Cloning, characterization and molecular analysis of a metalloprotease from Proteus mirabilis

Proteus mirabilis is an opportunistic pathogen that can cause urinary tract infection in human beings. The accurate and rapid identification and quantification of P. mirabilis is necessary for early treatment. In this study, a pair of specific primers according to the conserved ureR sequence of P. mirabilis was designed and novel systems which consisted of a polymerase chain reaction (PCR) and a real-time PCR to identify and quantify P. mirabilis were developed. For the qualitative identification by ordinary PCR, a 225-bp DNA product was amplified from P. mirabilis and separated on an agarose gel. The corresponding DNA product is present in three P. mirabilis strains isolated from different geographical locations, but is absent in 20 strains representing 18 different species, including the ureR homolog contained Providencia stuartii and Escherichia coli strains, the other common pathogens Klebsiella sp., Edwarsiella sp., Vibrio sp., Enterobacter sp., and Escherichia sp., and other environmental bacteria Pseudomonas sp. and Acinetobacter sp. Proteus mirabilis at concentrations higher than 1.0× 10 3 CFU ml −1 was detectable by ordinary PCR; P. mirabilis at concentrations higher than 10 CFU ml −1 was quantified by real-time PCR. The specific, sensitive and time-efficient PCR methods were demonstrated to be applicable to rapid identification and quantification of P. mirabilis.

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Quick identification and quantification of Proteus mirabilis by polymerase chain reaction (PCR) assays

Zhang

Niu

Yin

et al. 2012

Ann Microbiol

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Mirabilysin

Kerr

2013

Handbook of Proteolytic Enzymes

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Optimization of β-Glucosidase Production byAspergillus terreusStrain EMOO 6-4 Using Response Surface Methodology Under Solid-State Fermentation

El‐Naggar

Haroun

Owis

et al. 2014

Preparative Biochemistry and Biotechnology

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Forty-two morphologically different fungal strains were isolated from different soil samples and agricultural wastes and screened for β-glucosidase activity under solid-state fermentation. Eight species were chosen as the most active β-glucosidase producers and were subjected to primary morphological identification. β-Glucosidase was highly produced by Aspergillus terreus, which showed the highest activity, and was subjected to full identification using scanning electron microscopy and molecular identification. Initial screening of different variables affecting β-glucosidase production was performed using Plackett-Burman design and the variables with statistically significant effects were identified. The optimal levels of the most significant variables with positive effect and the effect of their mutual interactions on β-glucosidase production were determined using Box-Behnken design. Fifteen variables including temperature, pH, incubation time, inoculum size, moisture content, substrate concentration, NaNO3, KH2PO4, MgSO4 · 7H2O, KCl, CaCl2, yeast extract, FeSO4 · 7H2O, Tween 80, and (NH4)2SO4 were screened in 20 experimental runs. Among the 15 variables, NaNO3, KH2PO4 and Tween 80 were found as the most significant factors with positive effect on β-glucosidase production. The Box-Behnken design was used for further optimization of these selected factors for better β-glucosidase production. The maximum β-glucosidase production was 4457.162 U g(-1).

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Cloning, characterization and molecular analysis of a metalloprotease from Proteus mirabilis

Cited by 3 publications

References 34 publications

Quick identification and quantification of Proteus mirabilis by polymerase chain reaction (PCR) assays

Quick identification and quantification of Proteus mirabilis by polymerase chain reaction (PCR) assays

Mirabilysin

Optimization of β-Glucosidase Production byAspergillus terreusStrain EMOO 6-4 Using Response Surface Methodology Under Solid-State Fermentation

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