Optimization and comparative characterization of neuraminidase activities from Pseudomonas aeruginosa with Klebsiella pneumoniae,Hep-2 cell, sheep kidney and rat liver lysosome
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Abstract
Background and Objectives: The properties of neuraminidase produced by P. aeruginosa strain PAO1 during growth in a defined medium (BHI) was examined and compared with some neuraminidase features of K. pneumoniae in this investigation.
Materials and Methods: The enzyme was isolated from concentrated culture supernatants of P. aeruginosa which was used in a sensitive fluorometric assay by using 2'-(4-methylumbelliferyl) α-D-N acetylneuraminic acid as substrate.
Results: Neuraminidase production in P. aeruginosa PAO1 paralleled bacterial growth in defined medium (BHI) and was maximal in the late logarithmic phase of growth but decreased during the stationary phase, probably owing to protease production or thermal instability. Highest production of P. aeruginosa PAO1 neuraminidase was in BHI culture media. The neuraminidase of P. aeruginosa PAO1 possessed an optimum temperature of activity at 56 °C and the activity was maximal at pH 5. Heating the enzyme to 56 °C for 45 min., in the presence of bovine serum albumin destroyed 33.1% of it's activity and addition of Ca+2, EDTA and NANA also decreased activity markedly.
Conclusion: The results revealed that the highest specific activity is for p. aeruginosa PAO1.
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| Issue | Vol 2 No 1 (2010) | |
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| K. pnumoniae P. aeruginosa PAO1 Neuraminidase fluorometric assay specific activity Iran | ||
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