUP) at a final concentration of 1 mM was used as the substrate in the zymogram assay buffer.…”

Section: Methodsmentioning

confidence: 99%

Characterization of a Unique Class C Acid Phosphatase from Clostridium perfringens

Reilly¹,

Chance²,

Calcutt³

et al. 2009

Appl Environ Microbiol

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Clostridium perfringens is a gram-positive anaerobe and a pathogen of medical importance. The detection of acid phosphatase activity is a powerful diagnostic indicator of the presence of C. perfringens among anaerobic isolates; however, characterization of the enzyme has not previously been reported. Provided here are details of the characterization of a soluble recombinant form of this cell-associated enzyme. The denatured enzyme was ϳ31 kDa and a homodimer in solution. It catalyzed the hydrolysis of several substrates, including para-nitrophenyl phosphate, 4-methylumbelliferyl phosphate, and 3 and 5 nucleoside monophosphates at pH 6. Calculated K m s ranged from 0.2 to 0.6 mM with maximum velocity ranging from 0.8 to 1.6 mol of P i /s/mg. Activity was enhanced in the presence of some divalent cations but diminished in the presence of others. Wild-type enzyme was detected in all clinical C. perfringens isolates tested and found to be cell associated. The described enzyme belongs to nonspecific acid phosphatase class C but is devoid of lipid modification commonly attributed to this class.Clostridium perfringens is a ubiquitous gram-positive, endospore-forming anaerobic bacterium. While found in soil, the organism is often a constituent of the intestinal flora and is capable of causing severe gastrointestinal and histotoxic infections in humans and animals. As a species, C. perfringens is a very heterogeneous group of organisms with respect to metabolic by-products produced in vitro and pathogenic potential (25). Although the literature is replete with descriptions of the immunological, bio-and physicochemical attributes of C. perfringens-encoded toxins, as well as molecular and morphological details of sporulation, detailed analysis of other constituent enzymes, including those involved in acquisition and regulation of inorganic phosphate, has received relatively little attention. This lack of information is notable. A growing cadre of investigators (1, 7) has recognized the clinical utility of acid phosphatase as a biochemical marker for the identification of C. perfringens isolated from water and other sources and its use in the differentiation of C. perfringens from ϳ95% of all currently recognized clostridial species in the genus (42).Acid phosphatases (EC 3.1.3.2) are ubiquitous and catalyze, at an acidic pH, the transfer of phosphoryl groups from phosphomonoesters to water (48). These enzymes play essential roles in the generation, acquisition, and mobilization of inorganic phosphate and critical roles in phosphoryl relay systems intimately involved in signal transduction pathways in both prokaryotes and eukaryotes. A subgroup of phosphatases has been designated nonspecific acid phosphatases (NSAPs) by Thaller and coworkers (45) and includes those bacterial polyspecific phosphatases that are secreted across the cytoplasmic membrane and exhibit optimum catalytic activity at an acidic pH (41). NSAPs are recognized as a distinct group of phosphatases and categorized into three classes (46). The class C enzyme...

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“…It has been described in literature that E. coli phosphatase activity could be lowered significantly by using an agp-negative strain like CGSC8974 and growing it in LB medium, supplemented with glucose and phosphate [13,27,28]. This effect was quantified and the specificity of E. coli phosphatases for αGlc1P and βGlc1P was evaluated (Fig.…”

Section: Production Of the Biocatalystsmentioning

confidence: 99%

Enzymatic production of β‐D‐glucose‐1‐phosphate from trehalose

Borght¹,

Desmet²,

Soetaert³

2010

Biotechnology Journal

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beta-D-Glucose-1-phosphate (beta Glc1P) is an efficient glucosyl donor for both enzymatic and chemical glycosylation reactions but is currently very costly and not available in large amounts. This article provides an efficient production method of beta Glc1P from trehalose and phosphate using the thermostable trehalose phosphorylase from Thermoanaerobacter brockii. At the process temperature of 60 C, Escherichia coli expression host cells are lysed and cell treatment prior to the reaction is, therefore, not required. In this way, the theoretical maximum yield of 26% could be easily achieved. Two different purification strategies have been compared, anion exchange chromatography or carbohydrate removal by treatment with trehalase and yeast, followed by chemical phosphate precipitation. In a next step, beta Glc1P was precipitated with ethanol but this did not induce crystallization, in contrast to what ist observed with other glycosylphosphates. After conversion of the product to its cyclohexylammonium salt, however, crystals could be readily obtained. Although both purification methods were quantitative (>99% recovery), a large amount of product (50%) was lost during crystallization. Nevertheless, a production process for crystalline beta Glc1P is now available from the cheap substrates trehalose and inorganic phosphate

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Identification of an Escherichia coli periplasmic acid phosphatase containing of a 27 kDa-polypeptide component

Cited by 19 publications

References 12 publications

Assimilation of Nicotinamide Mononucleotide Requires Periplasmic AphA Phosphatase inSalmonella enterica

Assimilation of Nicotinamide Mononucleotide Requires Periplasmic AphA Phosphatase inSalmonella enterica

Characterization of a Unique Class C Acid Phosphatase from Clostridium perfringens

Enzymatic production of β‐D‐glucose‐1‐phosphate from trehalose

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