铜绿假单胞菌与肺炎克雷伯菌、Hep-2细胞、羊肾和大鼠肝溶酶体中神经氨酸酶活性的优化及比较特性研究

Optimization and comparative characterization of neuraminidase activities from Pseudomonas aeruginosa with Klebsiella pneumoniae, Hep-2 cell, sheep kidney and rat liver lysosome.

作者信息

Ghazaei C, Ahmadi M, Hosseini Jazani N

机构信息

Department of Microbiology, Faculty of Veterinary Medicine, University of Urmia, Urmia, Iran.

出版信息

Iran J Microbiol. 2010 Mar;2(1):30-7.

PMID:22347548

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3279765/

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.

摘要

背景与目的

本研究检测了铜绿假单胞菌PAO1菌株在限定培养基（脑心浸液培养基，BHI）中生长时产生的神经氨酸酶的特性，并与肺炎克雷伯菌的一些神经氨酸酶特性进行了比较。

材料与方法

从铜绿假单胞菌浓缩培养上清液中分离该酶，以2'-(4-甲基伞形酮基)-α-D-N-乙酰神经氨酸为底物，采用灵敏的荧光测定法。

结果

铜绿假单胞菌PAO1中神经氨酸酶的产生与细菌在限定培养基（BHI）中的生长平行，在对数生长后期达到最大，但在稳定期下降，可能是由于蛋白酶的产生或热不稳定性。铜绿假单胞菌PAO1神经氨酸酶在BHI培养基中产量最高。铜绿假单胞菌PAO1的神经氨酸酶活性最适温度为56°C，在pH 5时活性最大。在牛血清白蛋白存在下，将酶加热至56°C 45分钟，其活性丧失33.1%，添加Ca(+2)、EDTA和N-乙酰神经氨酸（NANA）也显著降低活性。